JPH01242530A - Fatty liver suppressing substance - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION "Field of Industrial Application" The present invention relates to a fatty liver inhibitor that suppresses various liver function abnormalities associated with fatty liver.

"Prior Art" Fatty liver refers to a state in which fat is abnormally accumulated within liver cells, and the accumulated fat is mainly neutral fat. Although an extremely large number of factors are known to cause fatty liver, common factors are involved in its developmental mechanism. In other words, the liver plays an important role in fat metabolism and transfer.
Under normal conditions, fat balance in the liver is maintained at a certain equilibrium. Neutral fat in the liver consists of fatty acids in chylomicrons derived from meals and f1 mobilized from peripheral adipose tissue.
It is derived from 1 fatty acids and fatty acids newly synthesized in the liver. Some of the fatty acids newly synthesized or transferred to the liver are oxidized and degraded in the liver, but some are esterified in the liver microseams to become neutral fats, which combine with proteins and become liposomal fats. It is secreted into the bloodstream in the form of protein.

The turnover of neutral fats is rapid compared to other fats, and intravenously administered labeled free fatty acids are already found in lipoproteins in the bloodstream within 20 minutes. In this way, hepatic neutral fats are maintained in an equilibrium state through relatively rapid metabolism and turnover, but this equilibrium is disturbed, or in other words, an inconsistency occurs between the amount of neutral fats produced and utilized in the liver. Fatty liver occurs when these disorders are added.

Fatty liver is often caused by excessive intake of alcohol, obesity, and diabetes.
Patients complain of weight loss, fatigue, etc. Fatty liver also appears as an early symptom of liver damage such as fatty liver cirrhosis. In addition, in animal experiments such as rats, we artificially created fatty liver by administering ethionine, carbon tetrachloride, orotic acid, etc., and investigated the mechanism of fatty liver development.
Attempts are also being made to investigate methods for suppressing fatty liver.

However, the mainstream treatment for fatty liver is so-called diet therapy, which involves refraining from drinking alcohol and being careful about what you eat, but a method to proactively treat this has yet to be found. This is the current situation.

In addition, the present inventors studied the physiologically active effects of dietary fibers extracted from the outer skins of grains and beans, and found that these have the effect of activating liver function. A patent application has already been filed (Japanese Unexamined Patent Publication No. 62-201
No. 620 and Japanese Patent Application No. 61-282913). These have the effect of suppressing the increase in GOT and GPT in the blood in an experiment in which D-galactosamine (a substance 't1 that artificially induces liver damage) was administered to rats.

However, there are actually many types of liver diseases, each with a different expression mechanism, so it is not well understood which liver diseases these dietary fibers or the hemicellulose obtained from them are particularly effective against. There wasn't. Furthermore, it was unclear whether all of the various dietary fibers obtained from the husks of grains and beans exhibit similar effects, or whether some of them exhibit particularly excellent effects.

[Problems to be Solved by the Invention] The present invention has been made in view of the above-mentioned problems of the prior art. The objective is to provide substances with action.

"Means for Solving the Problems" The fatty liver inhibitor according to the present invention has hemicellulose obtained from corn bran or wheat bran as a main component.

The fatty liver inhibitor according to the present invention is preferably obtained by removing starch, protein, etc. from corn bran or wheat bran, and extracting the remainder with an alkali.

"Effect" In an experiment in which orotic acid was administered to rats to artificially cause fatty liver, the present inventors investigated the fatty liver suppressing effect of the dietary fibers of each type of oar and the hemicellulose obtained from them. In particular, it was discovered that hemicellulose obtained from corn bran or wheat bran has an excellent effect of inhibiting fatty liver, which led to the completion of the present invention.

In this case, 1. Fatty liver in humans is not caused by orotic acid, but as mentioned above, common factors are involved in the mechanism of fatty liver development. It is also expected to be effective in suppressing fatty liver disease in humans.

And hemicellulose obtained from corn bran or wheat bran is a completely safe substance for the human body.
It can be safely ingested as a medicine, food additive, or food or beverage.

"Preferred Embodiment of the Invention" The fatty liver inhibitor of the present invention may have hemicellulose obtained from corn bran or wheat bran as its main component, and any method may be used to extract hemicellulose. . For example, extraction with alkali or acid, pressurization with an extruder or autoclave, hot water extraction, extraction with enzymes such as cellulase, etc.
Alternatively, any suitable combination of these methods may be employed.

However, in one preferred embodiment of the invention, corn bran or wheat bran is starchier.

Proteins, etc. are removed, and the remainder is extracted with alkali (1), which is used.

A preferred method for producing the fatty liver inhibitor of the present invention is as follows.

Corn bran or wheat bran can be directly extracted with alkaline to extract components rich in hemicellulose, but in order to obtain hemicellulose with higher purity, it is necessary to extract starch, protein, and other necessary components from corn or wheat bran. After removing lipids, inorganic substances, etc., it is preferable to carry out alkali extraction.

As a method for removing starch, protein, and, if necessary, lipids, inorganic substances, etc. from corn bran or wheat bran, any of enzyme treatment, chemical treatment, and physical treatment may be employed, or any of these may be used as appropriate. The six enzyme treatments that may be combined include, for example, starch-degrading enzymes such as α-amylase and glucoamylase, proteolytic enzymes such as protease, lipid-degrading enzymes such as lipase, and fibrillolytic enzymes such as cellulase at pH 3 to 9 and temperature. This is carried out by adding an additive under conditions of 30 to 100°C. In addition, chemical treatment can be carried out, for example, by adding an aqueous solution of mineral acid or organic acid to corn bran or wheat bran and heating it under conditions of pH 2 to 5, or by adding a food-grade surfactant to pH 3 to 8. This is done by heat treatment under the following conditions. Further, the physical treatment is carried out, for example, by pulverizing corn bran or wheat bran with a pulverizer such as a homogenizer or a hammer mill, and then sieving the resulting product.

After the corn bran or wheat bran has been treated in this way, alkaline extraction is carried out. Alkaline extraction is carried out by adding and mixing an alkaline aqueous solution such as an aqueous sodium hydroxide solution to elute the non-cellulosic polysaccharide fraction. The fatty liver inhibitor of the present invention is
This alkaline extract can be neutralized to obtain an unpurified product, but this neutralized product can also be further purified by the following procedure (i.e., the proteins precipitated by neutralization are centrifuged). It is separated and removed by means such as separation, and if necessary, the supernatant liquid is subjected to dialysis, ion exchange resin treatment,
Ion exchange membrane treatment, ultrafiltration membrane treatment, alcohol purification,
The fatty liver inhibitor of the present invention containing a hemicellulose component of any purity can be obtained by processing alone or in appropriate combinations such as filter media treatment.

The fatty liver inhibitor of the present invention obtained as described above is
The main component is hemicellulose, with some lignin,
It consists of things that contain cellulose, ash, etc.

The fatty liver inhibitor of the present invention can be commercialized as an extract, an aqueous solution, or a dry powder. Even when it is made into a dry powder, it is very soluble in water (it easily becomes a homogeneous solution, so it is easy to prepare. Therefore, it can be used as it is as health drinks and medicines, and it can also be added in small amounts to foods and drinks. By doing so, it is possible to impart physiological activity without impairing the flavor or texture of the food or drink.

"Example of the invention" (Experimental method) (1) Feed The feed composition is shown in Table 1. The standard feed consists of sucrose as a carbohydrate source, milk casein as a protein source, corn oil as a lipid, and 1lar as a mineral mixture and a vitamin mixture.
A formulation of per was used. 1% orotic acid was added to all samples other than the standard group. In addition, the indicated dietary fibers were added to the groups other than the standard group and the control group. In addition,
In the group that added orotic acid and dietary fiber, some of the sucrose was reduced instead. The dietary fibers used here were prepared as follows.

■Wheat bran (hard type, manufactured by Nisshin Seifun) Wheat bran (
A hard type (manufactured by Nisshin Seifun ■) was stirred and washed in a mixer using distilled water to remove residual pesticides and increase the NDF value to 847%, and was sieved into 30 meshes for use. 1 equivalent to 5% of NDF was added to the feed.

■Corn bran After processing an aqueous dispersion of corn bran (solid content 10%) obtained by wet milling corn with a homogenizer for 1 minute, it was washed with water using a 48 mesh 1 sieve, separated by a sieve, and the The residue was collected, dried and powdered and used. The feed was supplemented with 5% Gogo as NDF.

■Corn hemicellulose 100g of corn bran prepared in the above ■512
Transfer to a large Erlenmeyer flask. Glucoamylase (manufactured by Nagashio Sangyo ■, l x 10' GUN/g15 g was dissolved in distilled water 4a, filtered through a filter paper, and 0.2M acetate buffer (PH4,8) II2 was added to the filtrate to prepare glucoamylase. Solution 52 and several drops of toluene were added to the above cone fiber and kept at 40°C for 24 hours. The mixture was filtered with a glass filter (151G3), the residue was washed with water, and transferred to a 2β Erlenmeyer flask, and 0.5N Add sodium hydroxide solution 1e, fill the container with nitrogen gas, seal it with a rubber stopper, and let it rise at room temperature for 18 hours (130 strokes/
minute) by making an alkali-soluble non-cellulosic poly-RF? We extracted the category i category. This was centrifuged (3000 rpm, 10 minutes) and neutralized with liquid glacial acetic acid.
Proteins were precipitated by adding trichloroacetic acid to a final moisture content of 7%.

The precipitate was removed by centrifugation (5000 rpm, 10 minutes), water was added to the resulting separated liquid (about 700 mI2.) to make about 1.5 g, and the mixture was placed in running water for 3 days using a cellophane tube. Dialyzed. After confirming that the dialyzed content has become neutral, add approximately 4 times as much ethyl alcohol (final concentration 8.
0% or more) was added, and the mixture was heated overnight to sufficiently form a precipitate. This precipitate was centrifuged (4000 rpm, 10
2 minutes), dissolved in distilled water 1e, and lyophilized to obtain 13 g of a light-colored powder. This powder has a moisture content of 12.8%
, ash], 9%, total nitrogen 0.8%, hemicellulose 83
0%, cellulose 01%, and lignin 14%. 2% of the corn hemicellulose thus obtained was added to the feed.

■Pectin High methoxyl pectin (degree of esterification 65%, Unip
ectine), and 2% of this was added to the feed.

■Wheat bran hemicellulose Wheat bran prepared using ■ above! 00 g, 1β water added with 2% calcium hydroxide was added, and hemicellulose was extracted by heating at 80 °C for 5 hours. 6 Centrifugation (500 g)
After removing the residue (0 rpm, 10 minutes), the extract was neutralized with sulfuric acid, filtered and decolorized, desalted for 1 minute using an ultrafiltration membrane with a molecular weight of 100,000, concentrated, and then lyophilized to obtain hemicellulose. 6.4 g of powder was obtained. This powder has a moisture content of 7.3% and an ash content of 4.
．． 1%, total nitrogen 0.2%, hemicellulose 84.2%,
It consists of 3% cellulose and 57% lignin. 2% of the wheat bran hemicellulose obtained in this way is used as feed.
Added.

(Hereinafter, blank spaces) (2) Experimental animals and breeding method The experimental animals used were SD male and female rats aged 4i4 (body weight 69 to 77 g). After preliminarily feeding for 6 days with standard diet, rats were fed 7 days so that the average body weight was equal for each P1.
The animals were divided into 6 groups, and each test animal was given 1 dose and kept for 11 days. During this period, feed and water were available ad libitum at zero body weight, and feed intake was measured at a fixed time every day. Main breeding 12
On the following day, the animals were anesthetized with intracostal injection of Nembutal at 2:00 p.m. without fasting, blood was collected by cardiac puncture, and dissection was performed.

(3) Analysis of serum and liver ■Analysis of biochemical values of serum The blood collected from the heart was immediately heated at 2000 rpm for 20
Serum was separated by centrifugation for one minute and analyzed for the following six items.

@Go”11 Glutamic acid/Oxaloacetate/Transaminase) Test reagent “Iatrozyme TA-LQJ” manufactured by Yatron■
(Product name) was used.

■GPT (glutamic acid/pyruvic acid/transaminase) Test reagent “Iatrozyme TA-LQJ” manufactured by Yatron ■
(Product name) was used.

0TC (total cholesterol) Kyowa Medex (Test reagent “Determiner TC5J” manufactured by Kiki)
(Product name) was used.

■TG (triglyceride/neutral fat) Test reagent "Determiner-TG-3555J (trade name)" manufactured by Kyowa Medex ■ was used.

@PL (phospholipid) Kyowa Medex (… test reagent “Decuminar PLJ”)
(Product name) was used.

■OCT (ornithine carbamyl transferase) ■Analysis of liver lipid components Immediately after removal, the liver was weighed and freeze-dried.

After measuring the weight of the dried liver, crush it and homogenize it.
This was used as an analysis sample. 05g of the sample was weighed out and
Lipids were extracted according to the method of et al. That is, 5
Place sample 11 in a 0+nj 2 volume centrifuge tube with a stopper, add 15 m, c of a chloroform/methanol mixture (21, V/V), and stir with a thermomixer for 3 times every 30 minutes. After leaving it for 24 hours, centrifugation was performed at 3000 rpm for 10 minutes, and the supernatant liquid was filtered using filter paper (5
B), and the filtrate was brought to a constant volume of 50 mε with a chloroform/methanol mixture and subjected to the following four analysis items.

■Weigh out 20mj2 of the total lipid sample solution, put it into a flask with a known constant temperature, evaporate it to dryness on a hot water bath (90℃), and then place it in a dryer (90℃).
5℃), dried until it reached Ishikawa, and weighed to determine the lipid content.

(Total cholesterol (TC)) Take 1 ml of the sample solution, wash it with physiological saline, and use the test reagent [Determiner TC55] manufactured by Kyowa Medex ■.
The analysis was performed using 5J (trade name).

■Triglyceride (TG) Weigh out 1 mρ of the test solution, wash it with physiological saline, and then
Test test TX manufactured by Wako Junsou■: rTriglyceri
The analysis was performed using deTest™akoJf (trade name).

■Phospholipid (PL) Weigh out 1 mε of the test solution, wash it with physiological saline, and then
Test reagent “Determiner PL55” manufactured by Kyowa Medex [m]
5J (trade name) was used for analysis.

(4) Statistical processing Each measured value was subjected to multiple comparisons using an analysis of variance using a negative device, and significant differences were tested at P<0.05 (95% confidence probability).

(Experimental result) (+) Growth results are shown in Table 2.

No significant differences were observed between groups in body weight gain, feed efficiency, and eviscerated layer body weight.

For the liver type, each group with orotic acid added was treated with F without orotic acid.
g showed a significantly higher value than the subgroup (ST group) (P
<0.05), and among the groups to which orotic acid was added, the corn hemicellulose group and the wheat bran hemicellulose group showed significantly lower values than the control group.

In addition, the liver carcass weight ratio in each group with orotic acid addition was higher than that in the standard group, and a significant difference was observed (P<0
．． 05).

(Hereinafter, blank space) +21 ('X) Serum biochemical values of each test group are shown in Table 3.

Among the orotic acid-added groups, GOT contains wheat bran group,
The corn bran group, corn hemicellulose group, and wheat bran hemicellulose group showed significantly lower values than the control group (CP<0.05), and especially the corn bran group, corn hemicellulose group, and wheat bran hemicellulose group showed significantly lower values than the control group. No significant difference was observed between the two (Pr0.05). The pectin group was not different from the 2 control groups and significantly higher f than the corn hemicellulose group.
1α (P<o, 05).

No significant difference in GPT was observed between each group, regardless of the addition of orotic acid 1 dietary fiber.

Among the orotic acid-added groups, the TC was 61 for all groups other than the corn hemicellulose group and the wheat bran hemicellulose group. , showed a significantly lower value than the group (P<
o, 05), whereas the corn hemicellulose group and the wheat bran hemicellulose group are paired! It showed a significantly higher value than the IQ group, wheat bran group, and pectin group (P
<0.05+.

Among the orotic acid-added groups, the groups other than the corn hemicellulose group and the wheat bran hemicellulose group showed significantly lower TG values than the standard group.Pro, 051
, the corn hemicellulose group and the wheat bran hemicellulose group showed values so high that no significant difference was observed from the standard group (P < 0.05), and the PL of each group with orotic acid addition was significantly lower than the standard group. However, CP<o, 0
5) Among the orotic acid-added groups, the corn hemicellulose group and the wheat bran hemicellulose group showed significantly higher values than the other groups (pro, 05).

There was no significant difference in OCT between each dietary fiber addition group and the In group (IP<0.05), and the wheat bran group, corn bran group, corn hemicellulose group, and wheat bran hemicellulose group were all compared to the control group. significantly lower (1α) (p<0.051゜ (see margin below) It has been reported that when feed containing 1% orotic acid is administered to male rats, TC and PL decrease to about 1/2, TG to about 1/3, and GOT shows a slight increase.6 In the control group administered with 1% orotic acid-containing feed in the experiment, the serum chemical values showed almost the same tendency as above.On the other hand, the results of administering each dietary fiber in combination with orotic acid are as follows. , corn hemicellulose group, and wheat bran hemicellulose group suppressed the increase in GOT and TC.

It was observed that the decrease in lipid components of PL and TG was significantly suppressed. In contrast, none of the above effects were observed in the pectin group, which is a water-soluble dietary fiber similar to corn hemicellulose. Furthermore, in the wheat bran group and corn bran group, the increase in GOT was significantly suppressed, but TC and PL values were slightly higher than in the control group, indicating a tendency to suppress the decrease in serum lipids caused by orotic acid. Indicated.

(3) Next, Table 4 shows the analysis results of liver lipid components in each test group.
Shown below.

Regarding total lipids in the liver, each group to which orotic acid was added showed a significantly higher value than the standard group (P<0.05>, indicating fatty liver formation). The wheat bran group and pectin group showed significantly higher values than the control group (P < 0.05), and no difference was observed between the l-corn bran group, corn hemicellulose group, and wheat bran hemicellulose group. .

When looking at the feed intake of each test group, each group with dietary fiber addition had a higher feed intake and, therefore, a higher intake of orotic acid, which resulted in a higher liver lipid content. it is conceivable that. There was no significant difference in hepatic lipid content between the corn bran group and the corn hemicellulose group compared to the control group, and in particular, the corn hemicellulose group and the wheat bran hemicellulose group had lower levels of orotic acid, even though they had a higher intake than the control group. showed that.

The biochemical characteristics of fatty liver caused by orotic acid administration are TG fatty liver that starts from the periphery of the liver stent, and it has been reported that abnormal accumulation of TG is observed in rats administered orotic acid-containing feed. In this experiment as well, the liver TG of the control group to which orotic acid was administered showed high values. As a result of administering each dietary fiber in combination with orotic acid, the corn hemicellulose group showed the lowest value in hepatic TG accumulation. this is.

Corn hemicellulose has T compared to other dietary fibers.
This is considered to be evidence that G accumulation is suppressed to some extent.

In addition, wheat bran group and wheat bran hemicellulose group are also T
G showed a relatively low value.

On the other hand, pectin, which is a water-soluble dietary fiber similar to corn hemicellulose and wheat bran hemicellulose, accumulated significantly more liver TG than the control group (P<0
．． 05), [Even from the abnormally low level of TG in flI serum, no effect of orotic acid in suppressing the formation of fatty liver was observed.

(Hereinafter, margins) From the above results, corn hemicellulose and wheat bran hemicellulose are effective against fatty liver formation in rats due to orotic acid administration, as well as GOT, TC, and PI-
When considered from the TG value, the total lipid content of the liver, and the TG value, it was found to have an excellent anti-fatty liver effect.

"Effects of the Invention" As explained above, the fatty liver inhibitor of the present invention has an excellent fatty liver inhibitory effect in an experiment in which orotic acid was administered to rats. therefore.

It is also expected to have an excellent effect on suppressing fatty liver in humans.

Furthermore, since the fatty liver inhibitor of the present invention is made of hemicellulose obtained from corn bran or wheat bran, it is a completely safe substance for the human body and can be safely ingested as a medicine, food additive, or food or beverage. .

Patent applicant: Nihon Shokuhin Kako Co., Ltd.

Claims (2)

[Claims] (1) A fatty liver inhibitor whose main component is hemicellulose obtained from corn bran or wheat bran. (2) The fatty liver inhibitor according to claim 1, which is obtained by removing starch, protein, etc. from corn bran or wheat bran and extracting the remainder with alkali.