WO2004071520A1 - Preparation de bifidobacterium pour ameliorer les fonctions immunes - Google Patents

Preparation de bifidobacterium pour ameliorer les fonctions immunes Download PDF

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Publication number
WO2004071520A1
WO2004071520A1 PCT/JP2004/001683 JP2004001683W WO2004071520A1 WO 2004071520 A1 WO2004071520 A1 WO 2004071520A1 JP 2004001683 W JP2004001683 W JP 2004001683W WO 2004071520 A1 WO2004071520 A1 WO 2004071520A1
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WO
WIPO (PCT)
Prior art keywords
stress
fraction
bifidobacteria
preparation
bifidobacterial
Prior art date
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PCT/JP2004/001683
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English (en)
Japanese (ja)
Inventor
Akira Hosono
Hiroki Kobo
Ryo Nakamura
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The Food Science Institute Foundation
Nihon University
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Publication of WO2004071520A1 publication Critical patent/WO2004071520A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to a bifidobacterial preparation that improves the deterioration of the intestinal immune system of animals (including humans) induced by stress.
  • the bifidobacterial preparation of the present invention comprises administering a nutritional product containing a bifidobacterial preparation to an animal (including a human) before, during and / or after a stress event.
  • the present invention also relates to a nutritional product comprising an antioxidant system and a bifidobacterial preparation.
  • stress or stress response a common group of biological reactions to various external harmful factors (stressors), that is, medically physiologically described as general adaptation syndrome, is called stress or stress response. is there.
  • stress a common group of biological reactions to various external harmful factors (stressors), that is, medically physiologically described as general adaptation syndrome, is called stress or stress response.
  • stress Today, however, it is common to see changes in the body due to external factors, including social psychological factors, as “stress”.
  • stress one of the three symptoms of Se lye who advocated the concept of stress was the thymus' lymphatic atrophy. Causes quantitative and qualitative changes to the immune system.
  • the mechanism by which stress causes the quantitative and qualitative changes described above in an individual's immune system has been analyzed in a field called “psycho-neuroimmunology” or “neuroendocrine immunology”.
  • the immune system as a body defense system interacts with the nervous system and the endocrine system, and works to maintain homeostasis through the cross of the so-called “nerve-endocrine-one immune system”. Is becoming clear.
  • the intestinal tract immune system is characterized by IgA secretion to the mucosal surface and plays an important role in infection protection, but the effects of various stresses on this IgA secretion have not been clarified so far.
  • enterobacteria-derived components expected as probiotics have an intestinal immunity-modulating action (eg, Non-Patent Document 3).
  • Bifidobacteriun ⁇ Lactobacillus Some Gram-positive bacteria have functions that increase lymphocyte proliferation activity and IgA production, and are expected to have immunomodulatory effects of probiotics. Anti-tumor effects and adjuvant activities have been reported for Gram-positive bacterial cell wall components such as peptide daricans, but water-soluble high-molecular-weight polysaccharides among ⁇ /// cer / OT cell-derived components It has been reported that the ingredients also have high immunostimulatory action.
  • This active ingredient contains high-molecular polysaccharides composed mainly of glucose and galactose, and the constituent sugars are -4Galp- (or -5Gal i l-), _6Glcp ⁇ , Gal f l_,- Does not contain galactofuranoside such as 6Gal f l-
  • -4Galp- or -5Gal i l-
  • _6Glcp ⁇ Gal f l_
  • Gal f l_ Does not contain galactofuranoside such as 6Gal f l-
  • Non-Patent Document 1 Keller, SE et al: Science. 221: 1301-1303, 1983; Ke Her, SE et al: Proc. Natl. Acad. Sci. USA. 85: 9297-9301, 1988
  • Non-Patent Document 2 Hof iman-Goeiz, L. et al .: I ⁇ unol. Today. 15: 382-387, 1994
  • Non-Patent Document 3 Hosono m: FOOD Style 21 2002. 9 (Vol. 6 No. 9) Disclosure of Invention
  • the present invention provides a bifidobacterial preparation having an action of improving immunity resulting from stress in animals including humans, in particular, a decrease in function of the intestinal tract immune system.
  • the present invention also provides a bifidobacterial preparation having an action of imparting stress resistance to animals including humans. Stress includes excessive physical activity, extreme mental fatigue, and disease state.
  • the present invention was made on the basis of the finding that // ingestion of bacterial components of A3 ⁇ 4 bacteria can improve the functional deterioration of intestinal immunity induced by stress. is there.
  • a bifidobacteria preparation that improves stress resistance and Z or stress-induced dysfunction caused by microbial cells of genus / 7'ffl7,
  • fraction is a cell wall fraction, a cytoplasmic membrane fraction, a protoplast, or a water-soluble fraction
  • BifidobacteriuEM (hereinafter also referred to as “Bifidobacteria”) was studied by Reutei :, Mitsuoka, Scardovi and others, and was classified into 11 species in the 8th edition of Bergey's Manual. Later, classification by genotype using DNA homology was introduced by Scardovi, and many new bacterial species were recognized based on DNA homology. Berg According to the ey's Manual of Systematic Bacter iology (1986), it is classified into 24 species, and 26 species are currently recognized as Bifidobacteriun as shown in Table 1. 1992. Classification and ecology of intestinal flora: p. 55-72.
  • B. bifidum 58 Human infant and adult feces; human vagina; lactating calf feces
  • B. gallicum 61 Human feces also, ⁇ . Adolescent is, Manni] ⁇ 1 and sorbi! ⁇ 1 was divided into four biovars a, b, c, and d depending on the assimilation property. While B. catenulatim, B. angulatunk B. dentium, and Reported as a new strain isolated from human feces, vagina and oral cavity, sewage and calf. As shown in Table 2, these four strains are distinctly different from those of ⁇ zo c / in terms of DNA homology, but their glycolytic properties are considered to be similar (Tomookatsu Mitsuoka) Edit. 199 2. Classification and ecology of intestinal flora: p. 55-72.
  • candidate genus of 77 genera includes all of the bacterial species (live and dead) shown in Table 1 and Table 2. Bifidobacteria culture methods are known
  • bifidobacteria cultures also included are bifidobacteria cultures, purified bifidobacteria cells, freeze-dried cells, or commercially available bifidobacteria preparations. As for bifidobacteria preparations, there are many products that claim 1 X 10 1D Zg or more.
  • the present invention provides a crushed cell body fraction, a cell wall fraction, a pro-plus cocoon, a cytoplasmic membrane fraction, or a water-soluble fraction, and when subjected to various stresses, IgA antibodies in the intestinal mucosal system Including all those having the effect of reducing IgA production and / or mitigation of mitogenic activity on lymphocytes in / or cytokine production.
  • the cell-insoluble fraction containing the soluble cytoplasmic fraction of bifidobacteria and other membrane fractions can be easily separated by ultracentrifugation after sonication of the cells.
  • Preparation of cell wall fractions, protoplasts, cytoplasmic membrane fractions, or water-soluble fractions is well known [for example, Hiroshi Matsuzawa, Suiharu Lee, “Microbial Experiments (Neurochemistry Laboratory)”, edited by Japan Biochemical Society, ( Tokyo Chemical Co., Ltd., p. 179; Japanese Patent No. 3174611; Japanese Unexamined Patent Publication No. 9-241179], and the present invention includes these known techniques.
  • Bifidobacteria are distributed in the digestive tract and faeces of various human animals, the vagina and oral cavity of bees, the digestive tract of bees, and sewage, and it is "sumi" between humans and animal species and between infants and adults. Is recognized. At present, 10 species of bifidobacteria derived from humans have been reported, but in Japan, bifidum, B. infant is, B. breve, B. longum, B. adolescent is 5 species, and other species have not been reported (edited by Tomohiro Mitsuoka. 1992. Classification and ecology of intestinal flora: p. 55-72. Academic Publishing Center).
  • the candidate bifidobacteria are the above-mentioned 10 species resident in chicks, namely B. bifidu m, B. infant is, B. breve, B. long ⁇ B. adolescent is, B. angulatum, B. ca teniilati B. pseudoca tenula turn, B. dentiun ⁇ B. zozo.
  • the target is a domestic cattle
  • the bifidobacteria derived from cattle in Table 1, i.e. B. bifiduE, B. adolescent is, B. pseudoca tenula turn, B. globosum, B.
  • B. animal is ⁇ B. thermophilum and X B. ffi ⁇ If the subject is a chicken, the same idea is applied to B. pseudolong belly, B. animal is, B. thermop. hilum, B. pullorm ⁇ XM ⁇ zo / ⁇ ⁇ are candidates. In addition, bifidobacteria isolated from nature will be candidates.
  • Bifidobacteria of microorganism preservation organizations include: ⁇ . Zo ATCC 15707 T (Intes tin of adult), B. (Intes tine of infant), B. Man ti ski CC 15697 T strain (Intes tine of infant), B. (Intestine of adult), caienuJanAmi (Int. J. Syst. Bact. 24: 6-20, 1974) and the like.
  • the present invention includes such a combination.
  • the preparation containing the disrupted bifidobacteria, cell wall fraction, protoplast, cytoplasmic membrane fraction, or water-soluble fraction of the present invention is administered to animals containing baboons whose immune function has been reduced by stress for the purpose of improving immune function.
  • animals containing baboons whose immune function has been reduced by stress can also be used as a health functional food for humans whose immune function has been reduced by stress.
  • an effective amount thereof can be added to the feed for the purpose of improving immune function.
  • Antibiotics that exert an advantageous effect on immune function such as vitamin A, selenium, vitamin C or] 3-carotene can be added to the bifidobacteria preparation.
  • the effective amount of a bifidobacteria preparation for humans whose immune function has been reduced due to stress is to improve immune function or improve immune function is empirically more than 10 9 viable bacteria per day Since the intestinal regulating effect is recognized in this study, this number is considered to be a standard (Ogata T. et al .: Bioscience Microflora, 16 (2): 5 3-58, 1997).
  • An effective amount of improvement can be determined from known immunological parameters (eg, activation of leukocyte function).
  • the form of the bifidobacteria preparation of the present invention can be, for example, a lump, liquid, syrup, or powder.
  • various additives such as a bulking agent, a sweetener, and other sugars It can contain quality, vitamins, fragrances, coloring agents, etc.
  • Forms include liquid or powder food forms, and pharmaceutical forms such as powders, powders, liquids, suspensions, tablets, and foaming agents.
  • FIG. 1 is a diagram showing the measurement results of blood corticosterone during exercise stress, restraint stress, water invasion restraint stress, and non-stress loading period in BALB / c mice.
  • exercise stress image
  • restraint stress ⁇
  • water invasion restraint stress ⁇
  • non-stress X.
  • Fig. 2 shows the total measurement results in the intestinal contents after 5 days of stress applied to BALB / c mice.
  • Fig. 3 is a diagram showing the results of measuring Peyer's patch cells in the presence of Con A or BP and measuring IgA in the culture supernatant after loading the above-mentioned strains on BALB / c mice for 5 days.
  • Fig. 4 shows that Bifidobacterium pseudocatenulatum l ⁇ W ⁇ (BP) was orally administered at 10 mg / day for 5 days, after restraint stress (5 hours per day) was loaded for 5 consecutive days, and then in the intestinal contents It is a figure which shows the result of having measured the total IgA amount.
  • FIG. 5 is a diagram showing the results of measuring the IFN-a in the culture supernatant of the same Peyer's patch cells cultured in the presence of Con A or BP.
  • Bifidobacterium pseudocatenulatum 7041 (Japan Bifidobacterium Center 1) 3 Anaerobic culture was performed at 7 ° C for 48 hours.
  • the cells are collected by centrifugation (6,000 X g, 5 ° C, 10 minutes), washed 3 times with ultrapure water (6,000 X g, 5 ° C, 10 minutes), and then frozen. Dried and stored at 20 ° C. Freeze-dried cells are suspended in ultrapure water at 4 mg / mL, then sonicated under ice-cooling (Otake, 15 minutes), and further centrifuged at 800X g, 5 ° C for 5 minutes. Metal scraps generated by the treated cells and sonication were removed. This was freeze-dried to obtain cell debris (BP) (BP was stored at -20 ° C until Atsey).
  • BP cell debris
  • mice Female BALB / c mice were purchased at 6 weeks of age (Claire Japan), acclimated for 3 days, and then divided into 4 groups, 8 per group. Next, the exercise stress group only was trained for 5 days from the 4th to the 8th day, with running water swimming exercise (water temperature 32 ° C, 7 L / min for 5 minutes x 3 sets). Each group was stressed under the following conditions for 5 days from 11 to 15 days. During the experimental period (15 days), each mouse was given free access to solid food MF (oriental yeast) for mice and water.
  • solid food MF orientation yeast
  • the stress conditions are exercise stress, water temperature 32 ° C, 7L / miri running water swimming exercise load (10 minX3 set per day), restraint stress (in a 50 mL polypropylene tube with many small holes) Mouse is fixed) Load group (3 hours a day), water invasion restraint stress (Mouse is fixed in a 50 mL polypropylene tube with many small holes, and this is 1 cm in depth, approx. 32 ° C) It was placed in a cage containing a large amount of water). It was classified as a load group (3 hours a day) and a non-stress load group.
  • Blood was collected from the orbital venous plexus for 5 days before stress was applied (Day 10) and during the stress period (Days 11, 12, 13, 14, and 15), and the amount of corticosterone in the plasma was measured using the RIA method. More measured.
  • Goat anti-mouse IgA antibody (ICN Pharmaceuticals) was added to a 96-well microphone mouthplate (MaxSorp, Nunc) and immobilized at 4 ° C overnight. After blocking with urine serum albumin (BSA), a sample diluted with 1% BSA-PBS-Tween solution (sample buffer) to the appropriate magnification was added to each well, and incubated at 4 ° C. . Piotinylated anti-mouse IgA antibody (Sigma) diluted 1,000-fold with sample buffer was added to each well and incubated at room temperature for 2 hours.
  • BSA-PBS-Tween solution sample buffer
  • Streptavidin-alkaline phosphatase complex (Zymed) diluted 1,000-fold with 1% BSA-PBS-Tween solution was added to each well and incubated at room temperature for 1 hour. Then, 0.1% phenyltriphosphate ninatrium-containing diethanolamine buffer was added to each well, incubated at 37 ° C, and the absorbance was measured at 405 nm (BioRad). The total IgA content in the intestinal contents was also measured by the same method.
  • the total IgA content in the intestinal contents was significantly lower in the exercise stress group than in the non-stress group (control group) by 0.05, one-way analysis of variance and Tukey's test.
  • the restraint stress and water invasion restraint stress load groups tended to show lower values than the unstressed group (Fig. 2).
  • mice Female BALB / c mice were purchased at 6 weeks of age (CLEA Japan) and acclimated for 3 days, then 8 animals per group: 1) restraint stress load + BP administration, 2) restraint stress load + BP not applied 3) Restraint stress unloaded + BP administration, and 4) Restraint stress unloaded + BP non-administered.
  • the total amount of IgA in the intestinal contents tended to be higher in the BP administration group than in the BP non-administration group in the restraint stress load group (Fig. 4).
  • the total IgA level in the BP non-administered group in the restraint stress load group was almost the same as the total IgA level in the BP-administered group and the non-BP-administered group in the non-stressed group.
  • the amount of IFN- ⁇ produced by Peyer's patch cells was remarkably increased by administering BP to the restraint stress and non-restraint stress groups.
  • a bifidobacteria preparation useful for improving the functional deterioration of the intestinal tract immune system resulting from stress is provided.
  • the preparation is useful as a health functional food. Stress includes excessive physical exercise, mental extreme fatigue, and disease states.
  • Stress includes excessive physical exercise, mental extreme fatigue, and disease states.
  • a bifidobacterial preparation having the formula:
  • a bifidobacteria preparation having an effect of imparting stress resistance to animals including humans has been provided. Stress includes excessive physical exercise, extreme mental fatigue, and disease state.

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  • Health & Medical Sciences (AREA)
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  • Veterinary Medicine (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
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  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

Il a été découvert que l'administration par voie orale de constituants de cellules de Bifidobacterium contribue à l'amélioration de l'hypofonction induite par stress de l'immunité dans le tractus intestinal.
PCT/JP2004/001683 2003-02-17 2004-02-17 Preparation de bifidobacterium pour ameliorer les fonctions immunes WO2004071520A1 (fr)

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JP2003038831A JP2004262773A (ja) 2003-02-17 2003-02-17 免疫機能改善を目的としたビフィズス菌製剤
JP2003-038831 2003-02-17

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Cited By (1)

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US10028981B2 (en) 2009-08-25 2018-07-24 Nestec S.A. Bifidobacterium longum and functional GI disorders

Families Citing this family (6)

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WO2006087913A1 (fr) * 2005-02-02 2006-08-24 Meiji Dairies Corporation Composition pour l'activation immunitaire
WO2007122885A1 (fr) 2006-03-31 2007-11-01 Morinaga Milk Industry Co., Ltd. Régulateur de production d'interleukine, composition pharmaceutique ou aliment le comprenant et sa méthode de production
JP2008195635A (ja) * 2007-02-09 2008-08-28 Crossfield Bio Inc 馬用乳酸菌製剤
JP2009100692A (ja) * 2007-10-24 2009-05-14 Crossfield Bio Inc 新規なラクトバチルス属微生物および哺乳動物用乳酸菌製剤
EP2110028A1 (fr) * 2008-04-15 2009-10-21 Nestec S.A. Expression de BDNF hippocampique et de Bifidobacterium longum
JP5511112B2 (ja) * 2011-06-14 2014-06-04 有限会社バイオメディカルリサーチグループ 菌体破砕物及びその配合物

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EP0555618A2 (fr) * 1992-02-10 1993-08-18 Renata Maria Anna Cavaliere Vesely Compositions diététiques ou pharmaceutiques comprenant des bactéries lactiques lyophilisées, leurs préparation et usage
JPH1192390A (ja) * 1997-09-17 1999-04-06 Natl Fedelation Of Agricult Coop Assoc ストレス状態改善剤
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EP0555618A2 (fr) * 1992-02-10 1993-08-18 Renata Maria Anna Cavaliere Vesely Compositions diététiques ou pharmaceutiques comprenant des bactéries lactiques lyophilisées, leurs préparation et usage
JPH1192390A (ja) * 1997-09-17 1999-04-06 Natl Fedelation Of Agricult Coop Assoc ストレス状態改善剤
EP1281752A1 (fr) * 2000-05-02 2003-02-05 Biofermin Pharmaceutical Co., Ltd. Cellules microbiennes sechees par pulverisation

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Title
HOSONO AKIRA ET AL.: "Comparison of the immunopotentiating activity with structural characteristics among water-soluble polysaccharides isolated from the genus bifidobacterium", BIOSCIENCE AND MICROFLORA, vol. 17, no. 2, 1998, pages 97 - 104, XP002982562 *
KOHO HIROKI ET AL.: "Kosoku stress fukaji no bifidobacterium kintai seibun toyo ni yoru chokan men'ekikei eno eikyo", NIHON NOGYO KAGAKUKAI KOEN YOSHISHU, vol. 2003, March 2003 (2003-03-01), pages 74, XP002982563 *
NAKANISHI YUSUKE ET AL.: "Bifidobacterium kintai seibun no mouse keiko toyo ni yoru chokan men'eki oto no kaiseki", THE JAPANESE SOCIETY OF NUTRITION AND FOOD SCIENCE SOKAI KOEN YOSHISHU, vol. 56, 2002, pages 351, XP002982561 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10028981B2 (en) 2009-08-25 2018-07-24 Nestec S.A. Bifidobacterium longum and functional GI disorders
US11452745B2 (en) 2009-08-25 2022-09-27 Societe Des Produits Nestle S.A. Bifidobacterium longum and functional GI disorders
US11957720B2 (en) 2009-08-25 2024-04-16 Societe Des Produits Nestle S.A. Bifidobacterium longum and functional GI disorders

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