CN116870037A - Application of bifidobacterium adolescentis ATCC15703 or soluble polysaccharide component thereof in preparation of anti-aging pharmaceutical food or preparation targeting intestinal tract - Google Patents
Application of bifidobacterium adolescentis ATCC15703 or soluble polysaccharide component thereof in preparation of anti-aging pharmaceutical food or preparation targeting intestinal tract Download PDFInfo
- Publication number
- CN116870037A CN116870037A CN202310130338.8A CN202310130338A CN116870037A CN 116870037 A CN116870037 A CN 116870037A CN 202310130338 A CN202310130338 A CN 202310130338A CN 116870037 A CN116870037 A CN 116870037A
- Authority
- CN
- China
- Prior art keywords
- intestinal
- bifidobacterium adolescentis
- aging
- soluble polysaccharide
- atcc15703
- 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.)
- Granted
Links
- 241001486432 Bifidobacterium adolescentis ATCC 15703 Species 0.000 title claims abstract description 64
- 150000004676 glycans Chemical class 0.000 title claims abstract description 30
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 30
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 30
- 235000013305 food Nutrition 0.000 title claims description 19
- 238000002360 preparation method Methods 0.000 title claims description 18
- 230000003712 anti-aging effect Effects 0.000 title claims description 12
- 210000001035 gastrointestinal tract Anatomy 0.000 title claims description 12
- 230000008685 targeting Effects 0.000 title claims description 8
- 210000001072 colon Anatomy 0.000 claims abstract description 38
- 230000032683 aging Effects 0.000 claims abstract description 32
- 230000000968 intestinal effect Effects 0.000 claims abstract description 30
- 210000004027 cell Anatomy 0.000 claims abstract description 19
- 210000004966 intestinal stem cell Anatomy 0.000 claims abstract description 15
- 230000019491 signal transduction Effects 0.000 claims abstract description 8
- 210000002220 organoid Anatomy 0.000 claims description 49
- 230000014509 gene expression Effects 0.000 claims description 22
- 108090000623 proteins and genes Proteins 0.000 claims description 17
- 102000013814 Wnt Human genes 0.000 claims description 15
- 108050003627 Wnt Proteins 0.000 claims description 15
- 230000012010 growth Effects 0.000 claims description 13
- 210000000130 stem cell Anatomy 0.000 claims description 10
- 239000003814 drug Substances 0.000 claims description 9
- 210000001519 tissue Anatomy 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 7
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 230000004156 Wnt signaling pathway Effects 0.000 claims description 6
- 230000035784 germination Effects 0.000 claims description 5
- 230000028327 secretion Effects 0.000 claims description 4
- 206010061218 Inflammation Diseases 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims description 3
- 235000013402 health food Nutrition 0.000 claims description 3
- 230000004054 inflammatory process Effects 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 2
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 2
- 230000003213 activating effect Effects 0.000 claims description 2
- 235000013361 beverage Nutrition 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 235000013365 dairy product Nutrition 0.000 claims description 2
- 235000013325 dietary fiber Nutrition 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 10
- 239000006041 probiotic Substances 0.000 abstract description 6
- 235000018291 probiotics Nutrition 0.000 abstract description 6
- 230000007358 intestinal barrier function Effects 0.000 abstract description 4
- 238000012423 maintenance Methods 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 230000018109 developmental process Effects 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 3
- 230000006870 function Effects 0.000 abstract description 2
- 210000005027 intestinal barrier Anatomy 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 11
- 101150017554 LGR5 gene Proteins 0.000 description 10
- 238000010186 staining Methods 0.000 description 7
- 230000009758 senescence Effects 0.000 description 6
- 241000186018 Bifidobacterium adolescentis Species 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 230000009471 action Effects 0.000 description 5
- 230000001737 promoting effect Effects 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 3
- 241000520272 Pantoea Species 0.000 description 3
- 108010013639 Peptidoglycan Proteins 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 206010010774 Constipation Diseases 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 241000736262 Microbiota Species 0.000 description 2
- 102000014736 Notch Human genes 0.000 description 2
- 108010070047 Notch Receptors Proteins 0.000 description 2
- 208000025174 PANDAS Diseases 0.000 description 2
- 208000021155 Paediatric autoimmune neuropsychiatric disorders associated with streptococcal infection Diseases 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000112 colonic effect Effects 0.000 description 2
- 210000002175 goblet cell Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000007166 healthy aging Effects 0.000 description 2
- 238000003125 immunofluorescent labeling Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000004609 intestinal homeostasis Effects 0.000 description 2
- 238000011813 knockout mouse model Methods 0.000 description 2
- 108010082117 matrigel Proteins 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 206010013554 Diverticulum Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- 101150058357 Muc2 gene Proteins 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- 102000003940 Occludin Human genes 0.000 description 1
- 108090000304 Occludin Proteins 0.000 description 1
- 240000000220 Panda oleosa Species 0.000 description 1
- 235000016496 Panda oleosa Nutrition 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 101150103187 Reg4 gene Proteins 0.000 description 1
- 102000040739 Secretory proteins Human genes 0.000 description 1
- 108091058545 Secretory proteins Proteins 0.000 description 1
- 108010017842 Telomerase Proteins 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 201000009840 acute diarrhea Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 208000019902 chronic diarrheal disease Diseases 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 230000008045 co-localization Effects 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000021045 dietary change Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000007366 host health Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 230000004207 intestinal integrity Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000012134 supernatant fraction Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 235000019722 synbiotics Nutrition 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- 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
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D8/00—Methods for preparing or baking dough
- A21D8/02—Methods for preparing dough; Treating dough prior to baking
- A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
-
- 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
- A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- 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
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- Animal Husbandry (AREA)
- Toxicology (AREA)
- Physiology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Agronomy & Crop Science (AREA)
- Botany (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of probiotics, and particularly relates to application of bifidobacterium adolescentis ATCC15703 or polysaccharide components thereof. The bifidobacterium adolescentis ATCC15703 or the soluble polysaccharide component thereof provided by the invention can activate a wnt signal pathway to promote the self-renewal potential of intestinal stem cells by improving the number and the functions of panid cells in colon, so that the intestinal aging state taking an intestinal barrier as a center can be remarkably improved; the bifidobacterium adolescentis ATCC15703 or the soluble polysaccharide component thereof has obvious effect of improving intestinal aging, has definite target intestinal aging resisting mechanism, can be used as probiotics for clinical application in the field of health maintenance to relieve the aging process of organisms, and provides a new intervention means for preventing or resisting aging progress and reducing the occurrence and development of aging-related diseases.
Description
Technical Field
The invention relates to the technical field of probiotics, in particular to application of bifidobacterium adolescentis ATCC15703 or soluble polysaccharide components thereof in preparation of anti-aging pharmaceutical foods or preparations targeting intestinal tracts.
Background
The active health ability of the old is improved, the concept of healthy aging and active aging is built, and the healthy gateway is moved forward to the old with low age. In face of the challenges presented by aging, the prevention of aging has become more important and urgent to help the elderly to achieve healthy aging. Therefore, it is urgent to explore a high-efficiency and low-cost control method.
Intestinal epithelium is one of the fastest-renewing tissues in the human body, which protects the host initially from various harmful environmental factors and pathogens. Intestinal homeostasis has profound effects on multiple systems. Self-renewal of intestinal stem cells is an important process to maintain intestinal integrity. Previous studies have shown that aging of the intestinal tract is accompanied by dysfunction and failure of ISCs, which further increases the incidence of age-related intestinal disorders such as chronic constipation, diverticulosis of the colon, malnutrition and colorectal cancer. Thus, the search for a new strategy to promote ISCs regeneration might provide a clear prospect for regeneration of aged intestines.
There is increasing evidence that the intestinal microbiota undergoes tremendous changes over the lifetime. Unbalanced changes in microbial composition and related metabolites have been shown to be age-related. This age-related microbial dysbiosis further triggers a cascade of inflammatory events, leading to intestinal leakage and affecting the overall health of the host. A recent study showed that fecal microbiota transplantation can attenuate the accelerated aging phenotype and extend the longevity of pre-matured mice. This provides an opportunity for slowing or even reversing age-related changes and diseases through microbiological interventions such as probiotics, synbiotics, dietary changes to microbiota, etc.
Bifidobacterium adolescentis is a completely anaerobic gram-positive bacterium in the normal intestinal tract of an adult, can autonomously reproduce, maintain the balance of intestinal flora, regulate the intestinal homeostasis of the human body, and is proved to be capable of relieving acute and chronic diarrhea and constipation, improving inflammatory bowel disease and delaying aging. Given the outstanding manifestations of bifidobacteria adolescentis in improving overall ageing, their protective effect on the ageing colon is still unknown.
Disclosure of Invention
The first object of the present invention is to provide the use of bifidobacterium adolescentis ATCC15703 or of the soluble polysaccharide fraction (SPS) thereof for the preparation of an anti-ageing pharmaceutical food or preparation targeting the intestinal tract.
The invention can also adopt or combine the following technical proposal when adopting the technical proposal:
as a preferred technical scheme of the invention: the intestine-targeted anti-aging pharmaceutical food or formulation prevents and improves the intestinal aging performance by at least one of the following:
1) Improving the intestinal aging performance, including mucosal thickness, crypt depth, colon stem cell density, and inflammation level;
2) Significantly improving the sprouting rate and growth status of the intestinal derived colon organoids;
3) Increasing the number of panid cells in an intestinal stem cell supporting environment;
4) Activating wnt signaling pathway in intestinal stem cells.
As a preferred technical scheme of the invention: the equivalent viable count of bifidobacterium adolescentis ATCC15703 or soluble polysaccharide component (SPS) in the intestine-targeted anti-aging pharmaceutical food or preparation is not less than 10 9 CFU/g or 10 9 CFU/mL。
As a preferred technical scheme of the invention: the anti-aging pharmaceutical food targeting the intestinal tract is a medicine, a health food or a food.
As a preferred technical scheme of the invention: the medicine comprises a medicine carrier and/or pharmaceutically acceptable auxiliary materials
As a preferred technical scheme of the invention: the food includes special medical food, solid beverage, dietary fiber, dairy product, bean product, cake or animal feed.
As a preferable technical scheme of the invention, the anti-aging medicines, the health-care foods and the food preparations are liquid preparations, capsule preparations or compound tablets.
The second object of the present invention is to provide the use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for the preparation of wnt family secretion protein expression promoters or wnt signal pathway expression promoters or panid cell expression promoters.
A third object of the present invention is to provide the use of Bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing the germination rate and growth status of an intestinal-derived colon organoid.
A fourth object of the present invention is to provide the use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing the level of wnt signalling pathways in colon stem cells.
A fifth object of the present invention is to provide the use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing the number of panid cells in intestinal tissue and derived colon organoids.
The bifidobacterium adolescentis ATCC15703 or the soluble polysaccharide component thereof provided by the invention can activate a wnt signal pathway to promote the self-renewal potential of intestinal stem cells by improving the number and the functions of panid cells in colon, so that the intestinal aging state taking an intestinal barrier as a center can be remarkably improved; the bifidobacterium adolescentis ATCC15703 or the soluble polysaccharide component thereof has obvious effect of improving intestinal aging, has definite target intestinal aging resisting mechanism, can be used as probiotics for clinical application in the field of health maintenance to relieve the aging process of organisms, and provides a new intervention means for preventing or resisting aging progress and reducing the occurrence and development of aging-related diseases. Compared with the prior art, the invention has at least the following advantages or beneficial effects:
(1) Bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof improves intestinal age-improving performance from many aspects: including mucosal thickness, crypt depth, colon stem cell density, and inflammation level; at present, no related literature and patent report exists, and the traditional Chinese medicine composition can be used as a beneficial supplement of the current anti-aging probiotic treatment means targeting the intestinal tract.
(2) Bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide component thereof is capable of significantly improving the germination rate and growth status of intestinal derived colon organoids in vitro and in vivo.
(3) Bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof is able to increase wnt signalling pathway levels in colon stem cells.
(4) Bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof is capable of significantly increasing the number of panid cells in intestinal tissue and derived organoids in vitro and in vivo.
(5) The bifidobacterium adolescentis ATCC15703 or the soluble polysaccharide component thereof takes colon Pantoea cells as a downstream important target spot, promotes the secretion of wnt protein families, further activates colon stem cells, and provides a new clue and a new idea for exploring intestinal tract aging in the future.
Drawings
FIG. 1 is a graphical representation of the effect of bifidobacterium adolescentis ATCC15703 on restoring intestinal stem cells to alleviate the ageing intestinal phenotype in example 1; in the figure, A is H & E staining in the intestinal tract after intervention of bifidobacterium adolescentis ATCC15703, muc2 staining, PAS staining, B.a bacterial FISH probe staining, lgr5 staining and staining of barrier protein (ZO-1, occludin); b is the transcript level of the colonic mucosa height and senescence-associated genes (P21, P53) and the stem cell gene Lgr 5.
FIG. 2 is a graphical representation of the effect of bifidobacterium adolescentis ATCC15703 on promoting proliferation of colon organoids derived from aging mice in example 2. In the figure, A is a schematic drawing of colon organoid extraction; B. c is the action and quantitative statistics of bifidobacterium adolescentis ATCC15703 on colon organoids of the G3 aging model mice respectively; D. e is the action and quantitative statistics of bifidobacterium adolescentis ATCC15703 on colon organoids of a natural aging model mouse respectively; f is the transcript level of the dry gene Lgr5 of the two organoid models.
FIG. 3 is a graphical representation of the effect of bifidobacterium adolescentis ATCC15703 in example 3 on increasing wnt signaling in colon organoids. FIG. A is a transcript level of the wnt signaling pathway-related gene after intervention of bifidobacterium adolescentis ATCC15703 in the colon organoid; B. c, D are the expression levels of important target genes of wnt signaling pathway after intervention of bifidobacterium adolescentis ATCC15703 on colon organoids, respectively.
FIG. 4 is a graphical representation of the effects of Pantoea adolescentis cells in example 4 on the maintenance of dryness of bifidobacterium adolescentis ATCC 15703. In the figure, A is the expression level of wnt secretion protein family after intervention of bifidobacterium adolescentis ATCC15703 in colon organoid; b is the expression level of a panduran cell-related gene after intervention of bifidobacterium adolescentis ATCC15703 in colon organoids; c is the fluorescent co-localization of bifidobacterium adolescentis ATCC15703 in the intervention of colon organoid stem cells and Pantoea inodorsum cells; d is the expression correlation of the pannocyte-like gene in the elderly population.
FIG. 5 is a schematic representation of the soluble polysaccharide fraction (SPS) effect fraction of bifidobacterium adolescentis ATCC15703 in example 5. In the figure, A is a schematic diagram of an extraction flow of main coarse components of bifidobacterium adolescentis; B. c, D the action and quantitative statistics of the broken supernatant and sediment of the bifidobacterium adolescentis on the colon organoids of the mice; E. f, G the action of the broken soluble polysaccharide component, lipid component and peptidoglycan component of Bifidobacterium adolescentis on colon organoids of mice and quantitative statistics are carried out respectively.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way.
Example 1: bifidobacterium adolescentis ATCC15703 relieves the ageing intestinal phenotype by recovering intestinal stem cells
The generation 0 Terc heterozygote mice are used for continuously breeding the generation 0 Terc knockout mice, which are used for constructing a third generation telomerase knockout mouse (G3 premature senility mice) model and littermates thereof, and the experiments are divided into 3 groups. Three groups were perfused with PBS, PBS and Bifidobacterium adolescentis ATCC15703, at a dose of 1X 10 9 CFU/mouse, the amount of lavage was 200. Mu.L, and the lavage was performed once every other day. After the modeling is completed, the mice are dissected, the intestinal length is counted, and the mice are processed through H&E staining shows intestinal epithelial morphology, shows barrier function related indexes such as goblet cell number by PAS staining and Muc2 immunofluorescence staining, shows stem cell number by Lgr5 immunofluorescence staining, and determines the spatial localization of bifidobacterium adolescentis ATCC15703 by a FISH probe.
The results shown in FIG. 1A, in which bifidobacterium adolescentis ATCC15703 was able to up-regulate the crypt height, increase PAS+ goblet cells and up-regulate the levels of Occlutin and ZO-1 in G3 presenility model mice, and the results shown in FIG. 1B, in which bifidobacterium adolescentis ATCC15703 down-regulates the expression of senescence-associated genes p21, p53 by about 50%, suggest that bifidobacterium adolescentis ATCC15703 can improve intestinal barrier function under senescence conditions. At the same time, bifidobacterium adolescentis ATCC15703 likewise increases Lgr5 + ISCs numbers and up-regulation of Lgr5 gene expression levels to 200% suggest that improvement of intestinal barrier function is mediated by intestinal stem cells.
Example 2: bifidobacterium adolescentis ATCC15703 for improving growth of various colon organoid senescence models
To further determine the effect of promoting bifidobacterium adolescentis ATCC 15703. As shown in fig. 2 a, colon organoids of G3 old mice and 12 month old natural aging mice were extracted, respectively: a1.5 cm sample of the colon tissue of the mouse was taken and the intestinal contents were flushed with sterile PBS. The intestinal segments were cut, and the tissue was cut into small pieces and then placed in 10mM EDTA solution and digested with shaking at 4℃for 30min at 10-20 rpm. The samples were centrifuged and resuspended in 10mL pre-chilled DMEM/F-12 medium and counted, mixed with matrigel containing complete organoid growth medium and inoculated in 24 well plates. After the matrigel is solidified, 500 mu L of complete culture medium is added, and the mixture is placed at 37 ℃ and 5% CO 2 Culturing in an incubator. After the third day organoids were grown stable, the intestinal organoids were co-cultured with bifidobacterium adolescentis ATCC15703 (moi=100:1), followed by observation of organoid growth indicators (diameter and bud number) on the fifth and seventh days. At the end of the seventh day of growth, organoid RNA was extracted and assayed for Lgr5 transcript levels.
The results are shown in fig. 2 a and B: bifidobacterium adolescentis ATCC15703 has obvious proliferation promoting effect on colon crypt organoids derived from two aging model mice, and organoid germination rate and diameter are increased by about 20-50%. The results are shown in fig. 2C: the increased transcript level of the Lgr5 gene in both derived organoids was about 20%, indicating that bifidobacterium adolescentis ATCC15703 has an improving effect against the colonic organoid senescence model in a variety of senescence models.
Example 3: bifidobacterium adolescentis ATCC15703 increases organoid wnt signalling pathway levels
To explore the mechanism of promotion by bifidobacterium adolescentis ATCC15703, the colon organoids of mice were co-cultured with bifidobacterium adolescentis ATCC15703 (moi=100:1) as described previously, and RNA was extracted on day seven. The expression of the genes related to the signal paths such as wnt, notch and the like is detected by qPCR, immunofluorescence and western blotting.
As shown in FIGS. 3A and B, bifidobacterium adolescentis ATCC15703 increased the transcriptional expression level of the wnt signaling pathway target genes CyclinD1, c-Myc to 150% -200% by detecting the wnt and notch pathway gene expression heat maps. As shown in FIG. 3C, immunofluorescence experiments showed that Bifidobacterium adolescentis ATCC15703 increases the proportion of active-beta-catenin nuclear-entering fluorescence in organoids. As shown in fig. 3D, bifidobacterium adolescentis ATCC15703 can significantly increase the expression level of key proteins of wnt signaling pathway to 2-4 fold at the protein level.
Example 4: the maintenance of organoid dryness by bifidobacterium adolescentis ATCC15703 is mediated by panid cells
Panda-like cells are considered as effective supporting cells in the microenvironment of intestinal stem cells, and can promote the growth and development of the supporting intestinal stem cells through the action mode similar to panda cells. Expression levels of wnt secretory protein family genes in colon organoids after intervention by bifidobacterium adolescentis ATCC15703 were analyzed by qPCR. The expression of the pannocyte-related genes (Lyz, reg 4) was then further examined in tissue samples and organoid samples.
As shown in FIGS. 4A and B, bifidobacterium adolescentis ATCC15703 can increase the transcription level and protein expression level of Lyz, reg4 in organoids and bodies. As shown in fig. 4C, immunofluorescence data showed that bifidobacterium adolescentis ATCC15703 can increase the relative intensity of Lyz and lgr5 fluorescence of organoids and intestinal tissues in vivo, suggesting that it can increase panid cell expression in vivo as well as in vitro. In a clinical human colon sample, the pandura-like related genes Lyz, reg4 and age are also found to have negative correlation, the Lyz and age correlation coefficient is as high as r= -0.7014, and the Reg4 and age correlation coefficient is as high as r= -0.3929.
Example 5: acquisition and effectiveness verification of soluble polysaccharide component of bifidobacterium adolescentis ATCC15703
In order to explore the role of the thallus components in the dryness promoting effect, the thallus components are separated and purified. Inoculating and culturing Bifidobacterium adolescentis ATCC157037 to logarithmic phase, centrifuging to remove supernatant, heat inactivating (95 deg.C, 15-30 min), ultrasonic (50-60 kH) and trichloroacetic acid (4-10%) to break bacterial cells, releasing content, co-culturing supernatant and precipitate with intestinal organoids respectively, and observing organoid growth index (diameter and bud number) on fifth and seventh days. At the end of the seventh day of growth, organoid RNA was extracted and assayed for Lgr5 transcript levels. Further, extracting and purifying lipid components from the crushed mixture by using an organic solvent, deproteinizing by using a plurality of proteases, and obtaining a peptidoglycan shell component after precipitation and cleaning; and simultaneously, obtaining the crude extract of the soluble polysaccharide by adopting a water extraction and alcohol precipitation method (volume ratio is 1:3). Finally, the bifidobacterium adolescentis ATCC15703 soluble polysaccharide component is obtained through dialysis and freeze-drying. These three components were co-cultured with the intestinal organoids separately, and organoid growth indicators (diameter and bud number) were then observed on the fifth and seventh days. At the end of the seventh day of growth, organoid RNA was extracted and assayed for Lgr5 transcript levels.
As shown in fig. 5 a, organoid co-culture experiments were performed after the end of the procedure for crude component extraction of bifidobacterium adolescentis ATCC 15703. As shown in FIGS. 5, B, C and D, the supernatant of Bifidobacterium adolescentis ATCC15703 in the disrupted fraction and the crude fraction of the sediment, the supernatant fraction has a promoting effect on mouse-derived crypt organoids, and the results show that it increases the germination rate and diameter by about 150% -200%, and increases the level of Lgr5, ascl2 dry gene expression by about 2-3 times. As shown in FIGS. 5, E, G and F, the lipid, peptidoglycan and soluble polysaccharide components of Bifidobacterium adolescentis ATCC15703 promote mouse-derived crypt organoids, which result in increased sprouting rates and diameters of about 150% and increased Lgr5, ascl2 dry gene expression levels by about 2-fold.
The above detailed description is intended to illustrate the present invention by way of example only and not to limit the invention to the particular embodiments disclosed, but to limit the invention to the precise embodiments disclosed, and any modifications, equivalents, improvements, etc. that fall within the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. Use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for the preparation of an anti-ageing pharmaceutical food or preparation targeting the intestinal tract.
2. The use according to claim 1, wherein the intestine-targeted anti-aging pharmaceutical food or formulation prevents and improves the intestinal aging state by at least one of the following:
1) Improving the intestinal aging performance, including mucosal thickness, crypt depth, colon stem cell density, and inflammation level;
2) Significantly improving the sprouting rate and growth status of the intestinal derived colon organoids;
3) Increasing the number of panid cells in an intestinal stem cell supporting environment;
4) Activating wnt signaling pathway in intestinal stem cells.
3. The use according to claim 1, characterized in that: the anti-aging pharmaceutical food targeting the intestinal tract is a medicine, a health food or a food.
4. A use according to claim 3, characterized in that: the medicine comprises a medicine carrier and/or pharmaceutically acceptable auxiliary materials.
5. A use according to claim 3, characterized in that: the food includes special medical food, solid beverage, dietary fiber, dairy product, bean product, cake or animal feed.
6. A use according to claim 3, characterized in that: the medicine or health food or the formulation of the food is liquid preparation, capsule preparation or compound tablet.
7. Use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for the preparation of a wnt family secretion protein expression promoter or a wnt signal pathway expression promoter or a panid cell expression promoter.
8. Use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing the germination rate and growth status of intestinal derived colon organoids.
9. Use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing wnt signalling pathway levels in colon stem cells.
10. Use of bifidobacterium adolescentis ATCC15703 or a soluble polysaccharide fraction thereof for increasing the number of panid cells in intestinal tissue and derived colon organoids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310130338.8A CN116870037B (en) | 2023-02-03 | 2023-02-03 | Application of soluble polysaccharide component of bifidobacterium adolescentis ATCC15703 in preparation of anti-aging medicine targeting intestinal tract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310130338.8A CN116870037B (en) | 2023-02-03 | 2023-02-03 | Application of soluble polysaccharide component of bifidobacterium adolescentis ATCC15703 in preparation of anti-aging medicine targeting intestinal tract |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116870037A true CN116870037A (en) | 2023-10-13 |
| CN116870037B CN116870037B (en) | 2024-06-28 |
Family
ID=88268564
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310130338.8A Active CN116870037B (en) | 2023-02-03 | 2023-02-03 | Application of soluble polysaccharide component of bifidobacterium adolescentis ATCC15703 in preparation of anti-aging medicine targeting intestinal tract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116870037B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190112674A1 (en) * | 2017-10-17 | 2019-04-18 | Infinitus (China) Company Ltd. | Bifidobacterium adolescentis and use thereof |
| CN110368406A (en) * | 2019-08-20 | 2019-10-25 | 江南大学 | Bifidobacterium adolescentis CCFM1062 is preparing the application in functional microbial inoculum, food and/or drug |
| CN112708581A (en) * | 2021-01-26 | 2021-04-27 | 青岛东海药业有限公司 | Bifidobacterium adolescentis DH162 and fermentation product, application and microecological preparation thereof |
| CN113005060A (en) * | 2021-03-15 | 2021-06-22 | 江南大学 | Application of bifidobacterium adolescentis CCFM1173 in preparation of functional microbial inoculum, food and/or medicament |
| CN113521111A (en) * | 2021-03-16 | 2021-10-22 | 浙江大学 | Application of bifidobacterium adolescentis in delaying senescence |
-
2023
- 2023-02-03 CN CN202310130338.8A patent/CN116870037B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190112674A1 (en) * | 2017-10-17 | 2019-04-18 | Infinitus (China) Company Ltd. | Bifidobacterium adolescentis and use thereof |
| CN110368406A (en) * | 2019-08-20 | 2019-10-25 | 江南大学 | Bifidobacterium adolescentis CCFM1062 is preparing the application in functional microbial inoculum, food and/or drug |
| CN112708581A (en) * | 2021-01-26 | 2021-04-27 | 青岛东海药业有限公司 | Bifidobacterium adolescentis DH162 and fermentation product, application and microecological preparation thereof |
| CN113005060A (en) * | 2021-03-15 | 2021-06-22 | 江南大学 | Application of bifidobacterium adolescentis CCFM1173 in preparation of functional microbial inoculum, food and/or medicament |
| CN113521111A (en) * | 2021-03-16 | 2021-10-22 | 浙江大学 | Application of bifidobacterium adolescentis in delaying senescence |
Non-Patent Citations (3)
| Title |
|---|
| FAN等: "B. adolescentis ameliorates chronic colitis by regulating Treg/Th2 response and gut microbiota remodeling", GUT MICROBES, vol. 13, no. 1, pages 2 * |
| 顾瑞霞;: "乳酸菌与免疫调节作用", 乳业科学与技术, no. 06, 1 November 2008 (2008-11-01), pages 251 - 256 * |
| 顾瑞霞;: "乳酸菌与免疫调节作用", 乳业科学与技术, no. 06, pages 251 - 256 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116870037B (en) | 2024-06-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115300531B (en) | Lactobacillus paracasei JY062 composition and preparation method and application thereof | |
| CN113897302B (en) | Bifidobacterium capable of relieving colitis and application thereof | |
| CN114574390A (en) | Bifidobacterium longum subspecies of infant for relieving colitis and application | |
| CN114107088B (en) | Lactobacillus reuteri LRSY523 and application thereof | |
| WO2023134212A1 (en) | New use of bacteroides fragilis and/or zwitterionic capsular polysaccharide of bacteroides fragilis | |
| CN115992059B (en) | Lactobacillus johnsonii for producing feruloyl esterase and application thereof in relieving ulcerative colitis | |
| CN116731929A (en) | Lactobacillus mucilaginosus ZS40 and application thereof | |
| CN117064920A (en) | Application of Akkermansia muciniphila in preparation of products for preventing, treating and/or assisting in treating tumors | |
| CN116870037B (en) | Application of soluble polysaccharide component of bifidobacterium adolescentis ATCC15703 in preparation of anti-aging medicine targeting intestinal tract | |
| CN115612652B (en) | Lactobacillus plantarum AS21 and application thereof in preventing ulcerative colitis | |
| CN115418332A (en) | Lactobacillus plantarum capable of preventing and improving chemical liver injury | |
| CN115011518A (en) | Lactobacillus mixture with effect of relieving colorectal cancer related to colitis and application thereof | |
| CN116019839A (en) | Application of enterococcus lactis JDM1 in preparation of medicines for preventing or treating inflammatory bowel disease | |
| CN112646743B (en) | Lactobacillus reuteri CCFM1134 for preventing and relieving ulcerative colitis and application thereof | |
| CN115806901A (en) | Lactobacillus brevis and application thereof in resisting cervical cancer | |
| Luo et al. | Protective effect of Enterococcus faecium against ethanol-induced gastric injury via extracellular vesicles | |
| CN115025128A (en) | Application of nocardia rubra cell wall skeleton in treatment of cervical lesion | |
| CN114854623A (en) | Lactobacillus delbrueckii subspecies bulgaricus, microbial inoculum containing same and application | |
| CN113773983A (en) | Bifidobacterium longum subspecies longum strain and application thereof | |
| CN117965401B (en) | Lactobacillus rhamnosus AFY01 and application of product thereof in inflammatory colon cancer | |
| CN115554340B (en) | Peony seed meal extract for treating liver cancer by synergic chemotherapy drugs and preparation method and application thereof | |
| CN113679741B (en) | Application of human amniotic epithelial stem cells in preparation of medicines for treating cisplatin-induced acute kidney injury | |
| CN115109718B (en) | Enterococcus faecium strain and application thereof | |
| CN114196575B (en) | Panda source streptococcus paris and application thereof | |
| CN117757672A (en) | Pediococcus acidilactici with ulcerative colitis treatment function and application 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |