JP2023077698A - Intestinal environment improver - Google Patents

Abstract

[PROBLEMS] To provide an intestinal environment-improving agent capable of enhancing the intestinal regulating action of Bifidobacterium to obtain a high effect. [Solution] Bifidobacterium longum and fructooligosaccharide or kestose are contained, and the bifidobacterium longum acts as a probiotic and the fructooligosaccharide or kestose acts as a prebiotic to produce a bifidobacterium. To obtain a remarkable intestinal regulation effect by explosively proliferating Bifidobacterium longum under the condition that single use of either Umlongum and fructo-oligosaccharide or kestose does not exhibit the function of proliferating bifidobacteria in the intestine. characterized by [Selection figure] None

Description

TECHNICAL FIELD The present invention relates to an intestinal environment-improving agent, and more particularly to an agent capable of enhancing the intestinal regulating action of Bifidobacterium to obtain a high effect.

The intestinal function means to regulate the condition of the stomach. The most familiar thing that can be experienced is to maintain a state of having regular bowel movements every day by improving bowel movements such as constipation and diarrhea. It can also be said that increasing useful bacteria such as Bifidobacterium and lactic acid bacteria existing in the intestine and reducing harmful bacteria such as Escherichia coli and Clostridium perfringens is one of the intestinal regulation effects. Overall, by increasing the number of useful bacteria in the intestines and increasing the amount of useful substances in the intestines and reducing harmful bacteria, the amount of putrefactive products is reduced, and as a result, the overall intestinal flora is balanced, which is beneficial to the body. can be defined as an intestinal regulation effect.

Among useful intestinal bacteria involved in such intestinal regulation effect, Bifidobacterium is a useful bacterium that exerts the greatest influence on the intestinal regulation effect. For example, diarrhea and constipation are typical examples of intestinal regulation effects associated with intestinal bifidobacteria. In addition, among the metabolites produced by intestinal bacteria, Bifidobacterium suppresses the growth of harmful bacteria in the intestine and prevents harmful bacteria from entering the body by producing highly bactericidal "acetic acid". It has an anti-infection effect that prevents the colonization of the virus. Furthermore, it is known that it is involved in the immunoregulatory action of the living body and exerts various effects such as stimulating intestinal immunity, enhancing immunity, protecting against infection, suppressing carcinogenesis, and alleviating allergic symptoms. In addition, it has been reported that bifidobacteria are reduced in the intestines of metabolic diseases represented by symptoms related to lifestyle-related diseases.

In other words, it can be said that how to increase the number of bifidobacteria in the intestine is the most important issue when exerting the intestinal regulation effect defined above. A representative method for increasing intestinal bifidobacteria is ingestion of probiotics/prebiotics.

Probiotics are defined as living microorganisms that act on bacteria present in the gastrointestinal tract or exert beneficial effects in living organisms when ingested alone. A wide range of items fall under that definition. Among them, Bifidobacterium is one of the probiotics that has been most frequently tried to be ingested in a live state as a probiotic. In particular, Bifidobacterium longum is a bacterium that is widely present in the intestines from infants to the elderly, and many research reports have been made as probiotics. For example, it is known that ingestion of Bifidobacterium longum as a probiotic increases intestinal bifidobacteria and regulates the intestines.
Prebiotics are defined as indigestible food ingredients that can selectively increase useful bacteria in the intestine, and typical examples thereof include dietary fiber and oligosaccharides. For example, it has been reported that fructooligosaccharide, which is one of the typical oligosaccharides, increases bifidobacteria in the intestine and provides an intestinal regulation effect by ingesting it for a specific period. Kestose, which is known as an active ingredient of fructo-oligosaccharides, is reported to proliferate bifidobacteria and reduce harmful bacteria.

From the above reports, it is suggested that the intake of bifidobacteria as probiotics or the intake of prebiotics is desirable as a means of increasing the number of bifidobacteria in the intestine. However, there is a problem in the proliferation of intestinal bifidobacteria by prebiotic intake.

For example, probiotics are a problem of intestinal colonization. Probiotics exist in the human intestine, but they are industrially developed live bacteria, so if they settle in the intestines for a long time, they may become pathogenic bacteria. Since it cannot eradicate the nature, it must be completely excreted from the body within a certain period of time (approximately 24 to 48 hours) without remaining in the body. That is, unless it is taken every day, the effect of regulating the intestines including the effect of increasing bifidobacteria cannot be exhibited.

In addition, since prebiotics selectively increase the bacteria present in the intestine, once the number of useful bacteria in the intestine increases, daily intake is not necessary like probiotics, but beneficial bacteria In order to increase Since the effective dose varies among individuals, beneficial bacteria including bifidobacteria in the intestine do not increase unless the effective dose is ingested.
In addition, in order to enhance the effects of probiotics/prebiotics, there is a method called synbiotics in which they are taken in combination. Synbiotics have been reported to be more effective than taking each alone.

BMC Microbiol. 2016 May 25;16:90. Doi: 10.1186/s12866-016-0708-5. Age-related changes in gut microbiota composition from newborn to centenarian: a cross-sectional studyToshitaka Odamaki, Kumiko Kato, Hirosuke Sugahar a, Nanami Hashikura, Sachiko Takahashi, Jin-Zhong Xiao, Fumiaki Abe, Ro Osawa J Pediatr Gastroenterol Nutr. 2016 Jul; 63 Suppl 1:S25-6. Probiotics for Irritable Bowl Syndrome: Clinical Data in Children. Eleonora Giannetti, Annamaria Staiano Effects of Bifidobacterium longum BB536 and Lactobacillus rhamnosus HN001 in IBS pa-tients. Bonfrate L, Di Palo DM, Celano G, Albert A, Vitellio P, De Angelis M, Gobbetti M, Portincasa p. Eur J Clin Invest. 2020 Mar;50(3):e13201. doi: 10.1111/eci. 13201. Epub 2020 Feb 12. Bifidobacterium mixture (B longum BB536, B infantis) M-63, B breve M-16V) treatment in children with seasonal allergic rhinitis and intermittent asthma. .iraglia Del Giudice M, Indolfi C, Capasso M, Maiello N, Decimo F, Ciprandi G.; . The capacity of short-chain fructooligosaccharides to stimulate facial bifidobacteria: a dose-response relationship study in healthy humans nutrition Journal 2006, 5:88 Bouhnik Y et al. . Yakult SCIENCE REPORT No. 10 Kanazawa, H.; et al. : Synbiotics reduce postoperative infectious complications: a randomized controlled trial in biliary cancer patients under- going hepatectomy. Langenbecks Arch. Surg. , 390, 104-113, (2005)) Kamiya, S.; et al. : The value of bill replacement during external biliary drainage: an analysis of intestinal performance, integrity, and microflora. Ann. Surg. , 239, 510-517, (2004)) Symbiotic Sugawara, G.; et al. : Perioperative synbiotic treatment postoperative infectious complications in biliary cancer surgery: a randomized controlled trial. Ann. Surg. , 244, 706-714, (2006) Kentaro Shimizu et al.: Intestinal flora, changes in the intestinal environment and efficacy of synbiotic therapy in patients with SIRS, Journal of Emergency Medicine, 17, 833-844, (2006) Shimizu, K, et al. : Altered gut flora and environment in patients with severe SIRS. J. Trauma. , 60, 126-133, (2006) Takashi Asahara et al.: Symbiotic therapy using Bifidobacterium and its mechanism of action, Milk industry technology, 58, 69-76 (2008) Kentaro Shimizu et al.: Intestinal therapy for critically ill patients: synbiotic therapy, ICU and CCU, 32, (2), 133-143, (2008) Shimizu, K.; et al. : Synbiotics decrease the incidence of septic complications in patients with severe SIRS: a preliminary report. Dig. Dis. Sci. , 54, 1071-1078, (2009) Japanese Patent Application Laid-Open No. 2005-306781

The above-described conventional configuration has the following problems.
Synbiotics can synergistically increase bifidobacteria, which is the main body of intestinal regulation effect, and under conditions where each of the above-mentioned probiotics / prebiotics does not exert its effect alone, bifidobacteria There is no prior art that examines whether it is possible to exert a proliferation effect.
In other words, it can be said that there is room for developing a synbiotics approach for exhibiting more effective bifidobacteria-proliferating abilities of probiotics and prebiotics.

The present invention has been made on the basis of these points, and its object is to provide an intestinal environment-improving agent. An object of the present invention is to provide an environmental improvement agent.

In order to achieve the above object, the intestinal environment improving agent according to claim 1 of the present invention is characterized by containing Bifidobacterium longum and fructo-oligosaccharide or kestose.
The intestinal environment-improving agent according to claim 2 is the intestinal environment-improving agent according to claim 1, which is characterized by being ingested once every 3 to 10 days.
The intestinal environment-improving agent according to claim 3 is the intestinal environment-improving agent according to claim 1 or claim 2, wherein the dose of kestose per 1 kg of body weight for an adult human is 27 mg or more. and a single dose of Bifidobacterium longum per 1 kg of body weight is 54.06 million cfu or more.

As described above, according to the intestinal environment improving agent according to claim 1 of the present invention, since it contains Bifidobacterium longum and fructo-oligosaccharides or kestose, the intestinal regulation action by Bifidobacterium is strengthened and a high effect is obtained. be able to.
Moreover, since the intestinal environment improving agent according to claim 2 is ingested once every 3 to 10 days in addition to the intestinal environment improving agent according to claim 1, the intestinal regulation effect can be obtained more effectively.
The intestinal environment-improving agent according to claim 3 is the intestinal environment-improving agent according to claim 1 or claim 2, wherein the dose of kestose per 1 kg of body weight for an adult human is 27 mg or more. In addition, since the amount of Bifidobacterium longum taken at one time per 1 kg of body weight is 54,060,000 cfu or more, a more effective intestinal regulation effect can be obtained.

An embodiment of the present invention will be described below.
The intestinal environment improving agent according to this embodiment uses Bifidobacterium long gum as a probiotic and fructooligosaccharide or kestose as a prebiotic, so that Bifidobacterium long gum and fructooligosaccharide or Bifidobacterium longum proliferates explosively under conditions where single use of any kestose does not exhibit the function of proliferating bifidobacteria in the intestine, and a remarkable intestinal regulation effect is obtained.

In addition, by ingesting the intestinal environment-improving agent once every 3 to 10 days, a remarkable intestinal regulation effect can be obtained.
In the intestinal environment-improving agent of the present invention, for an adult human, the dose of kestose per 1 kg of body weight is 27 mg or more, and the dose of Bifidobacterium longum is 27 mg or more per 1 kg of body weight. is 54,060,000 cfu or more, a high effect can be obtained.

In addition, fructooligosaccharides and kestose are ingredients that are blended in commonly distributed cosmetics, health foods, and general foods, so they are safe and highly effective.
In addition, it can be used simply by mixing it with meals, and the method of use is simple, so that treatment can be easily continued.

Hereinafter, this one embodiment will be specifically described with reference to examples.
However, the present invention is not limited to these examples.

[Example 1]
In this Example 1, experiments were performed using 8-week-old male Sprague-Dawley rats.
First, 5×9=45 8-week-old male Sprague-Dawley rats were obtained. The source of the above rats is Japan SLC (Hamamatsu, Japan).
The rats are then kept at a predetermined temperature (23±1° C.) with a wire mesh gauge placed in an animal room that is illuminated for 12 hours a day and dark the rest of the day. The rats were housed one per person in a wire mesh cage.
The rats were then acclimated within the wire mesh cage for one week.

The rats were then divided into groups of 5 rats, and the first group was fed a diet mixed with saline and the second group received 200 mg fructooligosaccharides (mayoligosaccharides) daily for one month. P, manufactured by Meiji Co., Ltd., the same applies hereinafter), and the third group was fed with 200 mg of kestose (Kestose 95, manufactured by Bussan Food Science Co., Ltd., the same applies hereinafter). The group was given feed mixed with 1 billion cfu of Bifidobacterium longum 1 (Bifidobacterium longum, manufactured by Seti Co., Ltd., hereinafter the same), and the fifth group was fed with 1 billion cfu of Bifidobacterium longum 2. (BB536, manufactured by Morinaga Milk Industry Co., Ltd., the same applies hereinafter), the sixth group was given a feed mixed with 1 billion cfu of Bifidobacterium longum 1 and 200 mg of fructo-oligosaccharides, and the seventh group group was fed with 1 billion cfu of Bifidobacterium longum 2 and 200 mg of fructo-oligosaccharides, and the eighth group was fed with 1 billion cfu of Bifidobacterium longum 1 and 200 mg of kestose. A ninth group was fed a diet mixed with 1 billion cfu of Bifidobacterium longum 2 and 200 mg of kestose.

Thereafter, the cecal contents were rapidly collected from the rats under anesthesia with isoflurane, frozen in liquid nitrogen and stored at −80° C., then genomic DNA was extracted from the cecal contents and quantitative real-time PCR (qPCR) was performed. ) to quantify the number of bifidobacteria.
Genomic DNA extraction from cecal contents and quantitative real-time PCR (qPCR) to quantify the number of bifidobacteria were performed by the following procedure. That is, after performing crushing treatment by using a bead crusher "FastPrepFP100A" (MPBiomedical), a nucleic acid extractor "Magtration (registered trademark) System 12GC" and its dedicated reagent "MagDEA (registered trademark) DNA200" (Precision System・Science Co.) was used to extract genomic DNA according to the manufacturer's instructions. Using this genomic DNA as a template, Bifidobacterium longumsubsp. Using primers (described below) specific to 16S rDNA of longum JCM1217, real-time PCR reagent "SYBR (registered trademark) PremixExTaqII (TliRNaseHPlus)" (Takara) and real-time PCR device "Rotor-GeneQ" (QIAGEN) Real-time PCR was performed to determine the number of 16S rDNA copies of each microorganism per gram of cecal contents.
The real-time PCR is
Forward primer to GATTCTGGCTCAGGATGAACGC
year,
Revers primer to CTGATAGGACGCGACCCCAT
year,
The program conditions were 35 cycles of 95°C (5 sec)-60°C (20 sec)-72°C (20 sec).
Table 1 shows the results.

In Table 1 above, Bifidobacterium longumsubsp. The 16S rDNA copy number of longum JCM 1217 was used as an index, and the value was described as log10. There are three types of each value: median (50% IQR), 25% IQR, and 75% IQR. Statistical analysis was performed using Microsoft Excel's Student's t-test, and P-values were considered significant at <0.05 compared to the first group of mixed saline and food.
According to Table 1, compared with the first to fifth groups, the number of bifidobacteria is clearly increased in the sixth to ninth groups in which fructooligosaccharides or kestose and Bifidobacterium longum are mixed in the feed.
Therefore, it can be said that it is better to mix Bifidobacterium longum with fructooligosaccharides or kestose in the feed.

[Example 2]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats, and the 10th group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum 1 daily for 1 month. Groups were fed a diet of 200 mg kestose and 5 billion cfu Bifidobacterium longum 1 and a 12th group was fed a diet of 200 mg kestose and 100 million cfu Bifidobacterium longum 1. and a thirteenth group was fed a diet containing 200 mg of kestose and 0.1 billion cfu of Bifidobacterium longum 1 .
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 2 as in Example 1 above.

According to Table 2, the number of Bifidobacterium longum was detected to be large in both groups. In the 11th group containing gum 1 and the 12th group containing 100 million cfu of Bifidobacterium longum 1, the number of bifidobacteria was particularly large, and 10 billion cfu of Bifidobacterium longum was detected in the 12th group. The tenth group containing Gum 1 had the highest number of bifidobacteria detected.
Therefore, it can be said that Bifidobacterium longum 1 should be mixed with 100 million cfu or more.
When converted to one intake per kg of body weight in rats, 100 million cfu/0.3 kg = 333,333,333 cfu/kg. (Rats weigh 300 g.)
In addition, one intake per kg body weight in rats can be converted to one intake per kg body weight in adult humans by Equation 1. (Shannon Reagan-Shaw et al., The FASEB Journal, 40 Vol. 22, March 2007, pp. 659-661)
Y = X × 6 ÷ 37 (Formula 1)
However, Y: one dose per 1 kg body weight of an adult human
X: one-time intake per 1 kg body weight in rats Converting one-time intake per 1 kg body weight in rats to one-time intake per 1 kg body weight in an adult human is 333,333,333 cfu/kg × 6 ÷ 37 = 54,054,054 cfu/kg or more.

[Example 3]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and the 14th group was fed a diet containing 500 mg of kestose and 1 billion cfu of Bifidobacterium longum 1 daily for 1 month. Groups were fed a diet containing 100 mg kestose and 1 billion cfu Bifidobacterium longum 1, and a sixteenth group was fed a diet containing 50 mg kestose and 1 billion cfu Bifidobacterium longum 1. and a seventeenth group was fed a diet containing 10 mg of kestose and 1 billion cfu of Bifidobacterium longum 1.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 3 as in Example 1 above.

According to Table 3, a large number of bifidobacteria was detected in all of them. A particularly large number of bifidobacteria was detected in 16 groups, and the largest number of bifidobacteria was detected in the 14th group containing 500 mg of kestose.
Therefore, it can be said that it is good to mix 50 mg or more of kestose.
When converted to one intake per kg body weight in rats, 50 mg/0.3 kg = 166.7 mg/kg. (Rats weigh 300 g.)
Moreover, when this is converted into one intake per kg body weight of an adult human, it is 166.7 mg/kg x 6/37 = 27 mg/kg or more.

[Example 4]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and the 18th group was fed saline-mixed diet and the 19th group was fed 200 mg once every 3 days for one month. The 20th group was fed a diet mixed with fructooligosaccharides, the 20th group was fed a diet mixed with 200 mg kestose, the 21st group was fed a diet mixed with 10 billion cfu Bifidobacterium longum 1, Group 22 was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 2 and group 23 was fed a diet mixed with 200 mg fructo-oligosaccharides and 10 billion cfu of Bifidobacterium longum 1. The 24th group was fed with 200 mg of fructooligosaccharide and 10 billion cfu of Bifidobacterium longum 2, and the 25th group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum. The 26th group was fed a diet mixed with gum 1 and the 26th group was fed a diet mixed with 200 mg kestose and 10 billion cfu Bifidobacterium longum 2.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 4 in the same manner as in Example 1 above.

According to Table 4, the 19th and 20th groups fed with a diet mixed with only fructo-oligosaccharides or kestose, and the 21st group fed with a diet mixed with only Bifidobacterium longum 1 or Bifidobacterium longum 2 , in group 22, no higher numbers of bifidobacteria were detected than in group 18, which was fed a diet mixed with physiological saline; More bifidobacteria counts were detected in the 23rd to 26th groups fed with the diet mixed with Umron Gum 2 compared to the 18th group.

[Example 5]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and the 27th group was fed saline-mixed diet and the 28th group was fed with 200 mg once every 5 days for 1 month. The 29th group was fed a diet mixed with fructooligosaccharides, the 29th group was fed a diet mixed with 200 mg kestose, the 30th group was fed a diet mixed with 10 billion cfu Bifidobacterium longum 1, Group 31 was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 2 and Group 32 was fed a diet mixed with 200 mg fructo-oligosaccharides and 10 billion cfu of Bifidobacterium longum 1. The 33rd group was fed with 200 mg of fructooligosaccharide and 10 billion cfu of Bifidobacterium longum 2, and the 34th group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum. The 35th group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum 2.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 5 as in Example 1 above.

According to Table 5, the 28th and 29th groups fed with a diet mixed with only fructo-oligosaccharides or kestose, and the 30th group fed with a diet mixed with only Bifidobacterium longum 1 or Bifidobacterium longum 2 , compared to the 31st group, the 32nd to 35th groups fed with fructooligosaccharides or kestose mixed with either Bifidobacterium longum 1 or Bifidobacterium longum 2 had a higher Bifidobacterium count. was detected. However, it is less than in Example 4, which was given once every three days.
No effective bifidobacteria count was detected in the 27th group fed with the saline-mixed feed.

[Example 6]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and once every 10 days for a month, the 36th group was fed saline mixed diet, and the 37th group was given 200 mg The 38th group was fed a diet mixed with 200 mg of kestose, the 39th group was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 1, Group 40 was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 2, and Group 41 was fed a diet mixed with 200 mg fructo-oligosaccharides and 10 billion cfu of Bifidobacterium longum 1. The 42nd group was fed with 200 mg of fructooligosaccharide and 10 billion cfu of Bifidobacterium longum 2, and the 43rd group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum. The 44th group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum 2.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 6 as in Example 1 above.

According to Table 6, the 36th group given a diet mixed with physiological saline, the 37th and 38th groups given a diet mixed only fructo-oligosaccharides or kestose, and Bifidobacterium longum 1 or Bifidobacterium Bifidobacterium was not detected in the 39th and 40th groups that were given feed mixed with Umlongum 2 only, but both fructo-oligosaccharides or kestose and Bifidobacterium longum 1 or Bifidobacterium longum 2 were fed. High bifidobacteria counts were detected in groups 41 to 44 fed mixed feed. However, it is less than in Example 5, which was given once every 5 days.

[Example 7]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and once every 15 days for a month, the 45th group was fed saline-mixed diet, and the 46th group was fed with 200 mg. The 47th group was fed a diet mixed with 200 mg of kestose, the 48th group was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 1, Group 49 was fed a diet mixed with 10 billion cfu of Bifidobacterium longum 2, and group 50 was fed a diet mixed with 200 mg fructo-oligosaccharides and 10 billion cfu of Bifidobacterium longum 1. The 51st group was fed with 200 mg of fructooligosaccharide and 10 billion cfu of Bifidobacterium longum 2, and the 52nd group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum. The 53rd group was fed with 200 mg of kestose and 10 billion cfu of Bifidobacterium longum 2.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 7 as in Example 1 above.

According to Table 7, the 45th group given a diet mixed with physiological saline, the 46th and 47th groups given a diet mixed only fructo-oligosaccharides or kestose, and Bifidobacterium longum 1 or Bifidobacterium Bifidobacterium was not detected in the 48th and 49th groups that were given feed mixed only with Umlongum 2, but both fructo-oligosaccharides or kestose and Bifidobacterium longum 1 or Bifidobacterium longum 2 were fed. Bifidobacterium counts were detected in the 50th to 53rd groups fed the mixed feed. However, it was less than in Example 6, which was given once every 10 days, and was about the same as the 18th group, which was given the feed mixed with physiological saline once every 3 days.
Therefore, from the results of Examples 4 to 7, it can be said that the agent for improving the intestinal environment according to the present invention should be given at least once every 10 days.

[Example 8]
Rats were prepared as in Example 1.
The above rats were divided into groups of 5 rats, once every 3 days for 1 month, and the 54th group was fed with 200 mg of kestose and 10 billion of Bifidobacterium longum 1. The 55th group was given a diet mixed with 200 mg of kestose and 5 billion cfu of Bifidobacterium longum 1, and the 56th group was given 200 mg of kestose and 1 billion cfu of Bifidobacterium longum. The 57th group was fed a diet mixed with 200 mg kestose and 100 million cfu Bifidobacterium longum 1, the 58th group was fed a diet mixed with 200 mg kestose and 0.1 Feed mixed with 100 million cfu of Bifidobacterium longum 1 was given.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 8 as in Example 1 above.

According to Table 8, the 54th group fed with 200 mg of kestose mixed with 10 billion cfu of Bifidobacterium longum 1, the 54th group fed with 200 mg of kestose mixed with 5 billion cfu of Bifidobacterium longum 1. Fifty-fifth group fed, 200 mg kestose mixed with 1 billion cfu Bifidobacterium longum 1 Fifty-sixth group fed 200 mg kestose and 100 million cfu Bifidobacterium longum 1 Good results were obtained in the 57th group fed with Bifidobacterium longum 1, but in the 58th group fed with 200 mg of kestose and 0.1 billion cfu of Bifidobacterium longum 1. No valid results were obtained.
Therefore, when giving once every three days, it can be said that the intestinal environment improving agent according to the present invention containing 100 million cfu or more of Bifidobacterium longum 1 should be given.
Converting to one intake per kg body weight in rats, 100 million cfu/0.3 kg = 333,333,333 cfu/kg. (Rats weigh 300 g.)
Moreover, when this is converted into one-time ingestion per 1 kg body weight of an adult human, it is 333,333,333 cfu/kg×6÷37=54,054,054 cfu/kg or more.

[Example 9]
Rats were prepared as in Example 1.
The rats were divided into groups of 5 rats each, and once every 3 days for 1 month, the 59th group was mixed with 500 mg of kestose and 1 billion cfu of Bifidobacterium longum 1. The 60th group was fed with 200 mg of kestose and 1 billion cfu of Bifidobacterium longum 1, and the 61st group was fed with 100 mg of kestose and 1 billion cfu of Bifidobacterium longum. The 62nd group was fed a diet mixed with 50 mg kestose and 1 billion cfu Bifidobacterium longum 1, the 63rd group was fed a diet mixed with 10 mg kestose and 1 billion cfu. Feed mixed with cfu of Bifidobacterium longum 1 was given.
Thereafter, in the same manner as in Example 1, genomic DNA was extracted from the cecal contents of the rats, and the number of bifidobacteria was quantified by quantitative real-time PCR (qPCR).
The results are shown in Table 9 as in Example 1 above.

According to Table 9, the 59th group fed with 500 mg of kestose and 1 billion cfu of Bifidobacterium longum 1, the 59th group with 200 mg of kestose and 1 billion cfu of Bifidobacterium longum 1; A 60th group fed with a diet consisting of 100 mg kestose and 1 billion cfu Bifidobacterium longum 1 A 61st group fed with 50 mg kestose and 1 billion cfu Bifidobacterium longum 1 Good results were obtained in the 62nd group fed with Bifidobacterium longum 1, while the 63rd group fed with 10 mg kestose and 1 billion cfu of Bifidobacterium longum 1 showed no effect. No results were obtained.
Therefore, when giving once every three days, it is preferable to give the intestinal environment-improving agent of the present invention containing 50 mg or more of kestose.
When converted to one intake per kg body weight in rats, 50 mg/0.3 kg = 166.7 mg/kg. (Rats weigh 300 g.)
Moreover, when this is converted into one intake per kg body weight of an adult human, it is 166.7 mg/kg x 6 waru 37 = 27 mg/kg or more.

[Example 10]
Women in their thirties were divided into groups of three, once a day for two months, the first group took only 3g of fructo-oligosaccharides and the second group took only 3g of kestose. The third group received 1 billion cfu of Bifidobacterium longum 1 only, and the fourth group received 3 g of fructo-oligosaccharides and 1 billion cfu of Bifidobacterium longum 1 per dose. A fifth group received a mixture of 3 g of kestose and 1 billion cfu of Bifidobacterium longum 1 per serving.
Assuming that the body weight is 50 kg, 60 mg of fructo-oligosaccharide or kestose and 20 million cfu of Bifidobacterium longum 1 were taken per 1 kg of body weight.

The following items were scored, and the evaluation was performed based on the average value of all the items, and the average value was calculated for each group.
(1) Smell of stools has disappeared (2) Frequent use has increased (3) Feeling refreshed (4) Shape has improved (5) Soft stools have decreased (6) Overall stomach condition felt better

The scores for each of the above items were evaluated according to the following criteria.
4 points: Very strongly felt 3 points: Strongly felt 2 points: Somewhat felt 1 point: Slightly felt 0 points: Not felt at all Table 10 shows the results.

According to Table 10, the 9th group ingested 1 billion cfu of Bifidobacterium longum 1 with 3 g of fructooligosaccharides, the 10th group ingested 1 billion cfu of Bifidobacterium longum 1 with 3 g of kestose. Good results were obtained in the group, but no effective results were obtained in the 6th, 7th and 8th groups ingesting fructo-oligosaccharides, kestose and Bifidobacterium longum 1 individually.

[Example 11]
Women in their thirties were divided into groups of three, and once every three days for two months, the sixth group took only 3g of fructooligosaccharides and the seventh group took only 3g of kestose. The eighth group took only 1 billion cfu of Bifidobacterium longum 1 per dose, and the ninth group mixed 3g of fructo-oligosaccharides and 1 billion cfu of Bifidobacterium longum 1 per dose. and the tenth group mixed 3 g of kestose and 1 billion cfu of Bifidobacterium longum 1 per dose.
Evaluation was performed in the same manner as in Example 10 described above.
Table 11 shows the results.

According to Table 11, the 9th group ingested 1 billion cfu of Bifidobacterium longum 1 with 3 g of fructooligosaccharides, the 10th group ingested 1 billion cfu of Bifidobacterium longum 1 with 3 g of kestose. Good results were obtained in the group, but no effective results were obtained in the 6th, 7th and 8th groups ingesting fructo-oligosaccharides, kestose and Bifidobacterium longum 1 individually.
In addition, even if it was ingested once every three days, the results were comparable to those in Example 10 in which it was ingested once every day.

Thus, the intestinal environment-improving agent containing fructo-oligosaccharide can enhance the intestinal regulating action of bifidobacteria and obtain a high effect. In addition, since fructooligosaccharides and kestose are ingredients contained in commonly distributed cosmetics, health foods, and general foods, they can effectively strengthen the intestinal function without side effects.

TECHNICAL FIELD The present invention relates to an agent for improving the intestinal environment, and more particularly to an agent devised to enhance the intestinal regulation action of Bifidobacterium to obtain a high effect. is.

Claims (3)

An intestinal environment-improving agent comprising Bifidobacterium longum and fructo-oligosaccharide or kestose. In the intestinal environment improving agent according to claim 1,
An intestinal environment-improving agent characterized by being used so as to be ingested once every 3 to 10 days. In the intestinal environment improving agent according to claim 1 or claim 2,
For human adults,
A single intake of kestose is 27 mg or more per kilogram of body weight;
A dose of Bifidobacterium longum per 1 kg of body weight is 54.06 million cfu or more,
An intestinal environment improving agent characterized by being used in.