CN115478031A - Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof - Google Patents
Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof Download PDFInfo
- Publication number
- CN115478031A CN115478031A CN202211187476.1A CN202211187476A CN115478031A CN 115478031 A CN115478031 A CN 115478031A CN 202211187476 A CN202211187476 A CN 202211187476A CN 115478031 A CN115478031 A CN 115478031A
- Authority
- CN
- China
- Prior art keywords
- composition
- gdmcc
- bacterium
- inflammatory bowel
- bowel disease
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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
- 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
-
- 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
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- 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
-
- 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
-
- 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
- C12R2001/145—Clostridium
-
- 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)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Virology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides bile acid metabolism bacteria for preventing and treating inflammatory bowel diseases and application thereof. The invention provides a bacterium, which comprises any one, two or three of the following strains: the preservation number is GDMCC No:62707 of Clostridium sp; the preservation number is GDMCC No:62708 of Eubacterium limosum; the preservation number is GDMCC No:62709 bacterioides ovatus. The bacteria of the invention can convert primary bile acid into secondary bile acid, and improve the inflammatory symptoms of DSS-induced ulcerative colitis mice.
Description
Technical Field
The invention relates to a bacterium for preventing and treating inflammatory bowel diseases such as ulcerative colitis and application thereof, in particular to a bile acid metabolism bacterium for preventing and treating inflammatory bowel diseases, a composition containing the bacterium and related application.
Background
Inflammatory Bowel Disease (IBD) is a chronic, recurrent inflammatory disease of the intestinal tract, and patients often have persistent ulcerations at the mucosa of the small and large intestine, largely divided into two types, ulcerative colitis and Crohn's Disease (CD). Possible causes of IBD include: genetic predisposition, environmental factors such as intestinal flora, social behaviors such as smoking and diet, and an increased risk of IBD in addition to early exposure to antibiotics in children. The relative prevalence of IBD varies greatly from geographic region to geographic region, with similar but distinct pathological features. IBD has high recurrence, long course of disease, difficult cure and poor prognosis, and no medicine or treatment method for radical cure exists at present.
The pathophysiological mechanisms of IBD are not well understood and usually occur after the immune system has an exaggerated immune response to the normal intestinal flora, triggering a series of inflammatory events that destroy and destroy the intestinal wall. The intestinal mucosa barrier separates the symbiotic flora from the intestinal epithelium, immune cells at the mucosa inhibit the accumulation and translocation of the symbiotic flora, and some IBD-related gene mutations can cause the mucosal barrier or the immune response to be damaged, thereby causing the imbalance of the intestinal flora and the immune balance. To date, there are a number of reports in the literature relating IBD to dysbacteriosis, and the mechanisms of influence of changes in flora on intestinal inflammation include: regulating Treg cells and other immune cells, regulating proinflammatory and inflammation-inhibiting factors, directly invading intestinal epithelial cells and the like. In the intestine, as the largest immune organ, studies on IBD are currently conducted on polypyro short-chain fatty acids, while bile acids are mostly considered to be related to metabolic diseases such as diabetes and fatty liver, and the relationship between IBD and bile acids is rarely studied.
Similar solutions exist today including: honda et al screened 17 bacteria that modulate immune response by specific media for treatment of IBD; daniel van et al designed two synthetic bacterial populations, GUT-103 and GUT-108, for the treatment of IBD by the biogenetic analysis of a commercial strain pool.
The strains of synthetic flora which have been studied in the past are chloroform-tolerant strains obtained by adding chloroform to a culture medium, and are not rationally designed. In addition, the strains screened based on the belief analysis also lack in vitro metabolic assay validation for the metabolites of the intestinal flora of interest.
Disclosure of Invention
An object of the present invention is to provide bacteria that can prevent and treat inflammatory bowel disease.
Another object of the present invention is to provide the use of the selected bacteria.
In the research of the inventor, based on human microbial array data, aiming at the phenomenon that the excrement of an IBD patient lacks secondary bile acid, bacteria, particularly synthetic bacteria, which can metabolize combined primary bile acid into secondary bile acid are screened based on the combination of biogenic analysis and in-vitro metabolism detection, and the designed bacteria are further perfused into a DSS-induced mouse model, so that the improvement condition of the bacteria on the enteritis index of a mouse is verified, the bacteria are proved to be bile acid metabolism bacteria, and the bacteria are the synthetic bacteria which can improve IBD symptoms.
Specifically, the invention provides three strains capable of metabolizing conjugated primary bile acid into secondary bile acid, which are respectively named as Clostridium sp.DA266, eubacterium limosum DA510 and Bacteroides ovatus DA668. All three strains have been deposited, among them:
strain Clostridium sp.da266, date of deposit: 18 months 08 in 2022; the preservation unit: guangdong province culture Collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070; the preservation number is: GDMCC No. 62707; and (3) classification and naming: clostridium sp. The strain Clostridium sp.da266 of the present invention is also referred to as Clostridium sp.gdmcc-62707.
Strain Eubacterium limosum DA510, date of deposit: 18 months 08 in 2022; the preservation unit: guangdong province culture Collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070; the preservation number is: GDMCC No. 62708; and (3) classification and naming: eubacterium limosum. The strain of the invention Eubacterium limosum DA510 is also known as Eubacterium limosum GDMCC-62708.
Strain Bacteroides ovatus DA668, deposit date: 18 months 08 in 2022; the preservation unit: guangdong province culture Collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070; the preservation number is: GDMCC No. 62709; and (3) classification and naming: bacteroides ovatus. The strain Bacteroides ovatus DA668 of the present invention is also known as Bacteroides ovatus GDMCC-62709.
The research of the invention shows that any one or more of the strains Clostridium sp.da266, eubacterium limosum DA510 and Bacteroides ovatus DA668 of the invention has the capability of metabolizing conjugated primary bile acid into secondary bile acid, and can improve the symptoms of IBD. All three of the bacteria of the present invention may be referred to as bile acid metabolizing bacteria.
Thus, in one aspect, the invention provides a bacterium comprising any one, two or three of the following strains:
the preservation number is GDMCC No:62707 of Clostridium sp;
the preservation number is GDMCC No:62708 of Eubacterium limosum;
the preservation number is GDMCC No:62709 bacteriodes ovatus.
In other words, the bacterium of the present invention may be a pure culture of a single bacterium, or may be a group including two or more kinds of the bacterium.
According to a particular embodiment of the invention, the bacterium of the invention comprises the bacterium deposited under the accession number GDMCC No:62707 and having a deposit number GDMCC No:62708 of Eubacterium limosum.
According to a particular embodiment of the invention, the bacterium of the invention comprises the following deposited strain with the deposit number GDMCC No:62708 of Eubacterium limosum and having a deposit number GDMCC No:62709 bacteriodes ovatus.
According to a particular embodiment of the invention, the bacterium of the invention comprises the bacterium deposited under the accession number GDMCC No:62707 and having a deposit number GDMCC No:62709 bacteriodes ovatus.
According to a particular embodiment of the invention, the bacteria of the invention comprise the bacteria with the deposit number GDMCC No:62707 and having a deposit number GDMCC No:62708 of Eubacterium limosum and having a deposit number GDMCC No:62709 bacteriodes ovatus.
According to a specific embodiment of the present invention, in the bacteria of the present invention, the deposit number is GDMCC No:62707 the Clostridium sp has an intact bai gene cluster.
According to a specific embodiment of the present invention, in the bacteria of the present invention, the deposit number is GDMCC No:62708 strain of Eubacterium limosum GDMCC-62708 has the enzyme 7-. Alpha./. Beta. HSDH.
According to a particular embodiment of the invention, the bacterium of the invention wherein the Bacteroides ovatus GDMCC-62709 strain has a BSH enzyme.
According to a specific embodiment of the present invention, when two or three strains are included in the bacterium of the present invention, the quantitative ratio between the strains may be 0.01 to 100:0.01 to 100, preferably 0.1 to 10:0.1 to 10, more preferably 0.1 to 10:1.
according to a particular embodiment of the invention, the bacterium of the invention, which may be a solid or liquid bacterium preparation. The bacteria may be prepared into the bacterial preparation in any manner available in the art. Preferably, the bacteria in the bacterial preparation are live bacteria.
In another aspect, the present invention provides a composition comprising the bacterium of the present invention (i.e., one, two or three of Clostridium sp with deposit No. GDMCC No. 62707, eubacterium limosum with deposit No. GDMCC No. 62708, and Bacteroides ovatus with deposit No. GDMCC No. 62709). The composition of the present invention has the ability to metabolically convert conjugated primary bile acids to secondary bile acids due to the inclusion of the bacteria, and can ameliorate the symptoms of IBD. The compositions of the present invention may be referred to as bile acid metabolism compositions. Preferably, the composition may further comprise other active substances having the same or advantageous function as the bacterium of the present invention (e.g. bile acid metabolism), and/or adjuvants. According to some embodiments of the invention, the composition may be a pharmaceutical composition or a food composition, which may be, for example, a health food, a food for special medical use or a therapeutic diet. The pharmaceutical composition or the food composition can also comprise conventional auxiliary materials in the field of medicines or foods.
In another aspect, the invention also provides the use of the bacterium of the invention or the composition for the preparation of a composition for the prevention and/or treatment of inflammatory bowel disease.
According to a particular embodiment of the invention, the inflammatory bowel disease is ulcerative colitis.
In another aspect, the invention also provides the use of the bacterium of the invention or the composition for preparing a composition for alleviating weight loss and/or colon shortening in an individual with inflammatory bowel disease.
In another aspect, the invention also provides the use of a bacterium of the invention or a composition of the invention in the preparation of a composition for improving bile acid metabolism in an individual with inflammatory bowel disease.
In another aspect, the invention also provides the use of a bacterium of the invention or a composition of the invention in the preparation of a composition for improving the intestinal flora of an individual with inflammatory bowel disease.
According to a specific embodiment of the present invention, the composition for preventing and/or treating inflammatory bowel disease of the present invention may be a live bacterial preparation or an inactivated bacterial preparation. Preferably, the composition for preventing and/or treating inflammatory bowel disease is an oral formulation. The composition may be a pharmaceutical composition or a food composition, for example a health food, a food for special medical use or a therapeutic diet. The pharmaceutical composition or the food composition can also comprise conventional auxiliary materials in the field of medicines or foods.
According to a particular embodiment of the invention, when two or three bacteria of the invention are included in the composition, the quantitative ratio between the bacteria may be between 0.01 and 100:0.01 to 100, preferably 0.1 to 10:0.1 to 10, more preferably 0.1 to 10:1, for example, the number of each bacterium is substantially the same.
According to a particular embodiment of the invention, the bacteria according to the invention are used at a rate of 1X 10 per day 5 CFU～1×10 13 The amount of CFU is used to prepare the composition. Preferably, the bacteria of the present invention are administered at a rate of 1X 10 per day 7 CFU～1×10 10 The amount of CFU is used to prepare the composition.
Specific experiments of the invention show that one or more of the three bacteria can convert primary bile acid into secondary bile acid, and can improve the inflammatory symptoms of DSS-induced ulcerative colitis mice. It is reasonable to expect that the combined secondary metabolite short chain fatty acids, derivatives thereof and/or culture fluids of the three bacteria of the present invention also have certain applications in preventing and/or alleviating inflammatory bowel disease; the abundance and/or metabolites of the three bacterial combinations of the present invention can be used as markers for diagnosing inflammatory bowel disease. Furthermore, the technology of the invention can take intestinal flora as a target of disease and administration, discuss the molecular mechanism of improving IBD by the strain capable of metabolizing to generate the secondary bile acid based on the bile acid as a substance, systematically expound the molecular mechanism of treating ulcerative colitis by the bile acid and the strain capable of metabolizing the bile acid based on the structural change, differential gene expression and metabolic pathway regulation of animal model flora before and after the strain capable of metabolizing to generate the secondary bile acid is administered based on multi-group analysis.
Drawings
Figure 1 shows protection of bile acids and short chain fatty acids against a mouse model of enteritis.
FIGS. 2a to 2e show the results of tests for screening the bacterial population based on the bioassay and show three extracellular metabolic assays validation.
FIGS. 3a to 3g show the results of tests of the effect of three bacteria on body weight change and colon length in enteritis mice.
FIGS. 4a and 4b show the results of fecal bile acid content measurements after three bacteria had been exposed to enteritis mice.
FIGS. 5a and 5b show the results of fecal short chain fatty acid content measurements after three bacteria had been exposed to enteritis mice.
Fig. 6 shows the results of metagenomic sequencing of the effect of the three bacteria combination on intestinal flora in enteritis mice.
Microbial deposits for patent procedures:
(ii) strain Clostridium sp.DA266
The preservation date is as follows: 18 months 08 in 2022;
the preservation unit: guangdong province culture Collection of microorganisms (GDMCC);
the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070;
the preservation number is: GDMCC No. 62707;
and (3) classification and naming: clostridium sp.
(II) Strain Eubacterium limosum DA510
The preservation date is as follows: 18 months 08 in 2022;
the preservation unit is as follows: guangdong province culture Collection of microorganisms (GDMCC);
the address of the depository: china Guangzhou city, renlie Zhonlu Dazhou No. 100 college No. 59 building, guangdong province academy of sciences, microbial institute, postal code: 510070;
the preservation number is: GDMCC No. 62708;
and (3) classification and naming: eubacterium limosum.
(III) Strain bacterioides ovatus DA668
The preservation date is as follows: 18 months 08 in 2022;
the preservation unit is as follows: guangdong province culture Collection of microorganisms (GDMCC);
the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070;
the preservation number is as follows: GDMCC No. 62709;
and (3) classification and naming: bacteroides ovatus.
Detailed Description
In order to clearly understand the technical features, objects and advantages of the present invention, the following detailed description of the technical solutions of the present invention is made with reference to the specific embodiments and the accompanying drawings, which are understood to be merely illustrative of the present invention and not limiting the scope of the present invention. Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art. The method operations not specifically mentioned in the examples were carried out according to the conventional operations of the prior art or the operations suggested by the manufacturer's specifications.
Example 1 protection of Compounds on murine enteritis model
This example is based primarily on the protection of compounds against a murine model of enteritis. After adaptive feeding, mice were divided into 2 groups, respectively the double acid and SCFA group and the DSS group, and modeling was started 10 days after preventive administration in advance, i.e., drinking water was changed to 2% DSS water, and mice were sacrificed every day on the seventh day of mouse weight modeling. The results are shown in FIG. 1, and the body weight loss of mice in the model group of bile acid and SCFA is significantly reduced at day seven compared with that in the control group. The combination of the bile acid and the SCFA has better protection on enteritis of mice.
Example 2 Strain screening
This example is based primarily on the combination of a belief-generating assay and in vitro metabolic assays to screen for synthetic flora that metabolize conjugated primary bile acids to secondary bile acids.
The strain library screened by the invention is 48 strains obtained by separation culture in a healthy human intestinal fecal sample in the early stage of the laboratory, all the strains are subjected to monoclonal purification, and the species of the strains are determined by 16S rna sequencing identification (figure 2 a);
and (3) letter generation analysis: screening strains with 7-alpha dehydroxylase, BSH enzyme and 7-alpha/beta HSDH enzyme, wherein the screened database is derived from strains separated and purified from healthy human intestinal fecal samples in the early stage of the laboratory, and is subjected to whole genome sequencing, and the analysis result is as follows, two strains with complete bai gene clusters are used for detecting the in vitro metabolic capacity, wherein one strain is named as Clostridium sp.DA266; the strain with 7 alpha/beta only has one strain, and is named as Eubacterium limosum DA510 in the invention; BSH exists in a plurality of strains, and based on the report of better enteritis treatment effect of Bacteroides ovatus in the literature, a strain named as Bacteroides ovatus DA668 is finally selected in the invention. All three strains have been deposited, among them: strain Clostridium sp.da266, date of deposit: 18 months 08 in 2022; the preservation unit: guangdong province culture collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070; the preservation number is: GDMCC No. 62707; and (3) classification and naming: clostridium sp. The strain of the invention Clostridium sp.da266 is also referred to as Clostridium sp: GDMCC-62707. Strain Eubacterium limosum DA510, date of deposit: 18 months 08 in 2022; the preservation unit is as follows: guangdong province culture Collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, pieli Zhonglu 100 Dazhou 59 building, guangdong province academy of sciences, microbial research institute, zip code: 510070; the preservation number is as follows: GDMCC No. 62708; and (3) classification and naming: eubacterium limosum. The strain of the invention Eubacterium limosum DA510 is also known as Eubacterium limosum GDMCC-62708. Strain Bacteroides ovatus DA668, deposit date: 18 months 08 in 2022; the preservation unit: guangdong province culture Collection of microorganisms (GDMCC); the address of the depository: china Guangzhou city, renlie Zhonlu Dazhou No. 100 college No. 59 building, guangdong province academy of sciences, microbial institute, postal code: 510070; the preservation number is: GDMCC No. 62709; and (3) classification and naming: bacteroides ovatus. The strain Bacteroides ovatus DA668 of the present invention is also known as Bacteroides ovatus GDMCC-62709. The invention selects three strains of Clostridium sp GDMCC-62707, eubacterium limosum.
In vitro metabolic assays verified whether the designed strains screened had the enzymes mentioned: (1) resuscitating the screened strain by using a brain-heart leachate BHI culture medium, then transferring the resuscitated bacterial liquid into a new culture medium containing 50 mu mol of primary conjugated bile acid TCDCA/GCDCA, culturing for 48 hours, and collecting a culture solution; (2) adding equal volume of ethyl acetate into 200 mu L of the collected culture solution, carrying out vigorous vortexing for 15 minutes, centrifuging at 10000rpm for 5 minutes, sucking a supernatant ethyl acetate layer into a new centrifugal tube of 1.5 ml, airing, dissolving with 200 mu L of 50% methanol, centrifuging at 10000rpm for 10 minutes, filtering with a 0.22um filter membrane, collecting filtrate in a liquid phase sample bottle, and storing in a refrigerator of-80 ℃. (3) The results of liquid chromatography mass spectrometry are shown in fig. 2b, fig. 2c, fig. 2d, fig. 2e, and the substrate TCDCA/GCDCA is metabolically transformed by co-culture of 3 bacteria to produce secondary bile acids UDCA and LCA.
Example 3 protection of the combination bacterium SBE against DSS-induced enteritis model in mice
After adaptive feeding, 48 9-week-old C57 mice were randomly divided into 6 groups of 8, one (C.scintillans, eubacterium, bacteroides ovalis), two (C.deltoides, three) and one control group. The total bacterial load of all the bacteria-feeding groups is the same, namely, the single bacteria group is gavaged by 1 multiplied by 10 every day 9 CFU, two bacteria combination group Clostridium sp GDMCC-62707, eubacterium limosum GDMCC-62708, respectively filled with 5X 10 of water 8 CFU, three bacteria combination group Clostridium sp GDMCC-62707, eubacterium limosum GDMCC-62708, bacteria broth inside GDMCC-62709, 3.3X 10/day respectively 8 CFU, a blank culture medium with the same volume as the control group is perfused for 10 days continuously, and the feces of each group of mice are collected on the 10 th day and immediately put into a liquid nitrogen tank for laboratory preservation at-80 ℃. Then, the drinking water of the mice was changed to water containing 2% DSS, DSS-induced mice enteritis model, mice were allowed to drink water freely, the gavage was continued, and the weight of the mice was recorded daily, model building was continued for 7 days, mouse feces were collected from each group, mice were sacrificed, serum and colon were collected, the length of the colon, feces, colon and blood were recorded and stored in a refrigerator at-80 ℃. The weight change and the colon length of the mice are shown in figures 3a to 3g, and the three-bacterium combination has better effects on alleviating the weight loss and the colon shortening of the mice than the single-bacterium group and the two-bacterium combination.
Example 4 Effect of three bacteria combinations on fecal bile acids and short chain fatty acids in enteritis mice
From the results (fig. 4a and 4 b) of the determination of bile acid content in feces of mice (feeding protocol same as example 3), it can be seen that the contents of UDCA and LCA in the secondary bile acid DCA of the three-bacterium gavage combination 3strains group are increased compared with the control group, and the result is consistent with the conclusion that 3strains can metabolize and convert the conjugated primary bile acid into the secondary bile acid in the in vitro test of the invention. In addition, the amount of short chain fatty acids propionic acid and butyric acid was also increased in the 3-bacterium gavage group (fig. 5a and 5 b).
Example 5 Effect of the combination of three bacteria on intestinal flora of enteritis mice
From the results of sequencing of the fecal metagenome of mice (feeding protocol same as example 3) (fig. 6), it can be seen that the abundance of some bacteria in the intestinal flora was changed by gavage with three bacteria compared to the DSS-control group.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including various technical features being combined in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (10)
1. A bacterium comprising any one, two or three of the following strains:
the preservation number is GDMCC No:62707 Clostridium sp;
the preservation number is GDMCC No:62708 of Eubacterium limosum;
the preservation number is GDMCC No:62709 bacteriodes ovatus.
2. The bacterium of claim 1, wherein the deposit number is GDMCC No:62707 Clostridium sp. having the entire bai gene cluster;
the preservation number is GDMCC No:62708 strain GDMCC-62708 has 7-alpha/beta HSDH enzyme; and/or
The Bacteroides ovatus GDMCC-62709 strain has BSH enzyme.
3. The bacterium according to claim 1 or 2, which is a solid or liquid bacterium preparation.
4. A composition comprising the bacterium of any one of claims 1-3; preferably, the composition is a pharmaceutical composition or a food composition.
5. Use of a bacterium according to any one of claims 1 to 3 or a composition according to claim 4 for the preparation of a composition for the prevention and/or treatment of inflammatory bowel disease;
preferably, the inflammatory bowel disease is ulcerative colitis.
6. Use of a bacterium according to any one of claims 1 to 3 or a composition according to claim 4 in the manufacture of a composition for use in the alleviation of weight loss and/or colon shortening in an individual with inflammatory bowel disease.
7. Use of the bacterium of any one of claims 1-3 or the composition of claim 4 in the preparation of a composition for improving bile acid metabolism in an individual with inflammatory bowel disease.
8. Use of a bacterium according to any one of claims 1 to 3 or a composition according to claim 4 for the preparation of a composition for improving the intestinal flora of an individual with inflammatory bowel disease.
9. The use according to any one of claims 5 to 8, wherein the composition for the prevention and/or treatment of inflammatory bowel disease is a live or inactivated bacterial preparation; preferably, the composition for preventing and/or treating inflammatory bowel disease is an oral formulation.
10. Use according to any one of claims 5 to 9, wherein the bacteria of any one of claims 1 to 3 are administered at a rate of 1 x 10/day 5 CFU～1×10 13 CFU is preferably 1 × 10 7 CFU～1×10 10 The amount of CFU is used to prepare the composition.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211187476.1A CN115478031A (en) | 2022-09-28 | 2022-09-28 | Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof |
| PCT/CN2022/139762 WO2024066079A1 (en) | 2022-09-28 | 2022-12-16 | Bile acid metabolizing bacterium for preventing and treating inflammatory bowel disease and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211187476.1A CN115478031A (en) | 2022-09-28 | 2022-09-28 | Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115478031A true CN115478031A (en) | 2022-12-16 |
Family
ID=84394728
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211187476.1A Pending CN115478031A (en) | 2022-09-28 | 2022-09-28 | Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115478031A (en) |
| WO (1) | WO2024066079A1 (en) |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018112739A1 (en) * | 2016-12-20 | 2018-06-28 | 深圳华大基因研究院 | Bifidobacterium pseudocatenulatum, culture method therefor and application thereof |
| CA3095402A1 (en) * | 2018-03-29 | 2019-10-03 | Seres Therapeutics, Inc. | Compositions and methods for treating inflammatory bowel diseases |
| IL271775A (en) * | 2019-12-31 | 2021-06-30 | Biomica Ltd | Microbial consortium and uses thereof |
| CN116056717A (en) * | 2020-07-13 | 2023-05-02 | 亚洲大学校产学协力团 | Composition for preventing or treating inflammatory bowel disease |
| KR102470808B1 (en) * | 2020-11-02 | 2022-11-28 | 주식회사 바이오미 | Composition for the preventation or treatment of inflammatory bowel disease |
-
2022
- 2022-09-28 CN CN202211187476.1A patent/CN115478031A/en active Pending
- 2022-12-16 WO PCT/CN2022/139762 patent/WO2024066079A1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| WO2024066079A1 (en) | 2024-04-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2022033885A (en) | Designed bacterial compositions | |
| US10246677B2 (en) | Devices, systems and methods for the production of humanized gut commensal microbiota | |
| US20220133814A1 (en) | Faecalibacterium prausnitzii and desulfovibrio piger for use in the treatment or prevention of diabetes and bowel diseases | |
| Duncan et al. | Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system | |
| Fukui | Gut microbiota and host reaction in liver diseases | |
| Li et al. | Altered short chain fatty acid profiles induced by dietary fiber intervention regulate AMPK levels and intestinal homeostasis | |
| Kanauchi et al. | Eubacterium limosum ameliorates experimental colitis and metabolite of microbe attenuates colonic inflammatory action with increase of mucosal integrity | |
| RU2629806C2 (en) | Bacterial strains, belonging to bifidobacterium genus for application in hypercholesterinemia treatment | |
| Dong et al. | 16S rDNA full-length assembly sequencing technology analysis of intestinal microbiome in polycystic ovary syndrome | |
| CN111212655A (en) | Compositions and methods for treating cholestatic diseases | |
| CN115216422B (en) | Lactobacillus rhamnosus and application thereof | |
| Zhang et al. | Polysaccharides from edible brown seaweed Undaria pinnatifida are effective against high-fat diet-induced obesity in mice through the modulation of intestinal microecology | |
| CN115011532B (en) | Lactobacillus paracasei JY062 preparation, and preparation method and application thereof | |
| CN114159478A (en) | Bacterial powder composition for relieving inflammatory colitis and preparation method and application thereof | |
| CN113897302A (en) | Bifidobacterium capable of relieving colitis and application thereof | |
| WO2018134361A1 (en) | Autologous fecal sample for use in the treatment of microbial dysbiosis | |
| Askari et al. | The Role of Gut Microbiota in Inflammatory Bowel Disease-Current State of the Art | |
| CN115478031A (en) | Bile acid metabolism bacterium for preventing and treating inflammatory bowel disease and application thereof | |
| Li et al. | The bioavailability of glycyrrhizinic acid was enhanced by probiotic Lactobacillus rhamnosus R0011 supplementation in liver fibrosis rats | |
| CN108159090A (en) | Purposes of the lucidum spore powder dietary fiber extract in treatment and/or prevention intestinal bacilli illness relevant disease preparation is prepared | |
| WO2021173895A1 (en) | Analysis of microbiome for diagnosis and treating of urinary stone disease | |
| WO2022164422A1 (en) | Methods and compositions for precision release of probiotics to improve human health | |
| CN114606166B (en) | Bacteroides xylanisolvens and application thereof in preparation of products for treating or preventing alcoholic liver disease | |
| CN116555126B (en) | Composition containing probiotics and having enteritis treatment effect and application thereof | |
| RU2773327C2 (en) | Developed bacterial compositions |
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 |