CN112574904A - Bifidobacterium V9 for regulating immunity and application thereof - Google Patents
Bifidobacterium V9 for regulating immunity and application thereof Download PDFInfo
- Publication number
- CN112574904A CN112574904A CN202011084880.7A CN202011084880A CN112574904A CN 112574904 A CN112574904 A CN 112574904A CN 202011084880 A CN202011084880 A CN 202011084880A CN 112574904 A CN112574904 A CN 112574904A
- Authority
- CN
- China
- Prior art keywords
- bifidobacterium
- immunity
- effect
- increasing
- product
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000186000 Bifidobacterium Species 0.000 title claims abstract description 99
- 230000036039 immunity Effects 0.000 title claims abstract description 17
- 230000001105 regulatory effect Effects 0.000 title claims abstract description 8
- 230000002708 enhancing effect Effects 0.000 claims abstract description 9
- 210000004698 lymphocyte Anatomy 0.000 claims description 20
- 241000699670 Mus sp. Species 0.000 claims description 16
- 102000003814 Interleukin-10 Human genes 0.000 claims description 13
- 108090000174 Interleukin-10 Proteins 0.000 claims description 13
- 230000009466 transformation Effects 0.000 claims description 9
- 206010020751 Hypersensitivity Diseases 0.000 claims description 8
- 230000003393 splenic effect Effects 0.000 claims description 6
- 208000026935 allergic disease Diseases 0.000 claims description 5
- 230000007815 allergy Effects 0.000 claims description 5
- 210000003736 gastrointestinal content Anatomy 0.000 claims description 4
- 238000009629 microbiological culture Methods 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 230000035755 proliferation Effects 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 2
- 241001591005 Siga Species 0.000 claims 1
- 235000013361 beverage Nutrition 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 235000013305 food Nutrition 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 230000001939 inductive effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 48
- 239000006041 probiotic Substances 0.000 abstract description 8
- 235000018291 probiotics Nutrition 0.000 abstract description 8
- 108020000411 Toll-like receptor Proteins 0.000 description 15
- 102000002689 Toll-like receptor Human genes 0.000 description 15
- 241000699666 Mus <mouse, genus> Species 0.000 description 13
- 229940076144 interleukin-10 Drugs 0.000 description 12
- 210000000822 natural killer cell Anatomy 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 11
- 238000003304 gavage Methods 0.000 description 11
- 239000002609 medium Substances 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 230000037396 body weight Effects 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 102000006992 Interferon-alpha Human genes 0.000 description 7
- 108010047761 Interferon-alpha Proteins 0.000 description 7
- 230000003111 delayed effect Effects 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 238000003556 assay Methods 0.000 description 6
- 210000000952 spleen Anatomy 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 5
- 108090000695 Cytokines Proteins 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 102000003945 NF-kappa B Human genes 0.000 description 4
- 108010057466 NF-kappa B Proteins 0.000 description 4
- 210000003719 b-lymphocyte Anatomy 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 241001494479 Pecora Species 0.000 description 3
- 208000030961 allergic reaction Diseases 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 230000036737 immune function Effects 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000001976 improved effect Effects 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 206010011968 Decreased immune responsiveness Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 2
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 2
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 244000144993 groups of animals Species 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 210000000548 hind-foot Anatomy 0.000 description 2
- 230000028996 humoral immune response Effects 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 230000004957 immunoregulator effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 2
- 230000008719 thickening Effects 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- NCGICGYLBXGBGN-UHFFFAOYSA-N 3-morpholin-4-yl-1-oxa-3-azonia-2-azanidacyclopent-3-en-5-imine;hydrochloride Chemical group Cl.[N-]1OC(=N)C=[N+]1N1CCOCC1 NCGICGYLBXGBGN-UHFFFAOYSA-N 0.000 description 1
- 101710168515 Cell surface glycoprotein Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 229920002444 Exopolysaccharide Polymers 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 102100026720 Interferon beta Human genes 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000013462 Interleukin-12 Human genes 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 102000003816 Interleukin-13 Human genes 0.000 description 1
- 108090000176 Interleukin-13 Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 102000000743 Interleukin-5 Human genes 0.000 description 1
- 102000004890 Interleukin-8 Human genes 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000030852 Parasitic disease Diseases 0.000 description 1
- 108010013639 Peptidoglycan Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 101710099182 S-layer protein Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 210000000447 Th1 cell Anatomy 0.000 description 1
- 102000008228 Toll-like receptor 2 Human genes 0.000 description 1
- 108010060888 Toll-like receptor 2 Proteins 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000007124 immune defense Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 229940099472 immunoglobulin a Drugs 0.000 description 1
- 239000003547 immunosorbent Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000004719 natural immunity Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 230000036281 parasite infection Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 210000001986 peyer's patch Anatomy 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108010009004 proteose-peptone Proteins 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
Abstract
The invention discloses a bifidobacterium V9 for regulating immunity and application thereof. The bifidobacterium V9 has high viable count, and a lower amount of strains can play the role of enhancing immunity. Compared with commercial probiotics, the bifidobacterium V9 has high cost performance, is more substantial, and has obvious effect of improving immunity.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to a bifidobacterium V9 for regulating immunity and application thereof.
Background
The immune enhancing effects of probiotics have been widely recognized. In productive life, the most common probiotics are bifidobacteria, which are important physiological bacteria in human intestinal tract and have physiological effects incomparable with various other normal physiological bacteria: regulating intestinal flora, enhancing immunity, etc.
The bifidobacterium plays a role in enhancing the immune defense system of the body by promoting and activating infection protection immune response of the body. The bifidobacterium colonizes in the intestinal tract, is equivalent to natural immunity, can activate the macrophage activity of the organism, induces specific and nonspecific immune reaction, and improves the anti-infection capacity; meanwhile, the bifidobacterium adjusts the relationship among the flora through the interaction between the bifidobacterium and other bacteria and metabolites thereof, maintains and ensures the optimal advantage combination and the stability of various combinations of the flora, prevents the colonization and invasion of pathogenic bacteria, and antagonizes the growth of the pathogenic bacteria and harmful microorganisms and the adhesion of toxins thereof.
Immunoregulatory ability has received widespread attention in recent years as a standard for screening probiotics and an important function of probiotics. Since 90% of diseases are related to immunity, the level of immunity directly determines whether a person is healthy or not. The immune system plays an important role in resisting the invasion of external harmful substances, so that more and better substances for improving the immunoregulatory capacity of the body are sought, and the immune system plays an important role in the future of researching and developing new health-care products.
Disclosure of Invention
The invention provides a bifidobacterium V9 for regulating immunity, which can solve one or more of the problems in the prior art.
Peptidoglycan, teichoic acid, lipoteichoic acid, S-layer protein and other cell wall associated polysaccharides are present on the surface of bifidobacteria. In addition, bifidobacteria also secrete substances with immunomodulatory activity, such as exopolysaccharides and other secreted proteins, extracellularly.
The viable count of Bifidobacterium V9 of the present invention is not less than 5.8 × 1011The bifidobacterium V9 is preserved in China general microbiological culture collection center with the preservation number: CGMCC No.4473, preservation date: 12/14/2010, storage address: china general microbiological culture Collection center.
The application detects cytokines, Toll-like receptors (TLRs), nuclear factor kappa B (NF-kappa B) and secretory immunoglobulin (sIgA) at the mRNA level and the protein level, researches the immune regulation capacity and the action mechanism of bifidobacterium, and provides a basis for developing a microecological preparation.
The invention also provides application of the bifidobacterium V9 in enhancing the immunity of the organism. The application evaluates the influence of bifidobacterium V9 on the immune function of a healthy mouse after being fed with the bifidobacterium V9, and detects Toll-like receptors, NF-kappa B and sIgA factor expression by observing the weight, delayed allergic reaction, lymphocyte transformation experiment, natural killer cell (NK cell) activity and using an ELISA kit of the mouse. The results show that bifidobacterium V9 of the invention can induce lymphocyte proliferation, enhance delayed type allergy and the transformation of splenic lymphocytes of mice; the bifidobacterium V9 can improve the activity of NK cells of the tested animals; the bifidobacterium V9 can improve the transcription of interleukin-10 (IL-10) and interferon-alpha (IFN-alpha) cell factors, can also improve the expression of NF-kB, has up-regulation effect on TLR-2 and sIgA, and is beneficial to resisting the infection of intestinal pathogenic microorganisms. Therefore, the bifidobacterium V9 has a positive effect on improving the immunity of the organism.
Upon entry into the gut, bifidobacterium V9 can be taken up by M cells in the peyer's patch and presented to the Antigen Presenting Cells (APC) within it. Bifidobacterium V9 can induce lymphocyte to secrete cytokines such as IL-4, IL-5, IL-10, IL-13 and transforming growth factor-beta (TGF-beta), promote differentiation of helper T cell 2(Th2), activate B lymphocyte to produce immunoglobulin E (IgE), mainly mediate humoral immune response, and play a role in resisting parasite infection in body. The optimal dose of Bifidobacterium has opposite regulation effects, and can inhibit Th2 differentiation, shift to Th1 differentiation, promote B lymphocyte to produce sIgA, and inhibit IgE expression. The transformed Th1 cell secretes cytokines such as IL-2, IL-8, IFN-alpha, IFN-gamma, IFN-beta, TNF-alpha, IL-12 and granulocyte-macrophage colony stimulating factor (GM-CSF), can activate cytotoxic T lymphocyte, enhance phagocyte activity, mainly mediate cell immune response, and play an important role in inhibiting tumor and virus. The high dose of Bifidobacterium can produce clone anergy and cause T lymph to generate anergy.
The bifidobacterium of the invention can improve the immune function of the organism by two aspects: firstly, the innate immune response can be improved, macrophages are activated, and pathogenic bacteria are treated by producing active oxygen and lysosomes; the bifidobacteria also activate the adaptive immune system, activate B cells, secrete a range of antibodies, and participate in humoral immune responses. In cellular immunity, bifidobacteria can enhance the phagocytic function of macrophages and can also activate the activity of T cells and NK cells to stimulate the body to generate immune response. In humoral immunity, bifidobacteria can enhance the ability of B lymphocytes to secrete antibodies.
The bifidobacterium V9 has high viable count, and is supported by a large amount of experimental data and proved by clinical tests, and toxicological experiments and safety evaluation are carried out. And a lower amount of the strain can exert the immune enhancement function. Compared with commercial probiotics, the bifidobacterium V9 has high cost performance, is more substantial, and has obvious effect of improving immunity.
Drawings
FIG. 1 is a schematic diagram of the preparation of Bifidobacterium strain V9 in accordance with the present invention;
FIG. 2 is a graph showing the effect of Bifidobacterium V9 of the present invention on IL-10;
FIG. 3 is a graph showing the effect of Bifidobacterium V9 of the present invention on IFN-. alpha.;
FIG. 4 is a graph of the effect of Bifidobacterium V9 of the present invention on TLR-2;
FIG. 5 is a graph showing the effect of Bifidobacterium V9 of the present invention on NF- κ B;
FIG. 6 shows the effect of Bifidobacterium V9 of the present invention on sIgA.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
Fig. 1 schematically shows a schematic preparation of bifidobacterium V9 in accordance with an embodiment of the invention.
As shown in the figure, after the freeze-dried strain is activated for three generations, the freeze-dried strain is inoculated into an optimized culture medium for fermentation at 6% (v/v), and then the strain mud is obtained by centrifugal collection.
Preparing sterilized protective agent (skim milk powder), uniformly mixing the bacterial sludge and the protective agent according to the proportion (W/V) to obtain organic liquid, carrying out vacuum freeze drying to obtain freeze-dried coarse powder, crushing and sieving the coarse powder to obtain freeze-dried bifidobacterium V9 fine powder, subdividing bifidobacterium V9, inspecting, packaging, and warehousing for later use.
In the preparation method of the bifidobacterium V9, the optimized specific culture medium comprises 14 parts of lactose, 7 parts of beef extract, 14 parts of casein peptone, 6 parts of soybean peptone, 6 parts of yeast powder, 2 parts of disodium hydrogen phosphate, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate and 1 part of L-cysteine hydrochloride by weight.
Experimental samples: the living bacteria number of the bifidobacterium stems V9 of the invention can reach 5.8 x 1011/g。
Experimental animals: 48 SPF-grade Kunming female mice are selected and aged for 6-8 weeks (the weight is 18-22 g).
Dose and group: four groups of 12 animals each including a control group and low, medium and high dose groups were divided into 5 times (0.25 g/kg. BW), 10 times (0.5 g/kg. BW) and 30 times (1.5 g/kg. BW) of the recommended daily amount (0.05 g/kg. BW in terms of 30kg body weight of children).
Example 1
The feeding method comprises the following steps: 0.50 g, 1.00 g and 3.00g of bifidobacterium V9 sample are respectively weighed and prepared into 20mL of sample liquid by purified water, and the sample liquid is respectively used for intragastric administration of low, medium and high dose groups of animals. After animals were acclimated for three days in laboratory conditions, the following experiments each randomly divided 48 mice into four groups of 12 mice each, administered the test substance by gavage daily, the amount of the administered sample was adjusted according to the increase and decrease of body weight per week, and gavage was continued for four weeks. Sterile purified water was drenched to the control group.
Example 2
Delayed type allergic reaction test in mice (increase of plantar thickness)
On day 4 before the end of the experiment, the animals were immunized, 2% (v/v) sheep red blood cells were intraperitoneally injected with 0.2mL of sensitized animals, and after 5 days, the thickness of the plantar region of the left hind foot was measured, followed by subcutaneous injection of 20% (v/v) sheep red blood cells (20. mu.L/mouse) at that position, and the thickness of the plantar region of the left hind foot was measured three times 24 hours after the injection to obtain an average value.
Example 3
ConA-induced splenic lymphocyte transformation assay (MTT method) in mice
Lymphocyte proliferation reaction: the lymphocyte suspension was added to a 24-well plate in two wells, each well containing 1mL of the lymphocyte suspension, 75. mu.L of ConA solution (equivalent to 7.5. mu.g/mL) in one well, and 5% CO in the other well as a light2And culturing at 37 ℃ for 72 h. 4 hours before the end of the culture, 0.7mL of the supernatant was aspirated from each well, and 0.7mL of serum-free RPMI 1640 medium was added thereto together with 50. mu.L/well of MTT (5mg/mL), and the culture was continued for 4 hours. After the culture was completed, 1mL of isopropyl alcohol was added to each well, and the mixture was blown and beaten to dissolve the purple crystals, and the purple crystals were divided into 96-well plates as 3-well replicates, and the optical density value was measured at a wavelength of 570 nm.
Example 4
NK cell Activity assay (LDH method)
And (3) detecting the activity of NK cells: taking the concentration as 4 x 105YAC-1 target cells and effector cells of 100. mu.L each per mL (50: 1 ratio of effect to target) were added to a U-shaped 96-well plate; target cell natural release holes are filled with 100 mu L of target cells and culture solution respectively, and target cell maximum release holes are filled with 100 mu L of target cells and 100 mu L of 1% NP40 respectively; all the above-mentioned materials are equipped with three composite holes, at 37 deg.C and 5% CO2After 4 hours of incubation in an incubator, the 96-well plates were centrifuged at 1500rpm for 5 minutes, 100. mu.L of the supernatant was aspirated into each well and placed in a flat-bottomed 96-well plate, 100. mu.L of LDH matrix solution was added thereto, and the reaction was carried out for 3 minutes with 30. mu.L of HCl added at 1mol/L per well, and the optical density was measured at 492nm on an enzyme-linked immunosorbent.
Example 5
Cytokine, Toll-like receptor (TLR-2), NF-. kappa.B and sIgA assays
(1) And (4) separating serum. Four groups of animals in example 1 were removed and blood was collected. Treating blood in the centrifuge tube in 37 deg.C water bath for 10min, standing at 4 deg.C for 15min, and centrifuging at 3000rpm/min for 20 min. Carefully pipette the clear supernatant into a small clean centrifuge tube and store at-40 ℃ for use.
(2) And (4) detecting the cell factors. After thawing the frozen serum at room temperature, the following cytokines were detected according to the instructions of the ELISA test kit: IL-10, IFN-alpha.
(3) Toll-like receptor (TLR-2) and NF-. kappa.B assays: the method comprises the steps of adopting a cell freezing and thawing method, namely repeatedly freezing and thawing the separated mouse spleen lymphocytes for more than three times at room temperature and-40 ℃, centrifugally collecting supernate, and detecting each component by using an ELISA kit.
(4) And (3) sIgA detection: collecting and treating the intestinal contents of the mice according to the operation method of the kit, and detecting the contents in the contents according to the instruction of the ELISA detection kit.
Discussion of results
1. Animal experiments
1.1 Effect of Bifidobacterium V9 on mouse body weight
Effect of bifidobacterium V9 on mouse body weight: the delayed type allergic reaction group is shown in Table 1, and the lymphocyte transformation and NK activity assay group is shown in Table 2. The difference between the body weight of each dose group at each stage and the body weight of the control group at the corresponding stage has no significance, which indicates that the bifidobacterium V9 has no obvious influence on the body weight growth of the mice.
Table 1 effect of bifidobacterium V9 on mouse body weight (batch I, n ═ 12)
Table 2 effect of bifidobacterium V9 on mouse body weight (batch II, n ═ 12)
1.2 Effect of Bifidobacterium V9 on mouse spleen weight
Compared with the control group, the difference of the organ indexes of each dose group has no significant significance (see table 3). It was shown that bifidobacterium V9 had no significant effect on the spleen weight of the mice.
Table 3 effect of bifidobacterium V9 on mouse spleen weight (n ═ 12)
1.3 Effect of Bifidobacterium V9 on mouse Immunity
1.3.1 Effect of Bifidobacterium V9 on delayed allergy in mice sheep Red blood cell-induced delayed allergy test in mice (plantar thickening method), the plantar thickening values of the test subjects were significantly different from those of the control group (see Table 4).
Table 4 effect of probiotic electuary on delayed allergy in mice (n ═ 12)
1.3.2 Effect of Bifidobacterium V9 on ConA-induced splenic lymphocyte transformation in mice
For the ConA-induced splenic lymphocyte transformation reaction of mice, the difference of the optical density of the test object group is higher than that of the control group, and the difference has significant significance (see table 5). The result of the spleen lymphocyte transformation test of the bifidobacterium V9 is positive, and the tested sample has the function of enhancing the cellular immune function.
TABLE 5 Effect of Bifidobacterium on mouse splenic lymphocyte transformation response (n ═ 12)
1.3.3 Effect of Bifidobacterium V9 on NK cell Activity of test animals
The NK cell activity of the medium and high dose groups is higher than that of the control group, and the difference has significance (see table 6). The result of the test on the NK cell activity of the tested animal is positive by the bifidobacterium V9, and the tested sample has the function of enhancing the NK cell activity.
NK cell activity conversion value: x is Sin-1(p)1/2
Table 6 effect of probiotic infusion on NK cell activity in mice (n ═ 11)
2. Effect of Bifidobacterium V9 on IL-10
The effect of Bifidobacterium V9 on IL-10 levels in serum is shown in FIG. 2. After the gavage, the content of IL-10 can be improved by the bifidobacterium V9 in all the dose groups, after 20 days of the gavage, the content of IL-10 is obviously improved by the bifidobacterium V9 in low, medium and high doses (p is less than 0.05), and the differences of the content of IL-10 from 700pg/mL to 798pg/mL, 899pg/mL and 956pg/mL in the control group are respectively obvious (p is less than 0.05). The results show that the bifidobacterium V9 can increase the expression level of IL-10 in the body.
3. Effect of Bifidobacterium V9 on IFN-alpha
The effect of bifidobacterium V9 on IFN- α content in serum is shown in figure 3. After the gavage, the content of IL-10 can be increased by the bifidobacterium V9 in all dose groups, after 20 days of the gavage, the content of IFN-alpha (p is less than 0.05) is remarkably increased by the bifidobacterium V9 in low, medium and high doses, and the differences are remarkable (p is less than 0.05) compared with a control group respectively from 625pg/mL to 789pg/mL, 887pg/mL and 923pg/mL in the control group. The results show that the bifidobacterium V9 can increase the expression level of IFN-alpha of the organism and has obvious influence.
4. Effect of Bifidobacterium V9 on TLR-2
The effect of bifidobacterium V9 on Toll-like receptor 2(TLR-2) expression by mouse spleen lymphocytes is shown in figure 4. Bifidobacterium V9(14.7ng/mL) in the high dose group at day 10 of gavage significantly increased TLR-2 expression in lymphocytes (p < 0.05). Bifidobacterium V9 in the medium dose group also increased TLR-2 expression, but the results were not significant; on day 20 of gavage, bifidobacterium V9(15.1ng/mL) in the high dose group significantly increased TLR-2 expression in lymphocytes (p < 0.05). Bifidobacterium V9 in the medium dose group also increased TLR-2 expression, but the results were not significant;
5. effect of Bifidobacterium V9 on NF- κ B
The effect of Bifidobacterium V9 on NF-. kappa.B expression levels is shown in FIG. 5. On the 10 th day of gavage, bifidobacterium V9(10892pg/mL) in the high dose group significantly increased NF- κ B expression (p < 0.05) compared to the control group. Bifidobacterium V9 in the medium dose group also increased NF- κ B expression, but the results were not significant; on day 20 of gavage, bifidobacterium V9(14564pg/mL) in the high dose group significantly increased NF- κ B expression (p < 0.05), while bifidobacterium V9 in the medium dose group had no significant effect.
6. Effect of Bifidobacterium V9 on sIgA
The results of the intestinal mucosal secretory immunoglobulin A (sIgA) assay are shown in FIG. 6. On the 10 th day of gavage, all doses of bifidobacterium V9 increased the content of sIgA in the intestinal contents compared to the control group (50 ng/mL). The results show that bifidobacterium V9 can improve the concentration of sIgA in intestinal contents, particularly the concentration of sIgA reaches a peak value after 10 days of intragastric administration. In addition, bifidobacterium V9 in the high dose group did not produce more sIgA than in the medium dose group.
What has been described above are merely some embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made without departing from the inventive concept thereof, and these changes and modifications can be made without departing from the spirit and scope of the invention.
Claims (9)
1. A Bifidobacterium V9 for regulating immunity, wherein the viable count of Bifidobacterium V9 is not less than 5.8 × 1011The bifidobacterium V9 is preserved in China general microbiological culture Collection center with the following preservation numbers: CGMCC No. 4473.
2. A Bifidobacterium V9 formulation for modulating immunity comprising Bifidobacterium V9 as claimed in claim 1.
3. Use of bifidobacterium V9 in a product for increasing the expression level of IL-10 in a body according to claim 1.
4. Use of bifidobacterium V9 in a product for increasing the expression level of IFN- α in the body according to claim 1.
5. Use of bifidobacterium V9 in accordance with claim 1 in products for inducing lymphocyte proliferation, enhancing delayed-type allergy and enhancing transformation of splenic lymphocytes in mice.
6. Use of bifidobacterium V9 in a product for increasing the expression of NF- κ B according to claim 1.
7. Use of bifidobacterium V9 in a product for increasing the expression of TLR-2 in lymphocytes, as claimed in claim 1.
8. Use of bifidobacterium V9 in a product for increasing the content of sIgA in intestinal contents according to claim 1.
9. Use of the bifidobacterium V9 in foods, pharmaceuticals and beverages for regulating immunity according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011084880.7A CN112574904A (en) | 2020-10-12 | 2020-10-12 | Bifidobacterium V9 for regulating immunity and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011084880.7A CN112574904A (en) | 2020-10-12 | 2020-10-12 | Bifidobacterium V9 for regulating immunity and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112574904A true CN112574904A (en) | 2021-03-30 |
Family
ID=75120223
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011084880.7A Pending CN112574904A (en) | 2020-10-12 | 2020-10-12 | Bifidobacterium V9 for regulating immunity and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112574904A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102326678A (en) * | 2011-06-16 | 2012-01-25 | 高杰 | Penta-composite probiotic powder preparation and preparation method thereof |
| US20120207713A1 (en) * | 2007-03-28 | 2012-08-16 | Alimentary Health Limited | Probiotic bifidobacterium strains |
| CN106212674A (en) * | 2016-08-05 | 2016-12-14 | 内蒙古普泽生物制品有限责任公司 | A kind of Yoghourt fermentation mix bacterium agent |
-
2020
- 2020-10-12 CN CN202011084880.7A patent/CN112574904A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120207713A1 (en) * | 2007-03-28 | 2012-08-16 | Alimentary Health Limited | Probiotic bifidobacterium strains |
| CN102326678A (en) * | 2011-06-16 | 2012-01-25 | 高杰 | Penta-composite probiotic powder preparation and preparation method thereof |
| CN106212674A (en) * | 2016-08-05 | 2016-12-14 | 内蒙古普泽生物制品有限责任公司 | A kind of Yoghourt fermentation mix bacterium agent |
Non-Patent Citations (1)
| Title |
|---|
| 翁梁等, 中国轻工业出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114032193B (en) | Lactobacillus paracasei 207-27 and application thereof | |
| CN110964657B (en) | Bifidobacterium lactis BL-99 capable of improving immunity and application thereof | |
| CN104894021A (en) | Lactobacillus paracasei strain and application thereof | |
| CN101328468B (en) | Antiallergic lactic acid bacteria | |
| JP6337262B2 (en) | Salt-resistant lactic acid bacteria, culture method of salt-resistant lactic acid bacteria, and immunostimulant | |
| CN113142301B (en) | Dairy product capable of improving immunity of organism and preparation method thereof | |
| Li et al. | Suppressive effects of oral administration of heat-killed Lactobacillus acidophilus on T helper-17 immune responses in a bovine β-lactoglobulin-sensitized mice model | |
| CN113005067B (en) | Multifunctional composite probiotic preparation and preparation method thereof | |
| WO2023168808A1 (en) | Lactobacillus paracasei capable of regulating symptoms of intestinal immune disorders and use thereof | |
| JP4112021B2 (en) | Immunostimulant using lactic acid bacteria | |
| CN110484459B (en) | Lactobacillus plantarum and application thereof | |
| CN114703108A (en) | Lactobacillus mucilaginosus strain and application thereof in improvement of facial redness and type I rosacea | |
| JP2617758B2 (en) | Antibody production enhancer | |
| CN114381395A (en) | Lactobacillus plantarum ZJUFN1 and application thereof | |
| CN111685255B (en) | Probiotic solid beverage for enhancing immune function and preparation method thereof | |
| CN111254087B (en) | Lactobacillus helveticus with high adhesion performance and function of enhancing immunity and application thereof | |
| KR102525173B1 (en) | Bacillus bacteria, interleukin-22 production inducer, skin barrier function enhancer | |
| Shukla et al. | Protective potential of L. acidophilus in murine giardiasis | |
| Kim et al. | Immune-enhancing effects of Lactobacillus plantarum 200655 isolated from Korean Kimchi in a cyclophosphamide-induced immunocompromised mouse model | |
| CN112574904A (en) | Bifidobacterium V9 for regulating immunity and application thereof | |
| CN102028224B (en) | Antiallergic lactobacillus | |
| Eze et al. | Effects of Lactobacillus spp. isolated from the sap of palm tree Elaeis guineensis (palm wine) on cellular and innate immunity | |
| CN113005066B (en) | Compound bifidobacterium preparation for resisting allergy, increasing immunity, reducing blood sugar and fat and losing weight and preparation method thereof | |
| CN111728030B (en) | Sucrose-free yogurt with immunity improving function and long shelf life at normal temperature and preparation method thereof | |
| CN111743158B (en) | Probiotic tablet with function of enhancing immunity and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210330 |