CN115216423B - 一种动物双歧杆菌sf及其在医药和食品中的应用 - Google Patents
一种动物双歧杆菌sf及其在医药和食品中的应用 Download PDFInfo
- Publication number
- CN115216423B CN115216423B CN202210681663.9A CN202210681663A CN115216423B CN 115216423 B CN115216423 B CN 115216423B CN 202210681663 A CN202210681663 A CN 202210681663A CN 115216423 B CN115216423 B CN 115216423B
- Authority
- CN
- China
- Prior art keywords
- bifidobacterium animalis
- subsp
- lactis
- bifidobacteriumanimalis
- biologic
- 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.)
- Active
Links
- 241001134770 Bifidobacterium animalis Species 0.000 title claims abstract description 60
- 229940118852 bifidobacterium animalis Drugs 0.000 title claims abstract description 60
- 239000003814 drug Substances 0.000 title claims abstract description 14
- 235000013305 food Nutrition 0.000 title claims abstract description 14
- 229940079593 drug Drugs 0.000 title claims abstract description 9
- 230000000694 effects Effects 0.000 claims abstract description 28
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 24
- 230000000968 intestinal effect Effects 0.000 claims abstract description 19
- 206010012735 Diarrhoea Diseases 0.000 claims abstract description 18
- 230000006378 damage Effects 0.000 claims abstract description 8
- 230000001737 promoting effect Effects 0.000 claims abstract description 7
- 206010061218 Inflammation Diseases 0.000 claims abstract description 5
- 230000004054 inflammatory process Effects 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 4
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 claims description 16
- 238000002360 preparation method Methods 0.000 claims description 10
- 239000006041 probiotic Substances 0.000 claims description 10
- 235000018291 probiotics Nutrition 0.000 claims description 10
- 229960004768 irinotecan Drugs 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 206010009944 Colon cancer Diseases 0.000 claims description 7
- 238000009472 formulation Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 230000000529 probiotic effect Effects 0.000 claims description 5
- 239000004480 active ingredient Substances 0.000 claims description 4
- 235000013361 beverage Nutrition 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 230000036541 health Effects 0.000 claims description 3
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 2
- 235000013365 dairy product Nutrition 0.000 claims description 2
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 235000013311 vegetables Nutrition 0.000 claims description 2
- UYNVMODNBIQBMV-UHFFFAOYSA-N 4-[1-hydroxy-2-[4-(phenylmethyl)-1-piperidinyl]propyl]phenol Chemical compound C1CC(CC=2C=CC=CC=2)CCN1C(C)C(O)C1=CC=C(O)C=C1 UYNVMODNBIQBMV-UHFFFAOYSA-N 0.000 claims 1
- 229960003998 ifenprodil Drugs 0.000 claims 1
- 230000005764 inhibitory process Effects 0.000 abstract description 10
- 210000005027 intestinal barrier Anatomy 0.000 abstract description 4
- 230000007358 intestinal barrier function Effects 0.000 abstract description 3
- 230000004614 tumor growth Effects 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 23
- 241000186000 Bifidobacterium Species 0.000 description 13
- 241001465754 Metazoa Species 0.000 description 13
- 229940127093 camptothecin Drugs 0.000 description 13
- 229920002444 Exopolysaccharide Polymers 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 239000000047 product Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 210000001072 colon Anatomy 0.000 description 7
- 150000004676 glycans Chemical class 0.000 description 7
- 229920001282 polysaccharide Polymers 0.000 description 7
- 239000005017 polysaccharide Substances 0.000 description 7
- 238000010186 staining Methods 0.000 description 7
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 description 6
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 208000029742 colonic neoplasm Diseases 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 210000000952 spleen Anatomy 0.000 description 6
- 230000006907 apoptotic process Effects 0.000 description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 description 5
- 210000003734 kidney Anatomy 0.000 description 5
- 210000004185 liver Anatomy 0.000 description 5
- 239000002207 metabolite Substances 0.000 description 5
- 108090000695 Cytokines Proteins 0.000 description 4
- 102000004127 Cytokines Human genes 0.000 description 4
- 229920002307 Dextran Polymers 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108010087230 Sincalide Proteins 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- INAAIJLSXJJHOZ-UHFFFAOYSA-N pibenzimol Chemical compound C1CN(C)CCN1C1=CC=C(N=C(N2)C=3C=C4NC(=NC4=CC=3)C=3C=CC(O)=CC=3)C2=C1 INAAIJLSXJJHOZ-UHFFFAOYSA-N 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000000770 proinflammatory effect Effects 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102000004106 Claudin-3 Human genes 0.000 description 2
- 108090000599 Claudin-3 Proteins 0.000 description 2
- 206010010774 Constipation Diseases 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 102000003940 Occludin Human genes 0.000 description 2
- 108090000304 Occludin Proteins 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000001640 apoptogenic effect Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 230000001771 impaired effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 208000026775 severe diarrhea Diseases 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 230000005760 tumorsuppression Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000186018 Bifidobacterium adolescentis Species 0.000 description 1
- 241000186012 Bifidobacterium breve Species 0.000 description 1
- 241000186011 Bifidobacterium catenulatum Species 0.000 description 1
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 1
- 241001134772 Bifidobacterium pseudocatenulatum Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- 102000002029 Claudin Human genes 0.000 description 1
- 108050009302 Claudin Proteins 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 238000010867 Hoechst staining Methods 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940004120 bifidobacterium infantis Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 230000010428 chromatin condensation Effects 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 230000008951 colonic inflammation Effects 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000013872 defecation Effects 0.000 description 1
- 230000003544 deproteinization Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000002175 goblet cell Anatomy 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003871 intestinal function Effects 0.000 description 1
- 208000037817 intestinal injury Diseases 0.000 description 1
- 230000003903 intestinal lesions Effects 0.000 description 1
- 230000004673 intestinal mucosal barrier function Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000002417 nutraceutical Substances 0.000 description 1
- 235000021436 nutraceutical agent Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Images
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- 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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
-
- 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
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- 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
- A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
- A23L21/10—Marmalades; Jams; Jellies; Other similar fruit or vegetable compositions; Simulated fruit products
-
- 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
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- 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
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- A23V2400/515—Animalis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Food Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Nutrition Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种动物双歧杆菌SF,其生物保藏编号为CCTCC NO:M2021048。所述动物双歧杆菌SF可用于制备缓解伊利替康造成的副作用并促进其抑瘤作用的药物或保健品或食品,其可通过促进伊利替康抑制肿瘤增长的能力,并缓解伊利替康导致的腹泻,肠屏障破坏与肠道炎症来缓解伊利替康的副作用。
Description
技术领域
本发明属微生物领域,具体涉及一种动物双歧杆菌SF及在医药和食品中的其应用。
背景技术
伊利替康(CPT-11)是植物生物碱喜树碱的半合成水溶性衍生物,已被用于治疗多种类型的实体瘤。但在临床应用中常引起不良反应,如迟发性腹泻、呕吐、中性粒细胞减少等,限制了其临床应用。CPT-11在治疗过程中会产生肠道毒性,导致肠黏膜屏障受损,进而导致肠道功能下降,这些影响可能导致多达87%的患者出现延迟腹泻,其中30-40%的患者出现严重腹泻。近年来关于 CPT-11副作用机制研究以及预防和治疗,已成为国内外生物学、医药学研究的热点之一。
益生菌被定义为有活力的微生物,其中足够数量的益生菌以活跃状态到达肠道,从而对健康产生积极影响。益生菌可在肠道定殖,补充有益菌群,维持肠道菌群平衡,缓解疾病。有研究表明益生菌可通过调节肠道菌群缓解便秘、结肠炎等疾病。
双歧杆菌是最重要的益生菌之一,其对人体健康具有生物屏障、营养作用、抗肿瘤作用、免疫增强作用、改善胃肠道功能、抗衰老等多种重要的生理功能,广泛用于食品和医药行业。双歧杆菌有32个亚型,如双歧杆菌、长双歧杆菌、婴儿双歧杆菌、短双歧杆菌、青春双歧杆菌、梭形双歧杆菌、链状双歧杆菌、假链状双歧杆菌和齿状双歧杆菌等。众多研究已经证明双歧杆菌能抑制人体有害细菌的生长,抵抗病原菌的感染,合成人体需要的维生素,促进人体对矿物质的吸收,产生醋酸、丙酸、丁酸和乳酸等有机酸刺激肠道蠕动,促进排便,防止便秘以及抑制肠道腐败作用、净化肠道环境、分解致癌物质、刺激人体免疫系统,从而提高抗病能力等方面有着重要作用。但即属于同一个属,不同种的双歧杆菌,生物活性或者生物功能也各有不同。
发明内容
本发明的目的在于提供一种新的动物双歧杆菌SF,以及其在医药和食品中的应用。
本发明的第一个方面,是提供了一种能促进伊利替康抑瘤作用和缓解伊利替康副作用动物双歧杆菌SF。
一种动物双歧杆菌(Bifidobacterium animalis subsp.lactis)SF,其保藏号为CCTCC NO:M2021048。
本发明的第二个方面,是提供了上述动物双歧杆菌 (Bifidobacterium animalissubsp.lactis)SF的应用。
所述动物双歧杆菌(Bifidobacterium animalis subsp.lactis)SF或其代谢产物、或其提取液在制备预防和治疗结直肠癌的制剂中的应用。
所述动物双歧杆菌(Bifidobacterium animalis subsp.lactis)SF或其代谢产物、或其提取液在制备预防和治疗肠道炎症、肠道损伤和/或腹泻的制剂中的应用。
所述动物双歧杆菌(Bifidobacterium animalis subsp.lactis)SF或其代谢产物、或其提取液在制备缓解伊利替康副作用的制剂中的应用。
所述动物双歧杆菌(Bifidobacterium animalis subsp.lactis)SF或其代谢产物、或其提取液在制备促进伊利替康抑制肿瘤的作用的制剂中的应用。
在其中一些实施例中,所述制剂为药物或保健品或食品或者菌剂。
在其中一个实施例中,所述食品为乳制品、蔬菜制品、饮料制品和其他发酵类制品,例如饮料、酸奶或果冻。
所述保健品为液体状保健品、颗粒状保健品、粉末状保健品、胶囊保健品或片状保健品;或所述药品为溶液剂、颗粒剂、散剂、胶囊剂或片剂
在其中一个实施例中,所述副作用包括由伊利替康导致的肠道损伤和/或腹泻。
本发明的第三个方面,是提供了一种益生菌菌剂,其包括活性成分以及药学上或者食品上可接受的辅料,所述活性成分包括上述动物双歧杆菌 (Bifidobacteriumanimalis subsp.lactis)SF或其代谢产物、或其提取液。
在其中一个实施例中,所述益生菌菌剂由动物双歧杆菌SF制备而成,并以新鲜活菌菌体形式存在。
本发明所述的动物双歧杆菌SF,2021年1月12日保存于武汉市中国典型培养物保藏中心(China Center for Type Culture Collection,简称CCTCC M,地址:湖北省武汉市武昌区八一路299号武汉大学校内,邮编:430072),保藏编号为CCTCC NO:M 2021048。
本发明的发明人筛选到一株新的动物双歧杆菌SF(CCTCC NO:M 2021048),经过体外实验证明,本发明所述的动物双歧杆菌SF不但能抑制结肠癌细胞活性并促进其凋亡,而且,还能促进伊利替康的抑瘤作用,并且,经过动物实验证明,本发明动物双歧杆菌SF能有效缓解伊利替康造成的副作用,包括伊利替康导致的肠道损伤和/或腹泻,所述动物双歧杆菌SF(CCTCC M2021048)具有非常好的临床药用价值。
附图说明
图1是本发明所述动物双歧杆菌SF琼脂平板上生长的示意图。
图2是本发明所述动物双歧杆菌SF革兰氏染色100倍镜检的示意图。
图3是实施例1中所述细胞实验结果,动物双歧杆菌SF的胞外多糖可抑制结肠癌细胞活性并促进其凋亡;其中A为CCK-8法检测HCT-8细胞活性结果, B为CCK-8法检测Caco2细胞活性结果,C为Hoechst 33258染色HCT-8细胞的200倍镜检结果。
图4是实施例2中采用动物实验证明动物双歧杆菌SF促进伊利替康的抑瘤作用;其中A为反映8天中小鼠肿瘤体积变化的折线图,B为抑瘤率结果,C为小鼠肿瘤直拍图。
图5是实施例3中采用动物实验建模说明动物双歧杆菌SF影响了小鼠脾脏指数,对肝脏和肾脏指数没有影响,同时也减轻了小鼠腹泻;其中A为脾脏指数, B为肝脏指数,C为肾脏指数,D为腹泻评分。
图6是实施例4中所述动物实验结果,采用免疫荧光技术检测小鼠肿瘤组织中 CD4+与CD8+细胞的含量,说明动物双歧杆菌SF增加了小鼠肿瘤组织中CD4+与CD8+免疫细胞的含量。
图7是实例5中所述FITC-葡聚糖实验、实时荧光定量PCR与组化染色结果图,说明动物双歧杆菌SF缓解了伊利替康的肠道毒性,其中A为FITC浓度,B 为紧密连接蛋白的mRNA水平,C为结肠组织中促炎细胞因子的表达水平,D 为结肠组织中抗炎因子的表达水平,E为结肠组织的H&E染色,F为结肠组织的PAS染色。
具体实施方式
下面对本发明的实施例进行详细说明。
本发明提供了一种可缓解伊利替康副作用并促进其抑瘤作用的动物双歧杆菌SF,其可用于制备缓解伊利替康副作用并促进其抑瘤作用药物或保健品或食品。
以下实施例中所述的动物双歧杆菌SF,2021年1月12日保存于武汉市中国典型培养物保藏中心,保藏编号为CCTCC NO:M 2021048。
本发明所述的动物双歧杆菌SF的16s rDNA序列为:
gctccgctccatcgcatggtggggtgggaaatgcttttgcggcatgggatggggtcgcgtcctatcagcttgttggcgg ggtgatggcccaccaaggcgttgacgggtagccggcctgagagggtgaccggccacattgggactgagatacggcc cagactcctacgggaggcagcagtggggaatattgcacaatgggcgcaagcctgatgcagcgacgccgcgtgcggg atggaggccttcgggttgtaaaccgcttttgttcaagggcaaggcacggtttcggccgtgttgagtggattgttcgaataa gcaccggctaactacgtgccagcagccgcggtaatacgtagggtgcgagcgttatccggatttattgggcgtaaaggg ctcgtaggcggttcgtcgcgtccggtgtgaaagtccatcgcctaacggtggatctgcgccgggtacgggcgggctgga gtgcggtaggggagactggaattcccggtgtaacggtggaatgtgtagatatcgggaagaacaccaatggcgaaggc aggtctctgggccgtcactgacgctgaggagcgaaagcgtggggagcgaacaggattagataccctggtagtccacg ccgtaaacggtggatgctggatgtggggccctttccacgggtcccgtgtcggagccaacgcgttaagcatcccgcctg gggagtacggccgcaaggctaaaactcaaagaaattgacgggggcccgcacaagcggcggagcatgcggattaatt cgatgcaacgcgaagaaccttacctgggcttgacatgtgccggatcgccgtggagacacggtttcccttcggggcggtt cacaggtggtgcatggtcgtcgtcagctcgtgtcgtgagatgttggttaagtcccgcaacgagcgcaaccctcgccgca tgtgcagcggtgatgcggactcatgtggacgcgggtcactcgaggaggtgggatgacgtcagaatcatcatgccctac gttcagcctcacgcatgctacaatggcgtacacgcggtgcaacggtacctggggcggatccgcctgaaaaccggttct ctcagttccggataaagggtgttctaccttagacggctcccccacaagggtcgggccaccggcttcgggtgctacccac tttcatgacttgacgggcggtgtgtacaaggcccgggaacgcattcaccgcggcgttgctgatccgcgattactagcga ctccgccttcacgcagtcgagttgcagactgcgatccgaactgagaccggttttcagcgatccgccccacgtcaccgtg tcgcaccgcgttgtaccggccattgtagcatgcgtgaagccctggacgtaaggggcatgatgatctgacgtcatcccca ccttcctccgagttgaccccggcggtcccacatgagttcccggcatcacccgctggcaacatgcggcgagggttgcgc tcgttgcgggacttaacccaacatctcacgacacgagctgacgacgaccatgcaccacctgtgaaccggccccgaag ggaaaccgtgtctccacggcgatccggcacatgtcaagcccaggtaaggttcttcgcgttgcatcgaattaatccgcatg ctccgccgcttgtgcgggcccccgtcaatttctttgagttttagccttgcggccgtactccccaggcgggatgcttaacgc gttggctccgacacgggacccgtggaaagggccccacatccagcatccaccgtttacggcgtggactaccagggtatc taatcctgttcgctccccacgctttcgctcctcagcgtcagtgacggcccagagacctgccttcgccattggtgttcttccc gatatctacacattccaccgttacaccgggaattccagtctcccctaccgcactccagcccgcccgtacccggcgcaga tccaccgttaggcgatggactttcacaccggacgcgacgaaccgcctacgagccctttacgcccaataaatccggataa cgctcgcaccctacgtattaccgcggctgctggcacgtagttagccggtgcttattcgaacaatccactcaacacgggc gaaccgtgccttgcccttgaacaaaagcggtttacacccgaaggcctccatcccgcacgcgcgtcgctgcatcagcttg cgcccattgtgcatatcccactgctgctccgtagagtctggcgtatctcagtcccatgtgacgtcacctctcaagccggct aacgtcacgccttgctgcataccgccacagctgatgaaccggacctatccaagggcgccaagacatc(SEQ ID NO.1)。
本发明所述的动物双歧杆菌SF菌落小而凸圆、边缘完整,呈乳白色,表面光滑湿润有光泽,如图1所示。
本发明所述的动物双歧杆菌SF革兰氏染色100倍镜检,如图2所示动物双歧杆菌SF是革兰氏阳性杆菌,菌体长短不一,呈弯曲状、刮勺状。
实施例1动物双歧杆菌SF的胞外多糖可抑制结肠癌细胞活性并促进其凋亡
动物双歧杆菌SF的胞外多糖(EPS)的提取;(1)活化动物动物双歧杆菌SF,按接种量1%接至BS培养基,37℃厌氧培养24h;(2)8000r/min,离心10min,将上清转移至另一锥形瓶中;(3)发酵上清液加入3倍体积95%乙醇,4℃沉淀24h;(4)8000r/min,离心10min,弃上清,得沉淀,吹干后加入蒸馏水溶解;(5)Sevag法脱蛋白:加入1/3体积的Sevag液(氯仿:正丁醇=3:1),震荡30min,4000r/min,15min离心去沉淀;(6)重复步骤5,直到没有白色沉淀析出;(7)乙醇沉淀:加入3倍体积的95%乙醇,充分震荡至多糖如絮状沉淀析出,4000r/min,15min离心,收集沉淀;(8)多糖溶解:吹干沉淀,用5mL蒸馏水溶解,取2mL用14000Da透析袋透析72h;(9)吸取剩余的液体真空冷冻干燥得多糖。
小鼠结肠腺癌细胞株HCT-8和Caco2在含有10%胎牛血清、100U/mL 青霉素和100μg/mL链霉素的RPMI-1640培养基中于5%CO2的加湿培养箱中 37℃培养。
按照每孔约1×104HCT-8或Caco2细胞接种在96孔细胞培养板中。然后,在37℃、5%CO2的潮湿环境中孵育12小时后,在与上述相同的条件下,将动物双歧杆菌SF胞外多糖(EPS)与细胞在37℃下共孵育24小时和48小时。最后每孔加入10μL CCK-8试剂,37℃孵育1h后测定OD450。每个浓度占对照的百分比表示为细胞活力。
用Hoechst 33258染色后,在荧光显微镜下测定细胞凋亡的形态变化。细胞用不同浓度的EPS(0、100、200、400、800和1600μg/mL)处理48h,用 5mg/L Hoechst 33258在37℃染色30分钟,然后在标准激发的荧光显微镜下观察过滤器。凋亡细胞被定义为显示细胞核和细胞质收缩、染色质浓缩和凋亡小体的细胞。
实施例1结果如图3A与3B所示,不同浓度的动物双歧杆菌胞外多糖EPS 对HCT-8细胞(A)与Caco2(B)细胞的活性有一定的抑制作用,当处理时间为48h且EPS浓度达到1600μg/mL时,EPS对HCT-8细胞的抑制率高达41.2%,而浓度为800、1600μg/mL的EPS对Caco2细胞活性有明显的抑制作用,当EPS 浓度达到1600μg/mL,处理时间为48h时,抑制率达36.4%。参见图3C,对于 Hoechst染色实验,Control组的细胞核呈正常的蓝色,而加入了较高浓度EPS的处理组的部分细胞染色质固缩,呈致密浓染的状态,颜色发白,说明EPS可促进结肠癌细胞HCT-8凋亡。*P<0.05;**P<0.01。
实施例2:动物双歧杆菌SF促进了伊利替康的抑瘤作用
将-80℃冻存的植物乳杆菌动物双歧杆菌SF冻干粉用接种环挑取,于无菌 BS固体平板上划线活化,37℃厌氧培养24h复苏,经2次传代活化后转接无菌BS液体培养基,过夜培养。将活化2代后的动物双歧杆菌SF按2%接种于无菌BS液态培养基中,37℃厌氧培养24h后进行活菌计数。将培养液以 8000rpm离心3min后弃去上清液,根据活菌计数结果,用无菌1×PBS缓冲液将菌体制成1×109cfu/ml的菌液,每天制备一次菌液。
正常日粮预饲一周后,采用购自北京维通利华公司的Balb/c小鼠皮下注射肿瘤细胞构建荷瘤小鼠模型。根据肿瘤大小,将18只荷瘤小鼠分为三组,肿瘤(MD)组,给药(CPT)组和动物双歧杆菌辅助(SF)组。MD组与CPT 组小鼠灌胃无菌1×PBS,SF组小鼠灌胃动物双歧杆菌SF菌液(1×108cfu/mL) 0.1mL,连续灌胃8天。MD组每2天腹腔注射0.1mL生理盐水,CPT组与SF 组每2天腹腔注射0.1mL CPT-11溶液。实验总共持续8天,每天使用游标卡尺监测肿瘤大小,并通过以下公式计算:(宽度mm)2×(长度mm)/2。
实施例2结果如图4所示,在动物双歧杆菌SF作用下,SF组荷瘤小鼠的肿瘤体积增长速度明显低于单药CPT组,且抑瘤率显著提高。*P<0.05;***P <0.001。
实施例3:动物双歧杆菌SF影响了小鼠脾脏指数,对肝脏和肾脏指数没有影响,同时也减轻了小鼠腹泻
动物实验建模同实施例2。处死小鼠后对小鼠进行眼球采血,收集小鼠肝脏、肾脏和脾脏,计算脏器指数同时,通过腹泻评分标准,记录小鼠腹泻指数,0(正常),固体大便;1(轻度腹泻),略湿软便;2(中度腹泻),湿而未成形的大便,有中度的肛周染色;3(严重腹泻),水样大便,有严重的肛周染色。
结果如图5A-5C所示,与MD组相比,CPT组的脾脏指数显著升高,而肝脏指数与肾脏指数没有影响,说明CPT-11可能影响了小鼠的免疫系统,而 SF组的脾脏指数较之CPT组有更为显著的变化,这说明动物双歧杆菌SF的加入或许进一步加深了小鼠免疫系统的变化。同时,如图5D所示,CPT-11的摄入显著增加了MD组小鼠的腹泻评分,而动物双歧杆菌SF对腹泻有所缓解。
*P<0.05;**P<0.01;***P<0.001。
实施例4:动物双歧杆菌SF增加了小鼠肿瘤组织中CD4+与CD8+免疫细胞的含量
动物实验建模同实施例2。处死小鼠后收集小鼠肿瘤组织,采用免疫荧光技术检测小鼠CD4+与CD8+细胞的含量。
可以看出,CPT组的CD4+(红色荧光)和CD8+T(绿色荧光)细胞明显多于MD组。动物双歧杆菌SF干扰后,SF组CD4+(红色荧光)和CD8+T(绿色荧光)细胞多于CPT组。CD4+T细胞(辅助性T细胞)和CD8+T细胞(细胞毒性T细胞)是幼稚T细胞分化获得的两种类型,两者都是肿瘤免疫治疗过程中重要的T淋巴细胞。CD4+T细胞可协助其他淋巴细胞参与免疫反应,如识别抗原、释放调节性细胞因子、激活效应T细胞等,而CD8+T细胞可直接破坏异常组织细胞,或释放IL-2、IFN-γ等效应分子参与免疫反应。因此,动物双歧杆菌SF可以有效促进淋巴细胞的增殖和分化从而抑制肿瘤生长。**P< 0.01(图6)。
实施例5:动物双歧杆菌SF缓解了伊利替康的肠道毒性,包括肠道屏障受损、肠道炎症以及肠道病理损伤
动物实验建模同实施例2。处死小鼠后取实验小鼠的结肠组织,采用实时荧光定量PCR检测小鼠肠道紧密连接蛋白(zonula occludin-1、claudin-3和 occludin)和炎症因子(图7B-D),制作石蜡切片,进行HE染色和PAS染色(图7E-F),并通过FITC-葡聚糖实验(图7A)检测肠道通透性。如图7,与 MD组相比,CPT组FITC-葡聚糖浓度显著增加,而SF组FITC-葡聚糖浓度与 MD组相似。同时,与MD组相比,CPT-11治疗还降低了claudin-3、ZO-1和 occludin的mRNA水平,而SF组抑制了这种下降趋势。在CPT组中,结肠中促炎细胞因子(IL-6、TNF-α、IFN-γ和IL-1β)的mRNA表达显著上调,动物双歧杆菌SF干预显著下调这些促炎细胞因子的表达。此外,CPT-11处理还增加了TGF-β和IL-22的mRNA表达水平,同时降低了IL-10的表达水平。然而,动物双歧杆菌SF干预也显著下调TGF-β和IL-22的表达,并上调荷瘤小鼠中 IL-10的表达。结肠组织H&E和PAS染色显示,与MD组相比,CPT组肠道损伤明显,表现为炎症浸润和上皮增生,杯状细胞大量丢失。然而,SF组的炎症细胞浸润减少,表明动物双歧杆菌SF干预显着减轻了结肠炎症。*P<0.05; **P<0.01;***P<0.001;*is compared with CPT。
结果预示动物双歧杆菌SF可能在未来成为有效的缓解伊利替康造成的肠道毒性,包括肠道屏障受损、肠道炎症以及肠道病理损伤等副作用药物。
以上仅为本发明的具体实施例,并不以此限定本发明的保护范围;在不违反本发明构思的基础上所作的任何替换与改进,均属本发明的保护范围。
序列表
<110> 南昌大学
<120> 一种动物双歧杆菌SF及其在医药和食品中的应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2346
<212> DNA
<213> Bifidobacterium animalis(动物双歧杆菌)
<400> 1
gctccgctcc atcgcatggt ggggtgggaa atgcttttgc ggcatgggat ggggtcgcgt 60
cctatcagct tgttggcggg gtgatggccc accaaggcgt tgacgggtag ccggcctgag 120
agggtgaccg gccacattgg gactgagata cggcccagac tcctacggga ggcagcagtg 180
gggaatattg cacaatgggc gcaagcctga tgcagcgacg ccgcgtgcgg gatggaggcc 240
ttcgggttgt aaaccgcttt tgttcaaggg caaggcacgg tttcggccgt gttgagtgga 300
ttgttcgaat aagcaccggc taactacgtg ccagcagccg cggtaatacg tagggtgcga 360
gcgttatccg gatttattgg gcgtaaaggg ctcgtaggcg gttcgtcgcg tccggtgtga 420
aagtccatcg cctaacggtg gatctgcgcc gggtacgggc gggctggagt gcggtagggg 480
agactggaat tcccggtgta acggtggaat gtgtagatat cgggaagaac accaatggcg 540
aaggcaggtc tctgggccgt cactgacgct gaggagcgaa agcgtgggga gcgaacagga 600
ttagataccc tggtagtcca cgccgtaaac ggtggatgct ggatgtgggg ccctttccac 660
gggtcccgtg tcggagccaa cgcgttaagc atcccgcctg gggagtacgg ccgcaaggct 720
aaaactcaaa gaaattgacg ggggcccgca caagcggcgg agcatgcgga ttaattcgat 780
gcaacgcgaa gaaccttacc tgggcttgac atgtgccgga tcgccgtgga gacacggttt 840
cccttcgggg cggttcacag gtggtgcatg gtcgtcgtca gctcgtgtcg tgagatgttg 900
gttaagtccc gcaacgagcg caaccctcgc cgcatgtgca gcggtgatgc ggactcatgt 960
ggacgcgggt cactcgagga ggtgggatga cgtcagaatc atcatgccct acgttcagcc 1020
tcacgcatgc tacaatggcg tacacgcggt gcaacggtac ctggggcgga tccgcctgaa 1080
aaccggttct ctcagttccg gataaagggt gttctacctt agacggctcc cccacaaggg 1140
tcgggccacc ggcttcgggt gctacccact ttcatgactt gacgggcggt gtgtacaagg 1200
cccgggaacg cattcaccgc ggcgttgctg atccgcgatt actagcgact ccgccttcac 1260
gcagtcgagt tgcagactgc gatccgaact gagaccggtt ttcagcgatc cgccccacgt 1320
caccgtgtcg caccgcgttg taccggccat tgtagcatgc gtgaagccct ggacgtaagg 1380
ggcatgatga tctgacgtca tccccacctt cctccgagtt gaccccggcg gtcccacatg 1440
agttcccggc atcacccgct ggcaacatgc ggcgagggtt gcgctcgttg cgggacttaa 1500
cccaacatct cacgacacga gctgacgacg accatgcacc acctgtgaac cggccccgaa 1560
gggaaaccgt gtctccacgg cgatccggca catgtcaagc ccaggtaagg ttcttcgcgt 1620
tgcatcgaat taatccgcat gctccgccgc ttgtgcgggc ccccgtcaat ttctttgagt 1680
tttagccttg cggccgtact ccccaggcgg gatgcttaac gcgttggctc cgacacggga 1740
cccgtggaaa gggccccaca tccagcatcc accgtttacg gcgtggacta ccagggtatc 1800
taatcctgtt cgctccccac gctttcgctc ctcagcgtca gtgacggccc agagacctgc 1860
cttcgccatt ggtgttcttc ccgatatcta cacattccac cgttacaccg ggaattccag 1920
tctcccctac cgcactccag cccgcccgta cccggcgcag atccaccgtt aggcgatgga 1980
ctttcacacc ggacgcgacg aaccgcctac gagcccttta cgcccaataa atccggataa 2040
cgctcgcacc ctacgtatta ccgcggctgc tggcacgtag ttagccggtg cttattcgaa 2100
caatccactc aacacgggcg aaccgtgcct tgcccttgaa caaaagcggt ttacacccga 2160
aggcctccat cccgcacgcg cgtcgctgca tcagcttgcg cccattgtgc atatcccact 2220
gctgctccgt agagtctggc gtatctcagt cccatgtgac gtcacctctc aagccggcta 2280
acgtcacgcc ttgctgcata ccgccacagc tgatgaaccg gacctatcca agggcgccaa 2340
gacatc 2346
Claims (11)
1.一种动物双歧杆菌SF,其生物保藏编号为CCTCC NO:M 2021048。
2.权利要求1所述的动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF在制备治疗结直肠癌的制剂中的应用。
3.权利要求1所述的动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF在制备治疗伊利替康导致的肠道损伤和/或腹泻的制剂中的应用。
4.权利要求1所述的动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF在制备促进伊利替康抑制肿瘤的作用的制剂中的应用。
5.权利要求1所述的所述动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF在制备缓解伊利替康副作用的制剂中的应用;
所述副作用为由伊利替康导致的肠道炎症、肠道损伤和/或腹泻。
6.根据权利要2-5任一项所述的应用,其特征在于,所述制剂为药物或益生菌菌剂。
7.一种生物制剂,其特征在于,其包括活性成分以及药学上或者食品上可接受的辅料,所述活性成分包括权利要求1所述的动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF。
8.根据权利要求7所述的生物制剂,其特征在于,所述生物制剂为保健品或食品或益生菌菌剂。
9.根据权利要求7所述的生物制剂,其特征在于,所述生物制剂为药物。
10.根据权利要求8所述的生物制剂,其特征在于,所述食品为乳制品、蔬菜制品、饮料制品和发酵类制品。
11.根据权利要求8所述的生物制剂,其特征在于,所述益生菌菌剂中的所述动物双歧杆菌(Bifidobacteriumanimalis subsp. lactis)SF以新鲜活菌菌体形式存在。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210681663.9A CN115216423B (zh) | 2022-06-15 | 2022-06-15 | 一种动物双歧杆菌sf及其在医药和食品中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210681663.9A CN115216423B (zh) | 2022-06-15 | 2022-06-15 | 一种动物双歧杆菌sf及其在医药和食品中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115216423A CN115216423A (zh) | 2022-10-21 |
CN115216423B true CN115216423B (zh) | 2023-04-25 |
Family
ID=83607381
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210681663.9A Active CN115216423B (zh) | 2022-06-15 | 2022-06-15 | 一种动物双歧杆菌sf及其在医药和食品中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115216423B (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117327632B (zh) * | 2023-12-01 | 2024-02-13 | 四川厌氧生物科技有限责任公司 | 一种动物双歧杆菌及其应用 |
CN118272277B (zh) * | 2024-05-29 | 2024-09-20 | 善恩康生物科技(苏州)有限公司 | 一种能够缓解腹泻的动物双歧杆菌及其应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112618577A (zh) * | 2020-12-15 | 2021-04-09 | 深圳君拓生物科技有限公司 | 动物双歧杆菌在提高肿瘤免疫治疗应答中的作用 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108721337B (zh) * | 2017-04-17 | 2022-05-10 | 江苏德禧生物科技有限公司 | 一种预防肿瘤化疗肠道毒性的微生物菌剂 |
KR102563191B1 (ko) * | 2017-07-05 | 2023-08-02 | 에벨로 바이오사이언시즈, 인크. | 비피도박테리움 애니멀리스 ssp. 락티스를 이용한 암 치료 조성물 및 방법 |
-
2022
- 2022-06-15 CN CN202210681663.9A patent/CN115216423B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112618577A (zh) * | 2020-12-15 | 2021-04-09 | 深圳君拓生物科技有限公司 | 动物双歧杆菌在提高肿瘤免疫治疗应答中的作用 |
Also Published As
Publication number | Publication date |
---|---|
CN115216423A (zh) | 2022-10-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110496140B (zh) | 脆弱拟杆菌或阿克曼粘细菌在制备用于预防或治疗肿瘤的药物中的应用 | |
CN115216423B (zh) | 一种动物双歧杆菌sf及其在医药和食品中的应用 | |
Kato et al. | Effects of oral administration of Lactobacillus casei on antitumor responses induced by tumor resection in mice | |
CN110893195B (zh) | 一种具有缓解肠道炎症功能的副干酪乳杆菌et-22 | |
CN110101722A (zh) | 一种复合益生菌菌剂用于制备治疗溃疡性结肠炎产品的用途 | |
CN105434476A (zh) | 一种脆弱拟杆菌在预防和/或治疗炎症性肠病中的应用 | |
WO2017020784A1 (zh) | 一种脆弱拟杆菌及其应用 | |
CN106413724B (zh) | 与多糖聚合物粘合剂缀合的鼠李糖乳杆菌rht-3201及其用途 | |
CN110893194B (zh) | 乳双歧杆菌bl-99在抑制肠道炎症方面的新应用 | |
US20080003207A1 (en) | Treating inflammatory bowel disease with live bacteria | |
CN114259056A (zh) | 一种鼠李糖乳酪杆菌在制备用于预防和/或治疗溃疡性结肠炎的食品或药品中的应用 | |
CN114470003B (zh) | 脆弱拟杆菌或其两性离子荚膜多糖在制备防治消化系统肿瘤药物中的应用 | |
CN111011856A (zh) | 用于缓解胃病的组合物、其制备方法及用于缓解胃病的食品 | |
CN116121154B (zh) | 一种乳明串珠菌及其应用 | |
CN114681493B (zh) | 动物双歧杆菌乳亚种的应用 | |
CN115607577B (zh) | 一种具有缓解口腔炎症功效的益生菌剂及其制备方法和应用 | |
CN112746034A (zh) | 一种协同抑制幽门螺旋杆菌的鼠李糖乳杆菌lr863和鼠李糖乳杆菌lr519及其应用 | |
CN114558036A (zh) | 脆弱拟杆菌在改善和治疗腹泻中的应用 | |
CN114344325A (zh) | 脆弱拟杆菌及其两性离子荚膜多糖在制备用于防治生殖泌尿系统肿瘤的药物中的应用 | |
CN114107134A (zh) | 一株侧孢短芽孢杆菌及其应用 | |
CN116445356B (zh) | 一种调节肠道菌群及增强免疫力的动物双歧杆菌乳亚种ba67及其应用 | |
CN117264814A (zh) | 一种对消化道疾病具有预防治疗效果的鼠李糖乳酪杆菌 | |
CN116694534A (zh) | 一种长双歧杆菌sx-1326及其应用 | |
CN115517367B (zh) | 副干酪乳杆菌smn-lbk在制备促进肠道健康产品中的应用 | |
CN114796283B (zh) | 五谷虫提取物用于制备治疗消化性疾病药物中的应用及药物组合物 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |