JP2002275082A - Liver function protector or improver - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a liver function protecting or improving agent, a food or drink or feed having a liver function protecting or improving action, a food or drink additive or a feed additive having a liver function protecting or improving action. And a method for screening a liver function protecting or improving agent. SOLUTION: A liver function protector or an improving agent containing a Saxifragaceae plant or an extract of the plant as an active ingredient, and a food or drink to which a Saxifragaceae plant or the extract of the plant is added. Alternatively, a feed, food or food additive or feed additive having a liver function protecting or ameliorating effect characterized by containing a plant of the family Saxifragaceae or an extract of the plant, or a substance to be screened is administered to an animal. A method for screening for a protective or ameliorating agent for liver function, which comprises evaluating a substance that decreases the GPT or GOT level in the blood of an animal that is increased by administration of lipopolysaccharide after administration of alcohol.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a liver function protecting or improving agent, a food or drink or feed having a liver function protecting or improving effect, and a food or drink additive or feed having a liver function protecting or improving effect. The present invention relates to a method for screening an additive, a liver function protecting agent or an improving agent, and a method for protecting or improving an animal liver function.

[0002]

2. Description of the Related Art The liver plays various important functions, such as controlling metabolism and storage of carbohydrates, proteins, and lipids, which are three major nutrients, and decomposing and detoxifying substances unnecessary for living organisms. It is an organ. These functions are acutely or chronically impaired by alcohol overdose, viral infection, disturbed eating habits, stress, smoking, etc., and as the disorder progresses, for example, acute hepatitis, chronic hepatitis, liver sclerosis, It causes diseases such as alcoholic fatty liver, hepatitis B virus, and liver cancer.

When hepatocytes are damaged by a virus, alcohol, or the like, aspartate aminotransferase (also referred to as glutamic acid-oxaloacetate transaminase; hereinafter abbreviated as GOT) or alanine aminotransferase (glutamic acid-pyruvate transaminase) in the cells. (Hereinafter, abbreviated as GPT)) and the like appear in the blood, so that the numerical value indicating the activity of these enzymes increases. Therefore, these GOT and GPT activities in blood are known as indices indicating liver dysfunction.

[0004] Drugs used to prevent or treat liver dysfunction include, for example, antiviral agents such as acyclovir;
Immunosuppressants (Ayumi of Medicine 171 Vol. 14 No. 957-11)
58, 1994, published in Medical and Dental Medicine), glutathione (Protein Nucleic Acid Enzyme 33, Vol. 9, No. 1625-1631, 1988)
Year, Kyoritsu Shuppan Co., Ltd.) is known. In addition, foods and drinks that are effective in protecting, enhancing, and improving liver function include:
For example, turmeric, Maria thistle, sesame lignan, oyster extract, liver extract and the like are known (FOOD Style 21,
Vol. 2, No. 12, 1998, Food Chemistry Newspaper).

[0005] The safflower family Hydrangea
macrophylla Seringe var. Thunbergii Makino) is hydrangea serrata (Hydrangea macrophylla in the neighboring species of hydrangea
It is said to have been produced by breeding of Seringe var. Acuminata). Sweet tea is known as a herbal medicine specially prepared in Japan, which is prepared by collecting and fermenting the leaves and branches of amacha and drying.

[0006] This sweet tea or sweet tea extract has been used as a raw material for producing flavoring (sweetening) drugs such as pills and oral fresheners (Japanese Pharmacopoeia, 12th Edition, D-31-33, 1991, Hirokawa Shoten) ). The sweet tea extract, which is a food additive (sweetener), is obtained by extracting water from sweet tea leaves and is in the form of a liquid or concentrated powder, and the sweetening component is phyllodulcine (existing Additives and natural fragrances, food material "Natural Products Handbook" 14th edition, Food and Science, 1998
Year).

[0007] Sweet tea has anticoccidial activity, antifungal activity,
It is known to have anti-ulcer activity, anti-allergic activity, hypercholesterolemia inhibitory activity, anti-periodontal fungus activity, antioxidant activity, etc. (2nd Medicine and Food Symposium Abstracts, p85, 1999) . Also, sweet tea extract is known to show a bile effect (Pharmaceutical Magazine, Vol. 114, No. 6, 401
413, 1994). Further, it is also known that sweet tea extract has an inhibitory effect on the lipid peroxidation reaction of liver microsomes in vitro (Natural Medicines, 49 (1) , 84-87, 1995 ) .
Year).

However, it is not known that saxifragaceous plants such as sweet tea and saxifrage have a liver function protecting or improving effect. D-galactosamine is used as a method for screening for a protective agent or an improving agent for liver function.
[Journal of Nutrition
trition), 129, 1361 (1999 )] and acetaminophen "Plant Medica (Planta Medica), 55, 417 (1989)] or carbon tetrachloride [Fundamental and Clinical pharmacophore Biology (Fundamental & Clinical Pharm
Agriculture), 3 , 183 (1989)] and the like, and using rodents with elevated blood GPT and GOT.

On the other hand, as a method for increasing GPT and GOT activities in blood, a method of administering lipopolysaccharide after administering ethanol [Gastroenology
terology), 115 , 443 (1998)]. According to this method, liver damage caused by a process similar to alcohol administration can be induced in a short period of time, but it has not been known to use this method as a screening method for a protective agent or an improving agent for liver function so far. .

[0010]

There is a need for the development of drugs that can effectively prevent or treat liver diseases, and healthy foods and drinks or animal feeds that can prevent or treat liver disorders when taken daily. An object of the present invention is to provide a liver function protector or improving agent, a food or drink or feed having a liver function protecting or improving effect, a food or drink additive or a feed additive having a liver function protecting or improving effect, a liver function. An object of the present invention is to provide a method for screening a protective agent or an improving agent, and a method for protecting or improving the liver function of an animal.

[0011]

Means for Solving the Problems The present invention provides the following (1).
To (51). (1) A liver function-protecting or improving agent comprising a plant of the family Saxifragaceae or an extract of the plant as an active ingredient. (2) The liver function-protecting or improving agent according to the above (1), wherein the plant of the family Saxifragaceae belongs to the genus Saxifraga.

(3) The agent for protecting or improving liver function according to (2), wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb. (4) The liver function-protecting or improving agent according to the above (1), wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. (5) A plant belonging to the genus Hydran
Thunbergii Makino or gea macrophylla Seringe var. Thunbergii Makino or sweet tea (Hydrangeae Dulcis Folium).

(6) The agent for protecting or improving liver function according to any of (1) to (5) above, wherein the extract of Saxifragaceae plant is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant. Agent. (7) The liver function-protecting or improving agent according to any of (1) to (6), which is orally administered.

(8) The agent for protecting or improving liver function according to any one of (1) to (7), wherein the liver function is a function affected by alcohol.

(9) A food or drink to which a plant of the family Saxifragaceae or an extract of the plant is added. (10) The food or drink according to (9), wherein the food or drink is used for protecting or improving liver function. (11) The food or drink according to (10), wherein liver function is affected by alcohol.

(12) The food or drink as described in any of (9) to (11) above, wherein the Saxifragaceae plant is a plant belonging to the genus Saxifraga. (13) The plant according to (12), wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.
Food and drink as described. (14) The food or drink according to any of (9) to (11), wherein the saxifragaceae plant is a plant belonging to the genus Hydrangea.

(15) Hydrangea macrophylla Seringe var. Thunbergi
i. Makino) or sweet tea ( Hydrangeae Dulcis Folium). (16) The food or drink product according to any of (9) to (15), wherein the Saxifragaceae plant extract is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant.

(17) A feed obtained by adding a Saxifragaceae plant or an extract of the plant. (18) The feed according to the above (17), wherein the feed is used for protecting or improving liver function. (19) The feed according to (18), wherein the liver function is a function affected by alcohol.

(20) The feed according to any one of (17) to (19), wherein the Saxifragaceae plant is a plant belonging to the genus Saxifraga. (21) The plant according to the above (20), wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.
Feed as described. (22) The feed according to any of (17) to (19), wherein the saxifragaceae plant is a plant belonging to the genus Hydrangea.

(23) Hydrangea macrophylla Seringe var. Thunbergi
i Makino) or sweet tea (Hydrangeae Dulcis Folium). (24) The feed according to any one of the above (17) to (23), wherein the Saxifragaceae plant extract is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant.

(25) An additive for foods and drinks having a hepatic function-protecting or ameliorating action, characterized by containing a plant of the family Saxifragaceae or an extract of the plant. (26) The additive for food and drink according to (25), wherein the liver function is a function affected by alcohol. (27) The additive for food and drink according to the above (25) or (26), wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga.

(28) A plant belonging to the genus Saxifraga,
The additive for foods and drinks according to the above (27), which is Saxifraga stolonifera Meerb. (29) The additive for food or drink according to the above (25) or (26), wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. (30) plant belonging to the hydrangea is, armature (Hydr
(2) which is angea macrophylla Seringe var. Thunbergii Makino or sweet tea (Hydrangeae Dulcis Folium)
9) The additive for food and drink according to the above.

(31) The extract of Saxifragaceae plant is
The food or beverage additive according to any one of the above (25) to (30), which is an alcohol extract of an aqueous medium extract residue of a Saxifragaceae plant. (32) An additive for feed having a hepatic function-protecting or improving action, comprising a plant of the family Saxifragaceae or an extract of the plant.

(33) The additive for feed according to (32), wherein the liver function is a function affected by alcohol. (34) The additive according to (32) or (33), wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga. Feed additives. (35) The plant according to (34), wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.
The feed additive as described in the above.

(36) The feed additive according to the above (32) or (33), wherein the saxifragaceae plant is a plant belonging to the genus Hydrangea. (37) plant belonging to the hydrangea is, armature (Hydr
(3) which is angea macrophylla Seringe var. Thunbergii Makino or sweet tea (Hydrangeae Dulcis Folium)
6) The additive for feed according to the above.

(38) The extract of Saxifragaceae plant is
The feed additive according to any one of the above (32) to (37), which is an alcohol extract of an aqueous medium extract residue of a Saxifragaceae plant.

(39) administering a substance to be screened to an animal to evaluate a substance which decreases the GPT or GOT level in the blood of the animal which is increased by administration of lipopolysaccharide after administration of alcohol. Screening method for an agent for protecting or improving liver function.

(40) The above (3), wherein the animal is a mammal.
9) The method described in the above. (41) The method according to any of (39) or (40) above, wherein the lipopolysaccharide is derived from a microorganism belonging to the group of intestinal bacteria. (42) The method according to any of (39) to (41), wherein the liver function is a function affected by alcohol.

(43) The liver function-protecting or improving agent according to any of (1) to (8) or the (1)
7) A method for protecting or improving liver function of an animal, comprising feeding the animal with any of the feeds according to any one of 7) to (24). (44) The above-mentioned (4), wherein the animal is livestock, poultry, or cultured fish.
The method according to 3).

(45) A food, drink or feed for protecting or improving liver function, which comprises a plant of the family Saxifragaceae or an extract of the plant as an active ingredient.

(46) The food or drink or feed for protecting or improving liver function according to the above (45), wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga. (47) The food or drink or feed for protecting or improving hepatic function according to (46), wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.

(48) The food or drink or feed for protecting or improving liver function according to the above (45), wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. (49) plant belonging to the hydrangea is, armature (Hydr
Thunbergii Makino) or sweet tea (Hydrangeae Dulcis Folium) (4) which is an angea macrophylla Seringe var.
8) A food or drink or feed for protecting or improving liver function according to the above.

(50) The extract of Saxifragaceae plant is
The food or drink or feed for protecting or improving hepatic function according to any of (45) to (49), which is an alcohol extract of an aqueous medium extract residue of a Saxifragaceae plant. (51) The food or drink or feed for protecting or improving liver function according to any of (45) to (50), wherein the liver function is a function affected by alcohol.

[0034]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, protection of liver function includes an action of protecting liver function from various disorders and an action of preventing liver function from disorders. Improving liver function is an action that restores or heals the function of the affected liver,
Includes the action of improving or enhancing the function of the liver.

In the present invention, the Saxifragaceae plant means a plant belonging to the scientific name Saxifragaceae [Makino Shin Nihon Botanical Book (40th edition, June 10, 1984)
Sun, Hokuryukan), Primary Color Makino Plant Encyclopedia (Sequel) (First Edition Showa)
May 10, 1958, Hokutakakan)]. Saxifragaceae plants include, for example, genus Astilbe and cornflower ( R
odgersia) genus, saxifrage (Saxifraga) genus, rock saxifrage (Tanakaea) genus, Aserifiramu (Aceriphyllum)
Genus, Alasugusa ( Boykinia ), Cats ( Chryso )
splenium) genus, Sudayakushu (Tiarell) genus, Charumerusou (Mitella) genus Parnassia palustris (Parnassia) genus,
Syringa ( Philadelphus ) genus, Deutzia ( Deutzia )
Genus, baika ( Platycrater ), hydrangea ( Hyd )
rangea) genus, Iwagarami (Schizophragma) genus, Kusaajisai (Cardiandra) genus, Ginbaisou (Deinanthe)
Genus, genus Itea, genus Currant ( Ribes ), genus Kirengeshoma, and plant varieties bred from these plants, and the like. Plant varieties bred so that the improving effect is enhanced are preferred.

[0036] Examples of the species belonging to the genus Chidakesashi, for example Chidakesashi (Astilbe microphylla Knoll), Akashouma (Astilbe thunbergii Miq.), Triacylate Cimicifuga (A
stilbe odontophylla Miq.), millet Mori Cimicifuga (Asti
lbe japonica Miq.), Aedes zebra ( Astilbe sim)
plicifolia Makino). As a species belonging to the genus Cornflower , for example, Cornflower ( Rodgersia)
podophylla A. Gray).

Examples of species belonging to the genus Saxifraga include, for example, Saxifraga stolonifera Meerb., Saxifraga nipponica Makino, Saxifola for Saxifraga fortunei Hook., Sajifraga fortunei Hook.
fraga cortusaefolia Sieb.et Zucc.), Fukiyukinosita ( Saxifraga japonica Boiss.), Black moss ( Sax
ifraga fusca Maxim), Maple biloba ( Saxifr)
aga sendaica Maxim.var. laciniata Nakai), spider magsa ( Saxifraga merkii Fisch. var. idsuroei Eng)
l.), Kumaguma snow blossom ( Saxifraga laciniata Nakai
et Takeda), Gypsophila ( Saxifraga bronchialalis )
L.), Saxifraga cernua L., and Saxifraga sachalinensis Fr. Schm., And the like, with Saxifraga being preferred.

[0038] Examples of the species belonging to Iva Saxifraga, for example Iva saxifrage (Tanakaea rad icans Franch. Et S
av.) and the like.

Examples of the species belonging to the genus Aseriphyllum include, for example, Asperilla ( Aceriphyllum rossii Engler).

Examples of the species belonging to the genus Arasigusa include, for example, alasigsa ( Boykinia lycoctonifolia Engl.) And Yallacea ( Boykinia tellimoides Engl. Et Irmsch.).

Examples of the species belonging to the genus Catharsis include cathars ( Chrysosplenium grayanum Maxim.),
Red cat flies ( Chrysosplenium stamineum Franc
h), lynx ( Chrysosplenium japonicum Ma)
kino), Periwinkle ( Chrysosplenium flagellif)
erum Fr.Schm.), Lynx ( Chrysospleniu )
m macrostemon Maxim.), Scutellaria ( Chryso )
splenium sphaerospermum Maxim.).

[0042] Examples of the species belonging to the genus Sudayakushu, for example T. polyphylla (Tiarella polyphylla Don), and the like. The species belonging to mitella, e.g. Charumerusou (Mitella japonica Miq.), Kocharumerusou (Mitella pauciflora Rosend.) And the like.

Species belonging to the genus Hornets include, for example, hornets ( Parnassia palustris L.), hornets ( Parnassia alpicola Makino), and whisker ( Parnassia foliosa Hook.f.et Thoms.var. Nummula)
ria Nakai).

The species belonging to the genus Syringa include, for example, Syringa ( Philadelphus satsumi Sieb.) And the like.

Examples of the species belonging to the genus Aspergillus include, for example, aspergillus ( Deutzia crenata Sieb. Et Zucc.), Maruberagi ( Deutzia sieboldiana Maxim.), And Astragalus ( Deutzi ).
a gracilis Sieb.et. Zucc.) and Vladivostalis ( Deutz )
ia maximowicziana Makino), Umetsugi ( Deutzia un
iflora Shirai), Aomori azalea ( Deutzia gracilis)
Var. Nagurai Makino) and the like.

Examples of the species belonging to the genus Baikama include Baikama ( Platycraterserrata Makino) and the like. Examples of species belonging to the genus Hydrangea include ginkgo hydrangea ( Hydrangea macrophylla Seringe) and hydrangea ( Hydrangea macrophylla Seringe var. Otaksa).
Makino) and red garlic ( Hydrangea macrophylla Seringe)
subsp. serrata Makino var. japonica Makino), Hydrangea macrophylla Seringe var. acumi
nata), amateur ( Hydrangea macrophylla Seringe va)
r. Thunbergii Makino), sweet tea (Hydrangeae Dulcis Fo)
lium), gakuutsugi ( Hydrangea scandens Seringe),
Cod rhinoceros ( Hydrangea hirta Sieb. Et Zucc.), Tama hydrangea ( Hydrangea involucrata Sieb.), Hydrangea hydrangea ( Hydrangea sikokiana Maxim.), Norris cypress ( Hydrangea paniculata Sieb.), Tsurumari ( Hydran )
gea petiolaris Sieb. et Zucc.), Hydrangea ( Hyd )
rangea macrophylla Seringe subsp.serrata Makino v
ar. amoena Makino), Hydrangea macro
phylla Seringe subsp.serrata Makino var.angustat
a Makino) and the like, and amcha or sweet tea is preferred, and sweet tea is particularly preferred.

Examples of the species belonging to the genus Genus include, for example, Schizophragma hydrangeoides Sieb. Et Z
ucc.) and the like.

As a species belonging to the genus Rhododendron, for example, hydrangea ( Cardiandra alternifolia Sieb. Et Z
ucc.) and the like.

The species belonging to the genus Ginseng include, for example, ginseng ( Deinanthe bifida Maxim.) And the like.

Examples of species belonging to the genus Zuina include, for example, Zina ( Itea japonica Oliver).

Examples of the species belonging to the genus gooseberry include, for example, gooseberry ( Ribes sinanense F. Maekawa), Atlantic gooseberry ( Ribes grossularia L.), and Ezo gooseberry ( Ribes latifo ).
lium Jancz.), red currant ( Ribes rubrum L.), red currant ( Ribes japonicum Maxim.), horse currant ( Ribes sachalinense Nakai), and raspberry ( Ribesf )
asciculatum Sieb. et Zucc.), Zarikomi ( Ribes alpi )
num L. var. japonicumMaxim.), Rib
es ambiguum Maxim.).

[0052] As the species belonging to the genus Kirengeshoma,
For example, Kirengeshoma palmata Yatab
e) and the like.

The plant of the present invention includes, for example, leaves, flowers, branches, stems, and the like of wild plants, plants obtained by cultivation, or plants obtained by culture such as tissue culture.
Examples include fruits, roots, seeds, cultured cells or organs, callus, and the like, and various processed products obtained by treating these as such or physically, chemically, or biologically.

Examples of the physical / chemical treatment method include a drying treatment such as sun drying, air drying and freeze drying, and a pulverization treatment with a blender, a homogenizer, a ball mill, and the like. , Dried, freeze-dried, pulverized, etc. Examples of the biological treatment method include a fermentation method, and examples of the biological treatment product include a fermentation treatment product.

Examples of the plant extract include extracts obtained from the above-mentioned plants by various extraction methods. Examples of the extraction method include various solvent extraction and supercritical fluid extraction. Extraction method, sedimentation separation, cake filtration, clarification filtration, centrifugal filtration, centrifugal sedimentation, compression separation, various solid-liquid separation methods such as filter press, various concentration methods, various drying methods, formulation methods such as granulation or powdering, You may process by various purification methods etc.

Examples of the purification method include a solvent fractionation method, a column chromatography method, a recrystallization method and the like. In particular,
A column chromatography method using various carriers such as Diaion HP-20 (manufactured by Mitsubishi Chemical Corporation) and Sephadex LH-20 (manufactured by Pharmacia) is preferred. Concentration and drying methods include freeze drying, air drying, hot air drying,
Ventilation drying, blast drying, spray drying, vacuum drying, solar drying,
Drying methods such as vacuum drying, fluidized bed drying, foam layer drying, and film drying methods such as drum dryers, ultrasonic drying and electromagnetic wave drying, preferably spray drying methods and freeze drying methods are mentioned.

In the extraction and the treatment of the extract, for example, an antioxidant or a preservative can be added. As the solvent used for the solvent extraction, any solvent can be used as long as it can extract a substance exhibiting a liver function-protecting action or a liver function-improving action of the present invention. Aqueous media such as liquids, monohydric alcohols such as methanol, ethanol, propanol and butanol, polyhydric alcohols such as propylene glycol and glycerol, hexane, toluene, petroleum ether, benzene, ethyl acetate, chloroform, dichloromethane, 1,1,2
-Organic solvents such as trichloroethene, dimethylsulfoxide, and acetone, and the like, and aqueous media and alcohols are preferred.

Examples of the buffer include a phosphate buffer, a citrate buffer and the like. Examples of the inorganic salt in the aqueous inorganic salt solution include sodium chloride, potassium chloride, calcium chloride and the like. As the alcohol, a monohydric alcohol is preferable, and as the monohydric alcohol, ethanol is preferable.

These solvents can be used alone or in combination of two or more. As the mixed solvent, hydrous alcohol is preferable, hydrous monohydric alcohol is more preferable, and hydrous ethanol is particularly preferable. The water content is 70%
Or less, more preferably 40% or less. As the solvent, carbon dioxide that has been made into a supercritical fluid can also be used.

The extraction is performed using, for example, 0.1 part by weight to 10000 parts by weight, preferably 1 part by weight to 100 parts by weight of a solvent per 1 part by weight of a plant. The extraction temperature is not particularly limited, but is preferably 0 ° C to 100 ° C, more preferably 20 ° C to 90 ° C. The extraction time is not particularly limited, but is from 1 minute to 1 minute.
Weeks are preferred and 30 minutes to 1 day are more preferred. In particular, as a method for extracting from Saxifragaceae plants, the aforementioned Saxifraga plants, the physical and chemical treatments of the aforementioned Saxifraga plants, the above-described biologically treated products are extracted with an aqueous medium using alcohol or hydrous alcohol. The extraction method is preferred.

The aqueous medium is not particularly limited.
Water, pure water and deionized water are preferred. The extraction temperature when extracting with an aqueous medium and alcohol or hydrous alcohol is not particularly limited, but is preferably 0 ° C to 100 ° C,
0 ° C to 90 ° C is more preferred. The extraction time is not particularly limited, but is preferably 1 minute to 1 week, more preferably 30 minutes to 1 day. It is preferable to use a plant that has been subjected to a drying treatment or a fermentation treatment as a saxifrage plant.

The equipment used for the extraction is not particularly limited, but includes a container, a stirrer, a reflux condenser, a Soxhlet extractor, a homogenizer, a shaker, an ultrasonic generator, etc., which are devised for efficient extraction. can give. The liver function-protecting agent or ameliorating agent of the present invention contains a Saxifragaceae plant prepared by the above-described method or an extract thereof, and if necessary, one or more pharmacologically acceptable carriers, and further, May contain active ingredients for other treatments.

The present pharmaceutical preparation is produced by mixing a Saxifragaceae plant or the extract thereof with a carrier if necessary, and by any method well known in the technical field of pharmaceuticals. The dosage form of the preparation is desirably the one most effective in treatment, and may be oral administration or parenteral administration such as intravenous, intraperitoneal or subcutaneous administration. Of these, oral administration is preferred.

The dosage forms to be administered include tablets, powders, granules, pills, suspensions, emulsions, diluents, capsules, syrups, injections, solutions, elixirs, extracts, tinctures, and liquid extracts. Agents and the like. For example, extracts, tinctures, fluid extracts, etc. suitable for oral administration are prepared by directly extracting or concentrating an extract of a plant of the family Saxifragaceae, such as water, ethanol, or a mixture of water and ethanol. can do.

Liquid preparations suitable for oral administration, such as syrups, include water, sugars such as sucrose, sorbitol, fructose, glycols such as polyethylene glycol and propylene glycol, oils such as sesame oil, olive oil and soybean oil. , A preservative such as p-hydroxybenzoic acid esters, a paraoxybenzoic acid derivative such as methyl paraoxybenzoate, a preservative such as sodium benzoate, and a carrier such as flavors such as strawberry flavor and peppermint. .

Suitable for oral administration, for example, tablets, powders and granules include lactose, sucrose, glucose, sucrose,
Mannitol, saccharides such as sorbitol, potato, wheat, starch such as corn, calcium carbonate, calcium sulfate, sodium hydrogen carbonate, inorganic substances such as sodium chloride, crystalline cellulose, excipients such as licorice powder, plant powder such as gentian powder, Disintegrants such as starch, agar, gelatin powder, crystalline cellulose, carmellose sodium, carmellose calcium, calcium carbonate, sodium bicarbonate, sodium alginate, magnesium stearate, talc, hydrogenated vegetable oil, macrogol, silicone oil, etc. Binders, polyvinyl alcohol, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, binders such as carmellose, gelatin, starch paste, surfactants such as fatty acid esters, plasticizers such as glycerin, etc. It can be prepared using.

For example, injections suitable for parenteral administration comprise preferably sterile aqueous solutions containing the active compound which are isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared using a carrier comprising a salt solution, a glucose solution or a mixture of saline and a glucose solution. In these parenteral preparations, the above-mentioned preservatives, preservatives, surfactants and the like can be used.

The dose and the number of administrations of the agent for protecting or improving liver function of the present invention are determined depending on the administration form, age, body weight,
Depending on the nature or severity of the condition to be treated,
There is no particular limitation, and in the case of oral administration, 0.01 mg to 50 g, preferably 0.05 mg to 10 g, once a day, as a dry weight of a saxifragaceous plant or an extract of the plant extract per adult per day. Or several times.
In the case of parenteral administration such as intravenous administration, 0.001 mg to 50 g, preferably 0.01 mg to 10 g, per day, as a dry weight of a saxifragaceous plant or an extract of the plant extract per adult per day. One or several doses. When administered to an animal, it varies depending on the age, type, nature or severity of the condition of the animal, but is not particularly limited, and is preferably 0.1 μg to 10 g, preferably 1 μg to 1 g per day per 1 kg of body weight. One or several doses. In the case of parenteral administration such as intravenous administration, the body weight is 1 kg1.
0.01 μg to 10 g, preferably 1 μg to 1 g / time
g once or several times daily. However, the dose and the number of doses vary depending on the various conditions described above.

The food or drink or feed having a hepatic function-protecting or ameliorating effect obtained by adding a plant of the family Saxifragaceae or an extract of the plant, the plant of the present invention or the plant of the present invention may be used as a food or drink or feed. Add the extract,
Examples include those produced by a general method of producing foods and drinks or feeds. The food or drink or feed may be a general food or drink or feed processed by, for example, a molding and granulating method. Molding and granulation methods include fluidized bed granulation, stirring granulation, extrusion granulation, tumbling granulation, airflow granulation, compression molding granulation, crushing granulation, spray granulation, spray granulation, etc. Granulation method, pan coating, fluidized bed coating, dry coating,
Such as coating method, puff drying, excess steam method,
Examples thereof include expansion methods such as a foam mat method and a microwave heating method, and extrusion methods such as an extrusion granulator and an extruder.

As foods or drinks or feeds for protecting or improving hepatic function characterized by containing a plant of the family Saxifragaceae or an extract of the plant as an active ingredient, a plant of the family Saxifragaceae or an extract of the plant as it is It can be used as a product or feed. Further, a food or drink or feed obtained by adding a Saxifragaceae plant or an extract of the plant to a food or drink or feed or a raw material thereof can be used.

There are no particular restrictions on the food, drink, feed or raw materials to which the plant of the family Saxifragaceae or the extract of the plant is added, and there is no particular limitation on the plant of the family Saxifragaceae or the extract containing the plant extract. The plant may not substantially contain a Saxifragaceae plant or the plant extract. By adding a Saxifragaceae plant or an extract thereof to a food or drink or a feed containing the Saxifragaceae plant or the extract of the plant, a hepatic function protecting or improving effect of the food or drink or the feed Can be increased.

The amount of the Saxifragaceae plant or the plant extract of the present invention to be added to foods and drinks or feeds is such that the foods or drinks or feeds can be brought to a concentration at which the liver function is protected or improved. Is not particularly limited as long as it is 0.001 to 100%, preferably 0.01 to 100%, more preferably 0.1 to 100%, as dry weight of Saxifragaceae plant or dry weight of the plant extract. 1
It is added so as to contain 00%.

Specific foods and drinks to which the Saxifragaceae plant or the plant extract is added include, for example, juices, soft drinks, soups, teas to which the Saxifragaceae plant or the plant extract is added. Dairy products such as dairy products, lactic acid beverages, fermented milk, frozen desserts, butter, cheese, yogurt, processed milk, skim milk powder, livestock meat products such as ham, sausage, hamburgers, fish meat refining products, egg rolls, egg tofu, etc. ,
Examples include cookies, jellies, snacks, confectionery such as chewing gum, breads, noodles, pickles, smoked products, dried fish, boiled food, seasonings, and the like.

Examples of the form of food and drink include powdered foods,
Sheet food, bottled food, canned food, retort food,
Examples include capsule foods, tablet foods, liquid foods, drinks and the like. The food and drink of the present invention are used as healthy food and drink and functional food and drink for protecting or improving liver function.

When a food or drink having a hepatic function-protecting or ameliorating effect containing the Saxifragaceae plant of the present invention or the plant extract as an active ingredient is taken, the amount of intake is not particularly limited. On the other hand, it is 0.1 g to 50 g, preferably 0.5 to 10 g, as dry weight of Saxifragaceae plant or extract of the plant extract per day. This intake is taken for one day to one year, preferably for two weeks to three months. However, this intake amount is only a guide, and can be appropriately adjusted to a suitable range according to the degree of the symptom, age, weight, and the like of the user.

Examples of the feed of the present invention include a feed obtained by adding a Saxifragaceae plant or an extract of the plant to a feed raw material. As the feed of the present invention, mammals, birds, reptiles, amphibians, fish and other animals may be any feed as long as it has a liver function protecting or improving effect, for example, dogs, cats, rats, etc. Pet feed,
Examples include feed for livestock such as cows and pigs, feed for poultry such as chickens and turkeys, and feed for cultured fish such as Thailand and hamachi.

The feed of the present invention can be prepared by appropriately mixing a Saxifragaceae plant or an extract of the plant with feed materials. Examples of feed raw materials include cereals, rice bran, vegetable oil cake, animal feed raw materials, other feed raw materials, purified products, and the like. Grains include, for example, milo, wheat, barley, oats, rye, brown rice, buckwheat, foam, cane, hee,
Corn and soybeans.

Examples of the bran include rice bran, defatted rice bran, bran, ground powder, wheat germ, wheat bran, pellets, corn bran, corn germ and the like. Vegetable oil cakes include, for example, soybean oil cake, kinako, linseed oil cake, cottonseed oil cake, peanut oil cake, safflower oil cake, palm oil cake, palm oil cake, sesame oil cake, sunflower oil cake, and rapeseed oil cake , Kapok oil cake, mustard oil cake and the like.

Examples of animal feed materials include fish meal (north sea meal, imported meal, whole meal, coastal meal),
Fish soluble, meat meal, meat-and-bone meal, blood meal, decomposed hair, bone meal, livestock processing by-product, feather meal, silkworm, skim milk powder, casein, dried whey, and the like. Other feed materials include plant foliage (alfalfa, hay cube, alfalfa leaf meal, false acacia powder, etc.), corn processing by-products (corn gluten, meal, corn gluten feed, corn staple liquor, etc.), and processed starch (starch) Etc.), sugar, fermented industrial products (yeast, beer ground, malt root, alcohol grounds, soy ground ground, etc.), agricultural production by-products (citrus processed grounds, tofu grounds, coffee grounds, cocoa grounds, etc.), others (cassava, broad beans, Guamir, seaweed, krill,
Spirulina, chlorella, minerals, etc.).

The purified products include proteins (casein,
Albumin, etc.), amino acids, carbohydrates (starch, cellulose, sucrose, glucose, etc.), minerals, vitamins and the like. When the animal is ingested with a feed containing a saxifrage plant or the extract according to the present invention or an extract containing the extract as an active ingredient and having a hepatic function-protecting action or a hepatic function-improving action, the amount of intake is not particularly limited. 1kg
Per day, as dry weight of Saxifragaceae plant or dry weight of the plant extract per day, 0.1 mg to 50 g,
Preferably it is 0.5 mg to 10 g. This intake is 1
Continue taking for 1 day to 1 year, preferably 2 weeks to 3 months.
However, this intake amount is only a guide and can be appropriately adjusted to a suitable range according to the kind, age, body weight, etc. of the animal.

An additive for foods and drinks or an additive for feed having a hepatic function-protecting or ameliorating effect, characterized by containing a plant of the family Saxifragaceae or an extract of the plant as an active ingredient, is prepared by the aforementioned method. An additive which contains the plant of the family Saxifragaceae or an extract of the plant as an active ingredient, and is generally used as needed in foods and drinks or feeds, for example, a food additive labeling handbook (Japanese Food Additives Association, January 1997 Sweeteners described on March 6),
Colorants, preservatives, thickeners, antioxidants, color formers, bleaches, fungicides, gum bases, bitters, enzymes, brighteners,
Additives such as acidulants, seasonings, emulsifiers, enhancers, manufacturing agents, spices and spice extracts may be added. Further, the carriers exemplified in the above-mentioned pharmaceutical preparations may be added.

Examples of sweeteners include aspartame, licorice, stevia, xylose, lakanca and the like. Examples of the coloring agent include carotenoids, turmeric pigments, flavonoids, caramel pigments, turmeric pigments, spirulina pigments, chlorophyll, purple potato pigments, purple potato pigments, perilla pigments, and blueberry pigments.

Examples of the preservatives include sodium sulfite, benzoic acids, udo extract, sesame extract, radish extract, sorbic acids, propionic acids and the like. Examples of the thickening stabilizer include gums such as gum arabic and xanthan gum, alginic acids, chitin, chitosan, kidachialoe extract, guar gum, hydroxypropylcellulose, sodium caseinate, corn starch, carboxymethylcellulose, gelatin, agar, dextrin, methylcellulose, Examples thereof include polyvinyl alcohol, microfibrous cellulose, microcrystalline cellulose, seaweed cellulose, sodium polyacrylate, sodium polyphosphate, carrageenan, yeast cell wall, konjac extract, nata de coco, mannan and the like.

Antioxidants include vitamin C, sodium ethylenediaminetetraacetate, calcium ethylenediaminetetraacetate, erythorbic acid, oryzanol, catechin, quercetin, clove extract, enzyme-treated rutin, apple extract, sesame oil extract, dibutylhydroxytoluene , Fennel extract, horseradish extract, seri extract, tea extract, tempeh extract, dokudami extract, tocotrienol, tocopherols, rapeseed oil extract,
Green coffee bean extract, sunflower seeds, ferulic acid, butylhydroxyanisole, blueberry leaf extract, propolis extract, Hego ginkgo extract, hesperetin,
Examples include pepper extract, balsam extract, gallic acid, bayberry extract, eucalyptus extract, and rosemary extract.

The coloring agent includes sodium nitrite and the like, and the bleaching agent includes sodium sulfite and the like. Examples of fungicides include orthophenylphenol. Examples of the gum base include methyl acetyl ricinoleate, urushi wax, ester gum, eremi resin, aucucumber wax, ozokerite, opopanax resin, kauri gum, carnauba wax, guaiac resin, gutta kachu, gutta hankan, gutta percha, glycerin fatty acid eslu, gay wax, copaoba balsam, copal. Resin, rubber, rice bran wax, sugar cane wax, shellac, gelton, sucrose fatty acid ester, sorba, sorbitan fatty acid ester, talc, calcium carbonate, dammar resin, chicle, tilte, tunoe, low molecular rubber, paraffin wax, fir balsam, propylene glycol fatty acid Ester, powdered pulp, powdered peach, jojoba wax, polyisobutylene, polybutene, microcrystal wax, mastic, Tsu Saran Dubai chocolate, beeswax, such as calcium phosphate, and the like.

As the bittering agent, isoalpha-bittering acid,
Caffeine, Kawatake mushroom extract, Kina extract, Yellowfin extract, Gentian extract, Spice extract, Enzyme-treated naringin, Jamaica cassia extract, Theobromine, Naringin, Nigaki extract, Scutellaria extract, Hiokikoshi extract, Himematsutake extract , Borampet, methylthioadenosine, litchi extract, olive tea, daidai extract, hop extract, mugwort extract, and the like.

Examples of the enzyme or enzyme source include amylase, trypsin, rennet, lactic acid bacteria and the like. Examples of the brightener include shiro wax, mokurou and the like. Acidulants include adipic acid, itaconic acid, citric acids, succinic acids, sodium acetate, tartaric acids, carbon dioxide, lactic acid, phytic acid, fumaric acid, malic acid, phosphoric acid, and the like.

Examples of seasonings include amino acids such as asparagine, aspartic acid, glutamic acid, glutamine, alanine, isoleucine, glycine, serine, cystine, tyrosine, leucine, proline, sodium inosinate,
Nucleic acids such as sodium uridylate, sodium guanylate, sodium cytidylate, calcium ribonucleotide and sodium ribonucleotide, organic acids such as citric acid and succinic acid, potassium chloride, low-sodium salt solution in salt water, lake water, potassium salt in crude seawater, and whey salt , Tripotassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, trisodium phosphate, chlorella extract and the like.

Examples of enhancers include zinc salts, vitamin C, various amino acids, 5-adenylic acid, iron chloride, hesperidin,
Various calcined calcium, various uncalcined calcium, dibenzoylthiamine, calcium hydroxide, calcium carbonate, thiamine hydrochloride, Dunaliella carotene, tocopherol, nicotinic acid, carrot carotene, palm oil carotene, calcium pantothenate, vitamin A, hydroxyproline, Examples include calcium dihydrogen pyrophosphate, ferrous pyrophosphate, ferric pyrophosphate, ferritin, heme iron, menaquinone, folic acid, riboflavin and the like.

Examples of manufacturing agents include processing aids such as acetone and ion exchange resins, fig leaf extract, rice straw ash extract, kaolin, glycerin fatty acid ester, mulberry extract, bone ash, perilla extract, ginger extract, various Tannin, phafia pigment, grape seed extract, ethanol and the like. In addition, these various additives can also be used by adding to the above-mentioned liver function protecting agent or improving agent, and a food or drink or feed having a liver function protecting or improving effect.

The liver function in the present invention means all functions of the liver and is not particularly limited. Specific liver functions include, for example, blood storage (adjustment of circulating volume), processing of hemoglobin (processing and excretion of hemoglobin), generation of bile,
Functions in blood and circulation, such as enterohepatic circulation of bile pigments, synthesis of plasma proteins (acute phase proteins, albumin, blood clotting factors, steroid binding proteins, other hormone binding proteins, etc.), nutrients and vitamins (glucose and other sugars) Metabolic functions of nutrients such as metabolism of amino acids, lipids or fatty acids, cholesterol, lipoproteins, fat-soluble vitamins, water-soluble vitamins, etc., various substances (toxins,
Detoxification or decomposition functions such as inactivation of steroids such as estrogen and androsterone, other hormones, etc., and immune functions [“Physiological Perspective”, 19th edition of the original text
March 31, 2), "New Clinical Nutrition" Revised 3rd Edition
(May 20, 2000)]. All of these functions are impaired by alcohol.

Next, the substance to be screened is administered to the animal, so that GPT or G in the blood of the animal which is increased by administration of lipopolysaccharide after administration of alcohol is administered.
A method for screening a liver function protecting or improving agent, which is characterized by selecting a substance that reduces the OT value, will be described. As an animal used for screening, after administering alcohol, administering lipopolysaccharide,
There is no particular limitation as long as the animal has an elevated GPT or GOT level in the blood. For example, mice, rats, rabbits, dogs,
Mammals such as cats.

The substances to be screened include feed administration,
It can be administered to animals by oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, etc., but feed administration or oral administration is preferred. The substance to be screened may be administered to the animals freely or at regular intervals. For example, the substance to be screened is mixed with the feed, or separately from the feed, for 1 hour to 360 days, preferably 1 day to 2 days.
The administration is performed once a day to three times a day, more preferably for 3 days to 15 days.

The alcohol can be administered to the animal by, for example, oral administration or intravenous administration, but oral administration is preferred. The alcohol may be administered either before, in parallel with, or after administration of the substance to be screened to the animal, but preferably after administration of the substance to be screened. The amount of alcohol to be administered is 2 to 10 g / kg, preferably 3 to 5 g / kg, based on the body weight of the animal. It is preferable to use a probe or the like for administration. The administration of alcohol is, for example, 1 to 3
This can be performed 60 times, preferably 1 to 10 times. The period can be raised, for example, for 1 hour to 360 days, preferably for 1 day to 2 months, and more preferably for 3 days to 15 days. After administration of alcohol, 1 to 24 hours, preferably 3 to 9 hours, particularly preferably 6 hours, the lipopolysaccharide is added to the animal in an amount of 0.15 to 15
mg / kg, preferably 3 to 6 mg / kg, is injected intravenously, and the GPT or GOT value in the blood is quantified 12 to 36 hours, preferably 20 to 28 hours. When the substance to be screened is not given, the GP in the blood of an animal is increased by the administration of lipopolysaccharide after the administration of alcohol.
By comparing with the T or GOT value and selecting a substance that reduces the GPT or GOT value when the substance to be screened is given, screening for an agent for protecting or improving liver function can be performed.

Examples of the lipopolysaccharide include those extracted from gram-negative bacteria by a known method. Examples of Gram-negative bacteria include photosynthetic bacteria such as Rhodopseudomonas, Pseudomonas bacteria, Escherichia,
Intestinal bacteria such as Salmonella; chemolithotrophic bacteria such as Nitrobacter; Thiobacillus; methanogens such as Neisseria; Among these lipopolysaccharides, those derived from microorganisms belonging to the group of intestinal bacteria are preferred, and those derived from microorganisms belonging to the genus Escherichia are particularly preferred.

According to this screening method, a preventive or therapeutic agent for alcoholic liver injury can be suitably screened. As a method for quantifying GPT and GOT values, GP
There is no particular limitation as long as the T or GOT value can be determined. GP
The quantification of the T value can be performed, for example, by quantifying pyruvate generated from 2-oxoglutamic acid and alanine. Pyruvic acid can be quantified by, for example, coexistence of lactate dehydrogenase and NADH and quantifying the amount of decrease in absorbance at 340 nm accompanying the reduction of pyruvate.

The GOT value can be determined, for example, by determining oxaloacetic acid generated from aspartic acid and 2-oxoglutaric acid. Oxaloacetic acid can be quantified by, for example, coexistence of malic acid dehydrogener and NADH and quantifying the decrease in absorbance at 340 nm accompanying the reduction of oxaloacetic acid. In the present invention, by selecting a substance that significantly reduces the GPT or GOT value in the blood of an animal that is increased by administration of lipopolysaccharide after administration of alcohol, which is observed when the substance to be screened is not administered, An agent for protecting / enhancing / improving liver function or an agent for preventing or treating alcoholic liver injury can be selected.

Hereinafter, the present invention will be described in detail with reference to examples.
However, the following examples do not limit the present invention.

The following substances were used in the following examples. Sweet tea powder (manufactured by Shihei Shoten), Sakinoshita powder (manufactured by Shihei Shoten), D-galactosamine (manufactured by Wako Pure Chemical Industries), collagenase (manufactured by Sigma), acetaminophen (manufactured by Wako Pure Chemical Industries), penicillin (Manufactured by Gibco), streptomycin (manufactured by Gibco), DMS
O (manufactured by Wako Pure Chemical Industries), insulin (manufactured by Wako Pure Chemical Industries), dexamethasone (manufactured by Wako Pure Chemical Industries), TNF
-Α (manufactured by Wako Pure Chemical Industries, Ltd.), MB752 /
1 (Waymouth's MB752 / 1, manufactured by Gibco), FBS (manufactured by Gibco), Paindex # 3 (manufactured by Matsutani Chemical Industry), ferric pyrophosphate (manufactured by Kokusan Chemical).

[0100]

Example 1 Production of freeze-dried powder of sweet tea water extract 1 kg of dried sweet tea powder (manufactured by Shihei Shoten) was distilled water 1
The mixture was extracted twice with 0 liter (agitated at room temperature for 1 hour), and the obtained extract was concentrated and freeze-dried to obtain 200 g of a freeze-dried powder of a sweet tea water extract.

Example 2 Production of freeze-dried powder of 60% ethanol aqueous solution of sweet tea 1 kg of dried sweet tea powder was added to 60% ethanol aqueous solution 1
The mixture was extracted twice with 0 liter (agitated at room temperature for 1 hour), and the obtained extract was concentrated and freeze-dried to obtain 6 g of sweet tea.
200 g of a freeze-dried powder of 0% ethanol aqueous solution extract was obtained.

Example 3 Production of freeze-dried powder of sweet tea acetone extract 1 kg of dried sweet tea powder was mixed with 10 l of acetone.
Extraction was repeated (at room temperature for 1 hour), and the obtained extract was concentrated and lyophilized to obtain 100 g of a lyophilized powder of acetone extract of sweet tea.

Example 4 Production of freeze-dried powder of sweet tea hot water extract 1 kg of dried sweet tea powder was extracted with 10 liters of boiling distilled water (left for 30 minutes), and the obtained extract was concentrated and freeze-dried. As a result, 130 g of a freeze-dried powder of a hot water extract of sweet tea was obtained.

Example 5 Preparation of freeze-dried powder of ethanol extract residue of water extract of sweet tea 1 kg of dried sweet tea powder was added to 20 liters of distilled water with 1 liter.
Extraction (40 ° C., stirring) was performed until the absorbance at 313 nm of the / 200 dilution became 0.15. The filtrate was removed by filtration to obtain an extraction residue. The residue is 60% ethanol 20
Extracted in liters until the absorbance at 313 nm of the 1/200 dilution was 0.22 (40 ° C., stirring). The extract was concentrated and then freeze-dried to obtain 70 g of a freeze-dried powder of ethanol extract residue extract of sweet tea in water (yield to dry leaf 7%).

Example 6 Production of Lyophilized Powder of Acetone Extract Residue from Aqueous Extract of Sweet Tea 1 kg of dried sweet tea powder was added to 20 liters of distilled water using
Extraction (40 ° C., stirring) was performed until the absorbance at 313 nm of the / 200 dilution became 0.15. The filtrate was removed by filtration to obtain an extraction residue. The residue was washed with 20 liters of acetone and the absorbance at 313 nm of a 1/200 dilution was 0.22.
(40 ° C, stirred) The extract was concentrated and then freeze-dried to obtain 60 g of a freeze-dried powder of acetone extract residue of sweet tea in water (yield to dry leaf 6%).

Example 7 Production of a feed containing 3% of the lyophilized powder of Example 1 A feed having the following composition was produced by mixing the components.

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) ) 37.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast) 3.5% by weight Cellulose (manufactured by Oriental Yeast) 5.0% by weight casein ( (Wako Pure Chemical Industries, Ltd.) 25.0% by weight Powder produced in Example 1 3.0% by weight

Example 8 Production of a feed containing 3% of the lyophilized powder of Example 2 A feed having the following composition was produced by mixing the components. .

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) ) 37.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast) 3.5% by weight Cellulose (manufactured by Oriental Yeast) 5.0% by weight casein ( (Wako Pure Chemical Industries, Ltd.) 25.0% by weight Powder produced in Example 2 3.0% by weight

Example 9 Preparation of feed containing 3% of freeze-dried powder of Example 4 A feed having the following composition was prepared by mixing the components. .

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) ) 37.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast) 3.5% by weight Cellulose (manufactured by Oriental Yeast) 5.0% by weight casein ( (Wako Pure Chemical Industries, Ltd.) 25.0% by weight Powder produced in Example 4 3.0% by weight

Example 10 Freeze-dried powder of Example 5 1%
Production of a feed containing the following: A feed having the following composition was produced by mixing each component.

Sucrose (manufactured by Kishida Chemical Co.) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) 39.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast Co., Ltd.) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast Co., Ltd.) 3.5% by weight Cellulose (manufactured by Oriental Yeast Co., Ltd.) 5.0% by weight casein ( 25.0% by weight Powder produced in Example 5 1.0% by weight

Comparative Example 1 A feed having the following composition was produced by mixing each component.

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) ) 40.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast) 3.5% by weight Cellulose (manufactured by Oriental Yeast) 5.0% by weight casein 25.0% by weight (manufactured by Wako Pure Chemical Industries)

Example 11 5 g of the lyophilized powder obtained in Example 1 was added to 50 parts of water.
The suspension was 0 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 25
Elute with 0 ml of 33% methanol and add 250 ml
The liquid eluted with 33% methanol was concentrated and dried to obtain a concentrated dried product (3). The column is 250m
and eluted with 66% methanol.
By drying, a concentrated dried product (4) was obtained. further,
The mixture was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrate (5).
The column was eluted with 250 ml of 100% methanol,
The eluted liquid was concentrated and dried to obtain a concentrated dried product (6). Further, elution was carried out with 250 ml of 100% methanol and then with 250 ml of 100% acetone, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (8). Then, the liquid eluted with 250 ml of 100% acetone was concentrated and dried to obtain a concentrated dried product (9).

Example 12 5 g of the freeze-dried powder obtained in Example 4 was mixed with 50 parts of water.
The suspension was 0 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 25
Elute with 0 ml of 33% methanol and add 250 ml
The liquid eluted with 33% methanol was concentrated and dried to obtain a concentrated dried product (3). The column is 250m
and eluted with 66% methanol.
By drying, a concentrated dried product (4) was obtained. further,
The mixture was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrate (5).
The column was eluted with 250 ml of 100% methanol,
The eluted liquid was concentrated and dried to obtain a concentrated dried product (6). Further, the liquid was eluted with 250 ml of 100% methanol, then eluted with 250 ml of 100% acetone, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (8). Then, the mixture was eluted with 250 ml of 100% acetone, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (9).

Example 13 5 g of the freeze-dried powder obtained in Example 2 was mixed with 50 parts of water.
The suspension was 0 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 25
Elute with 0 ml of 33% methanol and add 250 ml
The liquid eluted with 33% methanol was concentrated and dried to obtain a concentrated dried product (3). The column is 250m
and eluted with 66% methanol.
By drying, a concentrated dried product (4) was obtained. further,
The mixture was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrate (5).
The column was eluted with 250 ml of 100% methanol,
The eluted liquid was concentrated and dried to obtain a concentrated dried product (6). Further, the solution was eluted with 250 ml of 100% methanol, and the eluted solution was concentrated and dried to obtain a concentrated and dried product (7). Then 250ml 100%
After elution with acetone, elution was carried out with 250 ml of 100% acetone, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (9).

Example 14 5 g of the freeze-dried powder obtained in Example 5 was mixed with 50 parts of water.
The suspension was 0 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 25
Elute with 0 ml of 33% methanol and add 250 ml
The liquid eluted with 33% methanol was concentrated and dried to obtain a concentrated dried product (3). The column is 250m
and eluted with 66% methanol.
By drying, a concentrated dried product (4) was obtained. further,
The mixture was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrate (5).
The column was eluted with 250 ml of 100% methanol,
The eluted liquid was concentrated and dried to obtain a concentrated dried product (6). Further, the solution was eluted with 250 ml of 100% methanol, and the eluted solution was concentrated and dried to obtain a concentrated and dried product (7). Then 250ml 100%
After elution with acetone, elution was carried out with 250 ml of 100% acetone, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (9).

Example 15 5 g of a freeze-dried powder of the acetone extract obtained in Example 3 was suspended in 500 ml of water. The suspension was treated with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20,
(Void volume: 100 ml). Then, it was washed twice with 250 ml of 33% methanol. Then, it was washed twice with 250 ml of 66% methanol. Further, it was washed twice with 250 ml of 66% methanol. Then, after washing with 250 ml of 100% methanol, elution is performed with 250 ml of 100% acetone, and the eluted liquid is concentrated and dried to obtain a concentrated and dried product (8).
Got.

Example 16 Inhibition of D-galactosamine-induced rat liver injury by various sweet tea extracts Wistar purchased from SLC Japan
A white rat (male, 150 ± 20 g) was acclimated for 3 days or more under certain conditions (temperature: 24 ± 2 ° C., humidity: 60 ± 5%, light / dark cycle: 12 hours). The prepared feed (test group) and the feed prepared in Comparative Example 1 (control group) were each fed for a total of 15 days. 1
On the fourth day, D-galactosamine (dissolved in physiological saline at a concentration of 35 mg / ml and adjusted to pH 7.1) 3
50 mg / kg was administered intraperitoneally. Twenty-two hours after administration of D-galactosamine, the abdomen was opened under anesthesia with Nembutal and blood was collected.

Using the blood thus obtained, GPT activity in the blood was measured by the following method as an index of liver function. After coagulating the collected blood, it is centrifuged,
Serum was obtained. Using the obtained serum, GPT in the serum was measured using Transaminase CII-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The GPT activity was determined by calculating the relative value (%) of the value obtained with the feed (test group) of each Example, with the value of the control group taken as 100%. Values are shown as mean ± standard error, and statistical significance test was performed by T-test.

The results are shown in Table 1.

[0124]

[Table 1]

When the feeds of Examples 7 to 10 were administered, the GPT activity in serum, which is an indicator of liver dysfunction, was as low as 14.3% to 49.8% as compared to the case where the diets of Comparative Examples were administered. It can be seen that the hepatic disorder was suppressed. Also,
Since the GPT activity of the feed of Example 10 was 14.3%, it can be seen that the ethanol extract obtained by extracting the sweet tea water extracted in Example 5 has a strong liver injury suppressing action.

During the 15-day feeding period, no difference was observed in weight gain and no abnormal appearance or behavior was observed in any of the feeds.

Example 17 Inhibition of Acetaminophen-Induced Primary Culture Hepatocyte Damage by the Sweet Tea Acetone Extract Fraction A collagenase perfusion method [Seglen (PO), Methods in Cell Biol (Methods in Cell Biol)
ogy), 13 , 29 (1976)]. That is, a male SD weighing about 130 g
Rats were laparotomized under anesthesia and pre-perfused solution [Hanks' Balanced Salt Solution (manufactured by Gibco) 9.5 g), HEPES (Nacalai Tesque) 2.38 g, EGTA (Sigma) 0.19 g,
0.35 g of NaHCO ₃ (manufactured by Kishida Chemical Co., Ltd.) was dissolved in 1 L of water and adjusted to pH 7.2] by 30 mL from the portal vein.
400 ml was perfused at a flow rate of ml / min. Next, a collagenase solution [Hank's solution Nissui (manufactured by Nissui Pharmaceutical Co., Ltd.) 9.8 g, HEPES (manufactured by Nakarai Tesque) 2.38 g, NaHCO ₃ (manufactured by Kishida Chemical Co.) 0.
35 g, 0.56 g of CaCl ₂ (manufactured by Kishida Chemical), 0.02 g of trypsin inhibitor (manufactured by Sigma), and 0.5 g of collagenase (manufactured by Sigma) were dissolved in 1 L of water,
pH 7.5) was perfused. After the liver is digested, collect in a Petri dish and add 20 ml of S
-MEM medium (manufactured by Gibco) was added and cut into small pieces with a scalpel. Then pipette with a 10ml tip thick Komagome pipette,
After dispersing the hepatocytes, the hepatocytes were filtered with a gauze and a cell filter (manufactured by Ikemoto Rika Kogyo Co., Ltd.) to obtain a hepatocyte dispersion.
Hepatocytes obtained here are non-parenchymal cells other than hepatocytes,
For example, since it contains endothelial cells, Kupffer cells, Ito cells, stellate cells, etc., only hepatocytes were purified by centrifugation. Specifically, the obtained hepatocyte dispersion was subjected to 50 ×
G was centrifuged at a low speed under cooling for 1 minute to collect hepatocytes precipitated at the bottom. This operation was repeated three times to separate and collect hepatocytes with high purity.

The separated and recovered hepatocytes were suspended in the following basic medium to a concentration of 1.2 × 10 ⁶ cells / ml, and seeded on a 6-well (well) plate coated with Matrigel at 1.4 ml. Primary culture was performed under the following culture conditions. The culture medium was Waymouth's MB752 / Waymouth's MB752 /
1) In a medium (manufactured by Gibco), bovine serum (FBS, 10
%), Penicillin (50 U / ml), streptomycin (50 μg / ml), insulin (10 ⁻⁸ M), and dexamethasone (10 ⁻⁶ M) (hereinafter sometimes referred to as a basal medium). Was used.

The culture was carried out in a CO ₂ incubator at 37 ° C., 5% CO ₂ , 95% atmosphere. Four hours after the completion of the seeding, the basal medium was removed, and after washing with PBS, a new basal medium (0.7 ml) was dispensed and further cultured. The concentrated and dried product (8) obtained in Example 15 was treated with dimethyl sulfoxide (hereinafter abbreviated as DMSO) at 10 mg / ml.
And dissolved in the above-mentioned basic medium for 50 minutes.
Concentrate to a concentration of 200 μg / ml
Was prepared.

24 hours after cell seeding, 140 μl of test solution was added to each well (final concentration of the fraction: 20 μg / m 2).
1) One hour later, 560 μl of 50 mM acetaminophen (liver damage inducer) was added (final concentration: 20 mM).
(It will be a test area). Further, 24 hours after cell seeding, 140 μl of a basic medium containing 2% DMSO was added to each well, and 1 hour later, 560 μl of 50 mM acetaminophen (a liver injury inducer) was added (final concentration: 20 mM).
(This is the first control section).

Also, 24 hours after cell seeding, each well was
Was added with 2% DMSO-containing basic medium (140 μl), and one hour later, 560 μl of distilled water was added (a second control section). 48 hours after addition of acetaminophen or distilled water, MTT [3- (4,5-Dimethyl-2-thiazolyl)-
2,5-diphenyl-2H-tetrazolium bromide] assay, the number of cells in each section was counted by absorbance. Specifically, except for the basal medium in each well, MTT (10 m
g / ml) in 10% PBS containing Waymo
1.4 ml of ith's MB752 / 1 medium was dispensed.
After incubating for 1 hour in a CO ₂ incubator at 37 ° C., 4.2 ml of DMSO was added, and the mixture was vigorously stirred.
In Model 3550), the absorbance at 570 nm was measured. The evaluation was performed in duplicate. The hepatocellular injury inhibition rate was calculated by the following equation.

[0132]

(Equation 1)

The results are shown in Table 2.

[0134]

[Table 2]

As shown in Table 2, the concentrated dried product (8) obtained in Example 15 suppressed the hepatocyte damage caused by acetaminophen by 42.2%.

Example 18 Inhibition of D-Galactosamine-Induced Primary Culture Hepatocyte Damage by Fractions of Various Sweet Tea Extracts Concentrated and dried product (3) obtained in Examples 11 and 12,
(4), (5), (6), (8) and (9) and the concentrated and dried product (3) obtained in Examples 13 and 14,
(4), (5), (6) and (7) are each replaced with DMS
Dissolve with O to 10 mg / ml, and add 50
A dilution of a fraction diluted 1-fold (fraction concentration 200 μg / m
l) was prepared. Cells were seeded and primary cultured in the same manner as in Example 17. 2 hours after seeding, the medium was removed and PB
After washing with S, a new medium (0.7 ml) was dispensed,
140 μl of a fraction dilution was added (final concentration of fraction 20
μg / ml). Two hours later, 50 mM dissolved in the medium
560 μl of D-galactosamine (a liver injury inducer) was added (final concentration: 20 mM) (referred to as a test group).

After 2 hours from the cell seeding, 140 μl of a basic medium containing 2% DMSO was added to each well.
After a lapse of time, 560 μl of 50 mM D-galactosamine (a liver injury inducer) dissolved in the medium was added (final concentration 20 m).
M) (First control group). In addition, cell seeding 2
After an hour, each well was filled with a basic medium 14 containing 2% DMSO.
0 μl was added, and two hours later, 560 μl of the medium was added (a second control section).

48 hours after the addition of D-galactosamine or medium, the number of cells in each section was counted by absorbance by MTT assay. Specifically, except for the basal medium in each well, PBS containing MTT (10 mg / ml) was dissolved.
Was dispensed in 1.4 ml of Waymouth's MB752 / 1 medium containing 10%. After incubating for 1 hour in a CO ₂ incubator at 37 ° C, 4.2 ml of DM
After adding SO and stirring vigorously, the mixture was placed in a microplate reader (Bio-Rad, model 3550).
The absorbance at 570 nm was measured. The evaluation was performed in duplicate. The hepatocellular injury inhibition rate was calculated by the above equation (1).

Table 3 shows the results.

[0140]

[Table 3]

As shown in Table 3, fractions of various extracts of sweet tea showed hepatocellular damage due to D-galactosamine.
8 to 97.4% was suppressed.

Example 19 Inhibition of Primary Cultured Hepatocyte Damage Induced by Combination of D-Galactosamine and TNF-α by Fractions of Sweet Tea Extracts The concentrated dried product (9) obtained in Examples 12, 13 and 14 was Each was dissolved in DMSO to 10 mg / ml, and this was diluted 50-fold with a basal medium to obtain a fraction dilution liquid (fraction concentration: 500 μg / ml).

The cells were seeded and cultured in the same manner as in Example 17. Two hours after seeding, the medium was removed, washed with PBS, a new medium (0.7 ml) was dispensed, and 140 μl of a fraction dilution was added (final concentration of the fraction: 20 μg /
ml). Two hours later, 2.5 mM D-
560 μl of a mixed solution of galactosamine and 2.5 ng / ml TNF-α (hereinafter referred to as GT solution) was added (final concentration: 1 mM D-galactosamine, 1 ng / TNF-α).
ml) (referred to as test plot).

Two hours after cell seeding, 140 μl of a basic medium containing 2% DMSO was added to each well.
After a lapse of time, 560 μl of a GT solution (liver injury inducer) was added (final concentration: D-galactosamine 1 mM, TNF-α1n).
g / ml) (first control). Two hours after cell seeding, 140 μl of 2% DMSO-containing basic medium was added to each well, and two hours later, the medium was 560 μl in volume.
1 (the second control).

48 hours after the addition of the GT solution or the medium, the MTT
In the assay, the number of cells in each section was measured by absorbance. Specifically, except for the basal medium in each well,
1.4 m of Waymouth's MB752 / 1 medium containing 10% of PBS in which TT (10 mg / ml) is dissolved
1 was dispensed. After incubating for 1 hour in a CO ₂ incubator at 37 ° C., 4.2 ml of DMSO was added, and the mixture was vigorously stirred, and then 570 nm in a microplate reader (Bio-Rad, Model 3550).
Was measured for each absorbance. The evaluation was performed in duplicate. The hepatocellular injury inhibition rate was calculated according to the above equation 1.

[0146]

[Table 4]

As shown in Table 4, the fractions of the various extracts of sweet tea suppressed the hepatocellular damage caused by the combined use of D-galactosamine and TNF-α by 17.2 to 34.0%.

Example 20 A feed having the following composition was produced by mixing the components. CE-2 (manufactured by CLEA Japan) 99% by weight 1% by weight of the powder produced in Example 5

Example 21 A feed having the following composition was produced by mixing each component.

CE-2 (manufactured by CLEA Japan) 99% by weight 1% by weight of the powder produced in Example 3

Comparative Example 2 A feed having the following composition was produced by mixing each component. CE-2 (Clea Japan) 99% by weight Paindex # 3 (Matsuya Chemical) 1% by weight

Example 22 Alcohol / Lipopolysaccharide (LP)
S) Inhibition of induced rat liver injury SD white rats (female, 200
g) under constant conditions (temperature: 24 ± 2 ° C., humidity: 60)
After acclimation for 3 days or more at ± 5%, light-dark cycle: 12 hours), the feed of Example 20 (test group) or Comparative Example 2 (control group) was fed for a total of 15 days. Fifteen
During the one-day feeding period, no weight gain, appearance or behavioral abnormalities were observed between rats fed any diet. On the 14th day, alcohol 4 g / kg (40 v / v%
Ethanol solution) was orally administered using a sonde. Six hours later, 5 mg / kg of LPS (derived from E. coli , manufactured by Sigma) dissolved in physiological saline was administered by tail vein injection, and 24 hours later, laparotomy was performed under anesthesia with Nembutal. Blood was collected. The test was performed with 6 animals in each group.

Using the blood thus obtained, the activity of GPT and GOT in the blood was measured as an index of liver function.
It was measured by the following method. After coagulating the collected blood,
Centrifugation was performed to obtain serum. Using the obtained serum, GPT and GOT in the serum were measured using Fuji Dry Chem System 3500 (manufactured by Fujifilm Corporation). The GPT and GOT activities of the test group were determined by calculating relative values with the value of the control group taken as 100%. In addition, these experimental results were shown by the average value +/- standard error, and the statistical significance test was performed by T-test.

Table 5 shows the results.

[0155]

[Table 5]

GPT or GOT activity in serum, which is an indicator of liver dysfunction, was reduced to 10.8 and 7.7% in the test group to which the sweet tea extract was administered compared to the control group, indicating that the liver damage was suppressed. confirmed. In the significance test, p
<0.01, indicating a significant difference.

Example 23 D Using Sweet Tea Acetone Extract
-Inhibition of galactosamine-induced rat liver injury SD white rats (male, 150
± 20 g) under certain conditions (temperature: 24 ± 2 ° C, humidity: 6)
(0 ± 5%, light / dark cycle: 12 hours), and after acclimating for 3 days or more, the feed prepared in Example 21 (test group) and the feed prepared in Comparative Example 2 (control group) were each fed for 4 days. Was. After that, the animals were fasted for 18 hours and D-galactosamine (dissolved in physiological saline at a concentration of 40 mg / ml).
400 mg / kg was administered intraperitoneally. Twenty-two hours after administration of D-galactosamine, the abdomen was opened under anesthesia with Nembutal and blood was collected.

Using the blood thus obtained, GPT activity in the blood was measured by the following method as an indicator of liver function. After coagulating the collected blood, it is centrifuged,
Serum was obtained. Using the obtained serum, GPT in the serum was measured using Transaminase CII-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The GPT activity of the test group was determined by calculating a relative value with the value of the control group taken as 100. Values are shown as mean ± standard error, and statistical significance test was performed by T-test. The results are shown in Table 6.

[0159]

[Table 6]

When the diet of Example 21 was administered, the GPT activity in the serum, which is an indicator of liver dysfunction, was low at 56.4% as compared with the diet of Comparative Example 2, and liver injury was suppressed. . There were no weight gain, appearance or behavioral abnormalities during the feeding period.

Example 24 Production of freeze-dried powder of water extract of Sakinoshita 1 kg of dried Sakinoshita powder (manufactured by Shihei Shoten Co., Ltd.) was extracted twice with 10 liters of distilled water (at room temperature and stirred for 1 hour).
The obtained extract was concentrated and lyophilized to obtain 150 g of a lyophilized powder of a water extract of Saxifraga.

Example 25 Production of freeze-dried powder of saxifrage aqueous solution of 60% ethanol 1 kg of dried saxifrage powder was extracted twice with 10 liters of a 60% aqueous ethanol solution (room temperature, stirred for 1 hour).
The obtained extract was concentrated and freeze-dried to obtain 188 g of freeze-dried powder of a 60% aqueous ethanol extract of Saxifraga.

Example 26 Production of Lyophilized Powder of Saxifrage Acetone Extract 1 kg of dried saxifrage powder was extracted twice with 10 liters of acetone (room temperature, stirred for 1 hour), and the obtained extract was concentrated and freeze-dried. As a result, 164 g of a freeze-dried powder of a saxifrage acetone extract was obtained.

Example 27 Production of freeze-dried powder of Saxifrage hot water extract 1 kg of dried Saxifrage powder was distilled water 10
The resulting extract was concentrated and lyophilized to obtain 100 g of lyophilized powder of hot water extract of Saxifraga.

Example 28 5 g of the lyophilized powder obtained in Example 24 was
Suspended in 00 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 2
Elute with 50 ml of 33% methanol and add 250 m
The liquid eluted with 1 l of 33% methanol was concentrated and dried to obtain a concentrated dried product (3). 250 columns
The eluted solution was eluted with 66 ml of 66% methanol, and the eluted solution was concentrated and dried to obtain a concentrated and dried product (4). Further, the liquid was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (5).

Example 29 5 g of the lyophilized powder obtained in Example 25 was added to water 5
Suspended in 00 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 2
Elute with 50 ml of 33% methanol and add 250 m
The liquid eluted with 1 l of 33% methanol was concentrated and dried to obtain a concentrated dried product (3). 250 columns
The eluted solution was eluted with 66 ml of 66% methanol, and the eluted solution was concentrated and dried to obtain a concentrated and dried product (4). Further, the liquid was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (5). The column was eluted with 250 ml of 100% methanol, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (6).

Example 30 5 g of the freeze-dried powder obtained in Example 26 was added to water 5
Suspended in 00 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 2
The mixture was eluted with 50 ml of 33% methanol, and the eluted solution was concentrated and dried to obtain a concentrate (2). Elution with a further 250 ml of 33% methanol followed by 2
The mixture was eluted with 50 ml of 66% methanol, and the eluted solution was concentrated and dried to obtain a concentrate (4).

Example 31 5 g of the lyophilized powder obtained in Example 27 was added to water 5
Suspended in 00 ml. The suspension was passed through a column filled with a hydrophobic adsorption resin (manufactured by Mitsubishi Chemical Corporation, Diaion HP-20, void volume 100 ml). Then 2
Elution was carried out twice with 50 ml of 33% methanol. The column was eluted with 250 ml of 66% methanol, and the eluted liquid was concentrated and dried to obtain a concentrated dried product (4). Further elute with 250 ml of 66% methanol,
The eluted liquid was concentrated and dried to obtain a concentrated dried product (5).

Example 32 A feed having the following composition was produced by mixing the components.

CE-2 (manufactured by CLEA Japan, Ltd.) 99.0% by weight Powder prepared in Example 24 1.0% by weight

Example 33 A feed having the following composition was produced by mixing the components. CE-2 (manufactured by CLEA Japan) 99.0% by weight Powder produced in Example 25 1.0% by weight

Example 34 Inhibition of D-galactosamine-induced liver injury in rats by Saxifrage extract [Saxifrage water extract (Example 24) and 60% ethanol extract (Example 25)] SD white rats (male) purchased from SLC of Japan , 150
± 20 g) under certain conditions (temperature: 24 ± 2 ° C, humidity: 6)
(0 ± 5%, light-dark cycle: 12 hours), and after acclimating for 3 days or more, feeds prepared in Examples 32 and 33 (test group) and feeds prepared in Comparative Example 2 (control group) for 3 days. They were fed. Thereafter, the animals were fasted for 18 hours, and 400 mg / kg of D-galactosamine (dissolved in physiological saline at a concentration of 40 mg / ml) was intraperitoneally administered. Further, after the above-mentioned feed was ingested for one day, the abdomen was opened under anesthesia with Nembutal and blood was collected.

Using the blood thus obtained, GPT activity in the blood was measured by the following method as an index of liver function. After coagulating the collected blood, it is centrifuged,
Serum was obtained. Using the obtained serum, GPT in the serum was measured using Transaminase CII-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The GPT activity was determined by calculating the relative value (%) of the value of the test group, taking the value of the control group as 100%. The results of these experiments are shown as mean ± standard error, and the statistical significance test was performed by T-test. The results are shown in Table 7.

[0174]

[Table 7]

When the feeds of Examples 32 and 33 were administered, the GPT activity in the serum, which is an indicator of liver dysfunction, was 3 compared to the case where the feed of Comparative Example 2 was administered.
It was confirmed that liver damage was suppressed as low as 5.7 and 29.9%. As a result of the significant difference test, a significant difference was recognized. No weight gain, no apparent, and no behavioral abnormalities were observed during the feeding period.

Example 35 Inhibition of Acetaminophen-Induced Primary Culture Hepatocyte Damage by Sacrificial Acetone Extract Fraction Using the concentrated and dried products (2) and (4) of Example 30,
The test was performed in the same manner as in Example 17. The results are shown in Table 8.

[0177]

[Table 8]

As shown in Table 8, the fractions of the extract of Sakinoshita in acetone inhibited hepatocellular damage caused by acetaminophen by 46.7% and 46.4%, respectively.

Example 36 Inhibition of D-galactosamine-induced primary culture hepatocyte damage by various extract fractions of Saxifraga was carried out in the same manner as in Example 18 using the concentrated and dried products of Examples 28, 29, 30, and 31. Tested.

Table 9 shows the results.

[0181]

[Table 9]

As shown in Table 9, the fractions of various extracts of Saxifraga inhibited D-galactosamine-induced hepatocellular damage by 36.1 to 111.1%.

Example 37 Inhibition of Primary Hepatocyte Damage Induced by Combined Use of D-Galactosamine and TNF-α by Various Extract Fractions of Saxifraga Example 19 Using the Concentrated Drying Materials Prepared in Examples 28, 29 and 31 The same test was performed. The results are shown in Table 10.

[0184]

[Table 10]

As shown in Table 10, the fractions of various extracts of Saxifraga inhibited the hepatocellular damage by D-galactosamine and TNF-α by 17.0 to 90.6%.

Example 38 Production of a Combination of an Ethanol Extract Residue from Sweet Tea Water Extract A liver function protector or liver function improver having the following composition was prepared.
It was produced by mixing the components.

Ethanol extract of sweet tea water extract residue produced in Example 5 49 g Paindex # 3 49 g Ferric pyrophosphate (iron source) 0.1 g Foscal EFC (calcium source; manufactured by Nikko Fine Products) 1 g Vitamin mix (Merck 1g

Example 39 20 g of the liver function protecting or improving agent prepared in Example 38
Was dispersed in 180 ml of water to produce a beverage for protecting or improving liver function.

Example 40 Production of Cookies Containing Ethanol Extract Residue from Sweet Tea Water Extraction Cookies (30 pieces) were produced by a conventional method according to the following composition.

Soft flour 100 g Starch 74 g Water 14 g Ethanol extract of sweet tea water extract residue prepared in Example 5 30 g Baking powder 2 teaspoon salt 2 teaspoon 1 egg butter 80 g Milk 2 tablespoon Example 41 1% of freeze-dried powder of Example 24 Production of Feedstuffs Containing Feedstuffs having the following composition were produced by mixing each component.

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Chemical Industry Co., Ltd.) 0.4% by weight Corn starch (manufactured by Nisseki Chemical Co., Ltd.) 39.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast Co., Ltd.) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast Co., Ltd.) 3.5% by weight Cellulose (manufactured by Oriental Yeast Co., Ltd.) 5.0% by weight casein ( 25.0% by weight Powder produced in Example 24 1.0% by weight

Example 42 Freeze-dried powder 1 of Example 25
% Of feed containing the following ingredients: A feed having the following composition was prepared by mixing the components.

Sucrose (manufactured by Kishida Chemical) 20.0% by weight Corn oil (manufactured by Nacalai Tesque) 5.0% by weight Choline bitartrate (manufactured by Tokyo Kasei Kogyo) 0.4% by weight Corn starch (manufactured by Nisseki Chemical) 39.1% by weight AIN-76 vitamin (manufactured by Oriental Yeast Co., Ltd.) 1.0% by weight AIN-76 mineral (manufactured by Oriental Yeast Co., Ltd.) 3.5% by weight Cellulose (manufactured by Oriental Yeast Co., Ltd.) 5.0% by weight casein ( 25.0% by weight Powder produced in Example 25 1.0% by weight

[0194]

Industrial Applicability According to the present invention, a liver function protecting or improving agent, a food or drink or feed having a liver function protecting or improving effect, a food or drink additive or a feed additive having a liver function protecting or improving effect. And a method for screening a liver function protecting agent or an improving agent.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 1/16 A61P 1/16 25/32 25/32 G01N 33/15 G01N 33/15 Z 33/50 33 / 50 Z (72) Inventor Yasushi Sakai 4041 Ami-cho, Ami-cho, Inashiki-gun, Ibaraki Prefecture Kyowa Hakko Kogyo Co., Ltd. F term in Tsukuba Research Laboratories (reference) 2B005 DA01 GA01 LB07 2B150 AA01 AA02 AA03 AA05 AA06 AA08 AA20 AB20 DD31 DD57 2G045 AA24 AA28 AA29 BA13 BB03 BB20 CA25 CB17 CB20 DA80 FB01 4B018 CA61 Z06MA06 Z06

Claims (51)

[Claims] 1. A liver function-protecting or improving agent comprising a plant of the family Saxifragaceae or an extract of the plant as an active ingredient. 2. The agent for protecting or improving liver function according to claim 1, wherein the plant of the family Saxifragaceae belongs to the genus Saxifraga. 3. The agent for protecting or improving liver function according to claim 2, wherein the plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb. 4. The liver function-protecting or improving agent according to claim 1, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 5. A plant belonging to the genus Hydrangea, wherein the plant belonging to the genus Hydrangea macrophylla Seringe var. Thunbergii Mak
The hepatic function-protecting or improving agent according to claim 4, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 6. The liver function-protecting or improving agent according to any one of claims 1 to 5, wherein the Saxifragaceae plant extract is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant. 7. The liver function-protecting or ameliorating agent according to claim 1, which is administered orally. 8. The liver function protecting or improving agent according to claim 1, wherein the liver function is a function affected by alcohol. 9. A food or drink to which a Saxifragaceae plant or an extract of the plant is added. 10. The food or drink according to claim 9, wherein the food or drink is used for protecting or improving liver function. 11. The food or drink according to claim 10, wherein the liver function is affected by alcohol. 12. The food or drink product according to claim 9, wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga. 13. The plant belonging to the genus Saxifraga is the saxifraga ( Saxifraga stolonifera Meerb.).
Food and drink according to 2. 14. The food or drink product according to claim 9, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 15. A plant belonging to the genus Hydrangea, wherein the plant belonging to the genus Hydrangea macrophylla Seringe var. Thunbergii Mak
The food or drink according to claim 14, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 16. The food or drink product according to any one of claims 9 to 15, wherein the Saxifragaceae plant extract is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant. 17. A feed to which a plant of the family Saxifragaceae or an extract of the plant is added. 18. The feed according to claim 17, wherein the feed is used for protecting or improving liver function. 19. The feed according to claim 18, wherein the liver function is a function affected by alcohol. 20. The feed according to any one of claims 17 to 19, wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga. 21. The plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.
0 feed. 22. The feed according to any one of claims 17 to 19, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 23. The plant belonging to the genus Hydrangea is an amateur ( Hydrangea macrophylla Seringe var. Thunbergii Mak)
The feed according to claim 22, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 24. The feed according to any one of claims 17 to 23, wherein the extract of the plant Saxifragaceae is an alcohol extract obtained from an aqueous medium extract of the plant Saxifragaceae. 25. An additive for foods and drinks having a hepatic function-protecting or ameliorating action, comprising a plant of the family Saxifragaceae or an extract of the plant. 26. The additive according to claim 25, wherein the liver function is a function affected by alcohol. 27. The additive for food and drink according to claim 25 or 26, wherein the Saxifragaceae plant is a plant belonging to the genus Saxifraga. 28. The plant belonging to the genus Saxifraga is Saxifraga stolonifera Meerb.
8. The additive for food and drink according to 7. 29. The additive for food or drink according to claim 25 or 26, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 30. A plant belonging to the genus Hydrangea, wherein the plant belonging to the genus Hydrangea macrophylla Seringe var. Thunbergii Mak
30. The additive for food and drink according to claim 29, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 31. The additive for food and drink according to any one of claims 25 to 30, wherein the extract of Saxifragaceae plant is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant. 32. A feed additive having a hepatic function-protecting or improving action, comprising a Saxifragaceae plant or an extract of the plant. 33. The feed additive according to claim 32, wherein the liver function is a function affected by alcohol. 34. The feed additive according to claim 32 or 33, wherein the Saxifragaceae plant is a plant belonging to the genus Saxifraga. 35. The plant belonging to the genus Saxifraga is the saxifraga ( Saxifraga stolonifera Meerb.).
4. The feed additive according to item 4. 36. The feed additive according to claim 32 or 33, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 37. A plant belonging to the genus Hydrangea, wherein the plant is a sea bream ( Hydrangea macrophylla Seringe var. Thunbergii Mak).
The feed additive according to claim 36, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 38. The feed additive according to any one of claims 32 to 37, wherein the Saxifragaceae plant extract is an alcohol extract of an aqueous medium extract residue of the Saxifragaceae plant. 39. GPT or GO in the blood of an animal which is increased by administration of a lipopolysaccharide after administration of alcohol by administering the substance to be screened to the animal.
A method for screening for an agent for protecting or improving liver function, which comprises evaluating a substance that decreases the T value. 40. The method according to claim 39, wherein the animal is a mammal. 41. The method according to claim 39 or 40, wherein the lipopolysaccharide is derived from a microorganism belonging to the group of intestinal bacteria. 42. The method according to claim 39, wherein the liver function is a function affected by alcohol. 43. A method for protecting the liver function of an animal, comprising feeding the animal with the agent for protecting or improving liver function according to any one of claims 1 to 8 or the feed according to any one of claims 17 to 24. Or how to improve liver function. 44. The method according to claim 43, wherein the animal is livestock, poultry, or farmed fish. 45. A food or drink or feed for protecting or improving liver function, which comprises a plant of the family Saxifragaceae or an extract of the plant as an active ingredient. 46. The food or drink or feed for protecting or improving liver function according to claim 45, wherein the saxifragaceae plant is a plant belonging to the genus Saxifraga. 47. The plant belonging to the genus Saxifraga is the saxifraga ( Saxifraga stolonifera Meerb.).
The food or drink or feed for protecting or improving liver function according to the above. 48. The food or drink or feed for protecting or improving hepatic function according to claim 45, wherein the Saxifragaceae plant is a plant belonging to the genus Hydrangea. 49. The plant belonging to the genus Hydrangea is an amateur ( Hydrangea macrophylla Seringe var. Thunbergii Mak)
49. The food or drink or feed for protecting or improving liver function according to claim 48, which is ino) or sweet tea (Hydrangeae Dulcis Folium). 50. The food or drink or feed for protecting or improving hepatic function according to any one of claims 45 to 49, wherein the extract of the plant Saxifragaceae is an alcohol extract obtained from an aqueous medium extract of the plant Saxifragaceae. . 51. The food, drink or feed for protecting or improving liver function according to any of claims 45 to 50, wherein the liver function is a function affected by alcohol.