CN115400153A - Application of bacteroides in preparation of product for preventing or treating nicotine-related metabolic diseases, related product and culture method - Google Patents

Application of bacteroides in preparation of product for preventing or treating nicotine-related metabolic diseases, related product and culture method Download PDF

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CN115400153A
CN115400153A CN202211202491.9A CN202211202491A CN115400153A CN 115400153 A CN115400153 A CN 115400153A CN 202211202491 A CN202211202491 A CN 202211202491A CN 115400153 A CN115400153 A CN 115400153A
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bacteroides
product
nicotine
related metabolic
preventing
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姜长涛
陈博
曾广易
汪锴
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Peking University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • 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
    • 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
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • 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
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • 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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
    • A61P25/34—Tobacco-abuse
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • 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
    • 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

Abstract

The invention discloses application of bacteroides in preparation of a product for preventing and/or treating nicotine-related metabolic diseases, application of bacteroides in preparation of a product for preventing or treating nicotine-related metabolic diseases, a related product and a bacteroides culture method; the product for preventing or treating nicotine-related metabolic diseases comprises medicines, foods, beverages and feeds, can effectively degrade nicotine, and remarkably improves nicotine-aggravated symptoms of blood sugar, blood fat disorder and non-alcoholic fatty liver diseases.

Description

Application of bacteroides in preparation of product for preventing or treating nicotine-related metabolic diseases, related product and culture method
Technical Field
The invention relates to the field of microbiology, in particular to application of Bacteroides spp (Bacteroides spp.) in preparing products for preventing or treating nicotine-related metabolic diseases, related products and a culture method.
Background
Nicotine is a bitter alkaloid compound, and is naturally present in a large number of solanaceae plants, including various plants such as tobacco. Nicotine is both addictive and toxic, and its ingestion or inhalation (or other mode of use) is associated with a variety of metabolic disorders. The most common source of nicotine in the body is smoking. Smoking is a global health care problem and also a major cause of death prevention. According to the world health organization estimates that there are currently 11 billion smokers worldwide and nearly six million deaths per year are associated with tobacco, reckoned with the current trend, and by 2030, the number of deaths from smoking will reach about eight million people per year.
Metabolic diseases are a general term for diseases caused by metabolic disorders in vivo, and include hyperglycemia, hyperlipidemia, arteriosclerosis, hypertension, non-alcoholic fatty liver disease, and the like. Various clinical studies show that nicotine intake can induce/aggravate various metabolic diseases such as insulin resistance, hyperglycemia, hyperlipidemia, non-alcoholic fatty liver disease and the like. The method can effectively metabolize nicotine and degrade the content of nicotine in vivo, and is an important strategy for relieving nicotine-induced metabolic diseases.
The development and exploration of probiotics are carried out for many years from development to clinical application, the categories and action mechanisms of the probiotics are gradually clear, and the probiotics are widely applied in clinic and relate to organ protection, immunity, bacteriostasis, blood fat reduction, blood sugar reduction and the like. Bacteroides are an important group of gram-positive bacteria in the intestinal tract, and some studies have shown that their content in the intestinal tract is reduced in patients with diabetes, systemic inflammation and corresponding animal models of the disease. However, bacteroides, such as Bacteroides xylanisolvens (Bacteroides xylanisolvens), bacteroides dorsalis (Bacteroides dorei), bacteroides ovatus (Bacteroides ovatus), bacteroides fragilis (Bacteroides fragilis), bacteroides vulgatus (Bacteroides vulgatus), and the like, have not been studied and found to have a nicotine-degrading effect, and metabolic diseases such as nicotine-induced diabetes and non-alcoholic fatty liver have not been reported as to whether or not the bacteria can alleviate or treat nicotine-induced diabetes. We found that Bacteroides can effectively degrade nicotine in vivo and in vitro, thereby improving the metabolic disease process induced by the nicotianine.
Disclosure of Invention
Aiming at the limitation that the existing targeted nicotine degradation strategy improves nicotine-related metabolic diseases, the invention provides application of bacteroides in preparing a product for preventing or treating nicotine-related metabolic diseases, a related product and a culture method.
In a first aspect of the invention, there is provided the use of a bacteroides in the manufacture of a product for the prevention or treatment of a nicotine-related metabolic disorder;
further, in the present invention, the product is any one of: drugs, foods, beverages, feeds;
further, in the present invention, the bacteroides is one or more of: bacteroides xylanisolvens, bacteroides dorferi, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus; preferably, the Bacteroides xylanisolvens is Bacteroides xylanisolvens DSM18836, the Bacteroides dorei is Bacteroides dorei DSM17855, and the Bacteroides ovatus is Bacteroides ovatus
Figure BDA0003872910420000021
8483 the Bacteroides fragilis is Bacteroides fragilis
Figure BDA0003872910420000022
25285 Bacteroides vulgatus, bacteroides vulgatus
Figure BDA0003872910420000023
8482。
Further, in the present invention, the nicotine-related metabolic diseases include hyperglycemia, hyperlipidemia, and non-alcoholic fatty liver disease caused/aggravated by nicotine usage.
In a second aspect of the invention, there is provided a product for preventing or treating nicotine-related metabolic disorders, the product comprising an effective amount of bacteroides.
Further, in the product of the invention, the bacteroides is one or more of: bacteroides xylanisolvens, bacteroides dorferi, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus; preferably, jie Muju Bacteroides saccharolyticus is Bacteroides xylansolvens DSM18836, bacteroides dorei is Bacteroides dorei DSM17855, and Bacteroides ovatus is Bacteroides ovatus
Figure BDA0003872910420000024
8483 the Bacteroides fragilis is Bacteroides fragilis
Figure BDA0003872910420000025
25285, bacteroides vulgatus is Bacteroides vulgatus
Figure BDA0003872910420000026
8482。
Further, in the product of the invention, the content of the bacteroides is 1 × 10-1 × 10 20 cfu/mL, preferably 1X 10 7 -1×10 9 cfu/mL。
Further, in the present invention, the product is any one of: medicine, food, beverage, and feed.
Optionally, in the present invention, the food product comprises an effective amount of bacteroides and/or metabolites thereof, and the balance food acceptable carriers;
alternatively, in the present invention, the type of the food may be a solid, milk, solution product, powder product, or suspension product;
optionally, in the present invention, the medicament further comprises a pharmaceutically acceptable excipient;
further, the pharmaceutically acceptable adjuvant may be at least one selected from the group consisting of a carrier, an excipient, a diluent, a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor;
further, the pharmaceutically acceptable excipient may be at least one selected from lactose, glucose, sucrose, sorbitol, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone (PVP), cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate or mineral oil;
optionally, in the present invention, the dosage form of the drug is selected from any one of the following: granules, capsules, tablets, powders, oral liquids, suspensions, and emulsions.
The drug of the present invention can be administered in any form of pharmaceutical tablet, injection or capsule, and further comprises excipients, pharmaceutically acceptable vehicles and carriers, which can be selected according to the administration route.
Further, the medicament of the present invention may further comprise a lubricant, a wetting agent, an emulsifier, a suspoemulsion stabilizer, a preservative, a sweetener, a flavor, and the like.
The medicament of the present invention can be produced in enteric-coated tablets for oral use by various known methods, so that the active ingredient of the pharmaceutical composition, i.e., the microorganism, can pass through the stomach smoothly without being destroyed by gastric acid; "enteric coating" includes all coatings which are approved for use with conventional drugs and which are not degraded by gastric acid but which are sufficiently decomposed in the small intestine to rapidly release the microorganisms of the present invention.
The enteric coating of the invention is capable of being maintained at 36-38 ℃ for more than 2 hours in synthetic gastric acid, e.g. HCl solution at pH =1, and preferably disintegrates within 1.0 hour in synthetic intestinal fluid, e.g. buffer at pH = 7.0.
The enteric coating of the invention is coated at about 16-30mg, preferably 16-25mg, more preferably 16-20mg per tablet. The thickness of the casing of the invention is 5-100 μm, and the preferred thickness is 20-80 μm. The enteric coating composition itself is known to be a conventional polymer.
In a third aspect of the present invention, there is provided a bacteroides culture method, comprising: inoculating Bacteroides onto the slant under anaerobic condition, and growing in the culture medium on the slant at 10-40 deg.C for 2-3 days; selecting thallus, inoculating the thallus into a culture solution, and culturing in the culture solution at the temperature of 10-40 ℃ for 2-3 days, wherein the culture medium comprises 10-30g/L of a carbon source, 6-15g/L of a nitrogen source, 10-20g/L of agar and 0.2-1g/L of an inorganic salt composition, and the pH value is 3.0-7.0; the culture solution comprises 10-30g/L of carbon source, 6-15g/L of nitrogen source, 1-3g/L of agar and 0.2-1g/L of inorganic salt composition, and the pH value is 3.0-7.0. The method significantly improves the production of Bacteroides.
Further, the culture medium comprises 10, 20, 30g/L carbon source, 6, 9, 12, 15g/L nitrogen source, 10, 15, 20g/L agar, 0.2, 0.5, 0.8, 1g/L inorganic salt composition, pH value is 3.0, 5.0, 7.0;
further, the culture solution contains 10, 20, 30g/L carbon source, 6, 9, 12, 15g/L nitrogen source, 1, 2, 3g/L agar, 0.2, 0.5, 0.8, 1g/L inorganic salt composition, and has pH value of 3.0, 5.0, 7.0. Optionally, in the method of the present invention, the carbon source is fructose, glucose, or sucrose.
Optionally, in the method of the present invention, the nitrogen source is corn steep liquor, yeast extract, or corn meal.
Optionally, in the method of the present invention, the inorganic salt composition comprises: potassium dihydrogen phosphate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, anhydrous calcium chloride and sodium chloride.
In a fourth aspect of the invention, there is provided the use of a bacteroides in the prevention and/or treatment of nicotine related metabolic disorders.
Further, the simulationThe bacillus is one or more of: bacteroides xylanisolvens, bacteroides dorsalis, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus; preferably, the Bacteroides xylanisolvens is Bacteroides xylanisolvens DSM18836, the Bacteroides dorei is Bacteroides dorei DSM17855, and the Bacteroides ovatus is Bacteroides ovatus
Figure BDA0003872910420000041
8483 the Bacteroides fragilis is Bacteroides fragilis
Figure BDA0003872910420000042
25285, bacteroides vulgatus is Bacteroides vulgatus
Figure BDA0003872910420000043
8482。
In a fifth aspect of the present invention, there is provided a method for treating or preventing obesity-related diseases, which comprises administering the product of the present invention to a target subject (human or animal), wherein nicotine is effectively degraded, and the nicotine-induced blood sugar, blood lipid disorders and non-alcoholic fatty liver disease symptoms are significantly improved, and the product can be used for the preparation of a medicament for treating nicotine-induced/aggravated diabetes, blood lipid disorders and non-alcoholic fatty liver disease.
Advantages of the present invention include, but are not limited to, the following:
1. the degradation effect of the bacteroides on nicotine is found in vitro experiments and animal experiments for the first time;
2. bacteroides were found to be effective in preventing and treating nicotine-aggravated metabolic disorders, and organisms that could benefit include, but are not limited to, humans, domestic animals, farm animals, and the like;
3. in order to prepare a large amount of experimental materials and products, the inventors conducted comparative culture experiments and provided a method for culturing bacteroides with high growth capacity;
4. there are provided various forms of products, such as drugs, foods, drinks, feeds, which can effectively prevent and treat nicotine-aggravated metabolic diseases.
Detailed Description
The inventors have found that Bacteroides spp has an action of degrading nicotine in vitro experiments and mouse experiments and an action of effectively preventing and treating nicotine-aggravated metabolic diseases, and that an active composition containing Bacteroides spp is fed to an experimental subject to improve insulin resistance, reduce blood sugar, improve non-alcoholic fatty liver diseases and effectively reduce diabetes, hyperlipidemia and non-alcoholic fatty liver diseases in a nicotine-aggravated metabolic disease mouse model.
As used herein, the term "comprising" means that the various ingredients can be applied together in a mixture or composition of the invention. Thus, the terms "consisting essentially of …" and "consisting of …" are encompassed by the term "comprising".
The invention provides application of Bacteroides (Bacteroides spp.) in treating and preventing nicotine-aggravated metabolic diseases and application of Bacteroides in preparing products for preventing or treating nicotine-related metabolic diseases. High lipid-induced mice are given nicotine stimulation to cause nicotine-induced metabolic disease models, bacteroides Xylanisolvens (BX), bacteroides Dorei (BD), bacteroides Ovatus (BO), bacteroides Fragilis (BF), bacteroides Vulgare (BV) have the ability to improve insulin resistance, hyperlipidemia and non-alcoholic fatty liver in nicotine-aggravated metabolic disease model animals. According to the embodiment of the invention, the nicotine-aggravated metabolic disease model mice are treated by the strains BX, BD, BO, BF, BV, and the insulin sensitivity, the blood lipid level and the liver are kept normal compared with the untreated control group. Therefore, the strain can be used for preventing and treating metabolic diseases with nicotine aggravation.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Among the strains used in the examples described below, bacteroides xylanisolvens and Bacteroides dorferi were purchased from the DSMZ culture Collection, germany, and the strain numbers are Bacteroides xylanisolvens (Bacteroides xylansolvens DSM 18836) and Bacteroides dorferi (Bacteroides dorei DSM 17855), respectively; the remaining strains were purchased from American Type Culture Collection (ATCC) and the strain number Bacteroides ovatus (Bacteroides ovatus)
Figure BDA0003872910420000051
8483 Bacteroides fragilis (Bacteroides fragilis), bacteroides fragilis (Bacteroides fragilis)
Figure BDA0003872910420000052
25285 Bacteroides vulgatus (Bacteroides vulgatus)
Figure BDA0003872910420000053
8482)。
Example 1 in vitro degradation of nicotine by Bacteroides spp
The material and the method are as follows:
the tested strains are Bacteroides Xylanisolvens (BX), bacteroides Dorsalis (BD), bacteroides Ovatus (BO), bacteroides Fragilis (BF), bacteroides fragilis (BV), and are anaerobically cultured in brain heart infusion broth culture medium (BHI) with a final concentration of 1mg/mL, and the strain (10) is added respectively 7 ～10 9 CFU/mL) is subjected to anaerobic culture in a constant-temperature incubator at 37 ℃ for 48 hours, 1mL of bacterial liquid is taken, ultrasonic crushing is carried out for 5min, centrifugation is carried out at room temperature for 5000g for 5min, and supernatant is taken to detect the content of nicotine by utilizing a mass spectrometry method.
As a result: the above test strains all degraded nicotine in the medium to different degrees (table 1), demonstrating the effective nicotine degradation by bacteroides.
TABLE 1 in vitro degradation of nicotine by BX, BD, BO, BF, BV
Grouping Nicotine content (mg/mL)
Blank space 0.986±0.121
BX 0.212±0.023
BD 0.392±0.044
BO 0.304±0.027
BF 0.426±0.034
BV 0.452±0.121
Example 2 Effect of Bacteroides on non-alcoholic fatty liver disease
Materials: the tested strains were Bacteroides Xylanisolvens (BX), bacteroides Dorsalis (BD), bacteroides Ovatus (BO), bacteroides Fragilis (BF), bacteroides Vulgatus (BV), and the amount of the gastric lavage bacteria was determined to be 0.1mL/10g body weight, and the concentration was 1X 10 9 cfu/mL, once daily. The culture medium is cultured in advance, activated every week to ensure freshness, and the concentration is measured.
The C57BL/6J mice were purchased from Jiangsu Jiejiegaokang Biotechnology GmbH, temperature 20-24 ℃, constant humidity 50-60%, light illumination 12 hours (8-00-20), sound insulation, free food intake, drinking water, and experiments were performed after adapting to the environment for one week, and fed with high fructose high cholesterol feed (HFHCD, research Diet, D09100310), while drinking water contained 23.1g/L fructose and 18.6g/L glucose. The high fructose and high cholesterol feed can induce liver cell steatosis, liver inflammation and liver injury, and simultaneously, the blood lipid level is increased and insulin resistance is induced, so the method is a common modeling method of rodent fatty liver. Nicotine (final concentration 10 ug/mL) was administered as free-water. Total Cholesterol (TC) kit (batch No. 20131112), triglyceride (TG) kit (batch No. 20131226), glutamic-oxaloacetic transaminase (ALT kit (batch No. 20131006) purchased from Nanjing institute of bioengineering
The method comprises the following steps: the male C57BL/6J mice are 8-10 weeks, and have the body weight of 24-28g, and are averagely divided according to the body weight conditions: (1) control group, (2) nicotine-treated group (Nic), (3) nicotine (Nic) + BX 1X 10 9 cfu/mL, (4) Nic) + BD 1X 10 9 cfu/mL, (5) Nic) + BO 1 × 10 9 cfu/mL, (6) Nic) + BF 1X 10 9 cfu/mL, (7) Nic) + BV 1X 10 9 cfu/mL, 8 per group, was treated continuously for 20 weeks. The model control group was given an equal amount of anaerobic PBS. After treating the mice, taking plasma and liver tissues, centrifuging the plasma at 4 ℃ and 3000rpm for 15min, and measuring ALT and AST; calculating the liver/body weight ratio of the liver, and detecting the content of TC and TG.
The experimental results show (table 2), compared with the control group, the liver index of the nicotine-treated group is obviously increased, and the liver index of the non-alcoholic fatty liver disease mouse can be obviously reduced after the bacteroides is given. Meanwhile, TC and TG of the liver after nicotine treatment are remarkably increased, which prompts that the nicotine treatment causes obvious liver lipid disorder, and compared with a nitudin treatment group, the liver lipid disorder of a mouse with non-alcoholic fatty liver disease can be reduced after five bacteroides are given, and simultaneously, the liver function can be improved, and the content of ALT and AST in blood plasma can be reduced.
TABLE 2 influence of Bacteroides strains (BX, BD, BO, BF, BV) on liver-related indices of nicotine-stimulated non-alcoholic fatty liver model mice
Figure BDA0003872910420000071
Example 3 Effect of Bacteroides (Bacteroides) on blood glucose and blood lipid levels in Nicotine-aggravated diabetic model mice
Materials: the tested strains were Bacteroides Xylanisolvens (BX), bacteroides Dorei (BD), bacteroides Ovatus (BO), bacteroides Fragilis (BF), bacteroides Vulgare (BV), and the amount of the gastric lavage bacteria was determined to be 0.10mL/10g body weight and the concentration was 1X 10 9 cfu/mL, once daily. The culture broth was cultured in advance, activated every week to ensure freshness, and the concentrations were measured separately.
C57BL/6J mice were purchased from jiangsu jiejiaokang biotechnology, inc, at 20-24 degrees celsius, at 50-60% constant humidity, irradiated for 12 hours (8. Nicotine (final concentration 10 ug/mL) was administered as free-water. Blood glucose test strips (Roche), a glucometer (Roche), a total serum cholesterol TC kit (lot number 20131112), a low-density lipoprotein LDL-C kit (lot number 20140114), a high-density lipoprotein HDL-C kit (lot number 20140413), and a triglyceride TG kit (lot number 20131226) were purchased from Nanjing, to establish a bioengineering institute.
The method comprises the following steps: male C57BL/6J mice, 6-8 weeks, 24-28g body weight, fasting blood glucose was measured, and the blood glucose and body weight were averagely grouped: (1) control group, (2) nicotine-treated group (Nic), (3) nicotine (Nic) + BX 1X 10 9 cfu/mL, (4) Nic) + BD 1X 10 9 cfu/mL, (5) Nic) + BO 1 × 10 9 cfu/mL, (6) Nic) + BF 1X 10 9 cfu/mL, (7) Nic) + BV 1X 10 9 cfu/mL, 8 per group, for 8 weeks. Blood glucose changes were measured weekly and model controls were given equal amounts of anaerobic PBS. After treating the mice, blood was collected, centrifuged at 3000rpm at 4 ℃ for 15min, and serum total cholesterol TC, low density lipoprotein L were measuredDL-C, triglyceride TG.
As a result: compared with the control group, the indexes of the nicotine treatment group are obviously different, which proves that the nicotine causes further deterioration of the mouse state, and compared with the nicotine treatment group, BX, BD, BO, BF and BV strains can reduce free diet blood sugar and fasting (4 hours) blood sugar from 3 weeks and improve the blood fat condition of mice, wherein the BX effect is most obvious, and the result is shown in a table 3,4.
TABLE 3 blood sugar Effect of Bacteroides strains (BX, BD, BO, BF, BV) on Nicotine-stimulated hyperglycemic and hyperlipidemic model mice
Figure BDA0003872910420000081
TABLE 4 influence of Bacteroides strains (BX, BD, BO, BF, BV) on blood lipids in nicotine-stimulated hyperglycemic and hyperlipidemic model mice
Figure BDA0003872910420000082
Figure BDA0003872910420000091
Example 4
Efficient culture method of bacteroides
To prepare a large number of experimental materials and products, the inventors performed comparative culture experiments:
bacteroides Xylanisolvens (BX), bacteroides Dorsalis (BD), bacteroides Ovorans (BO), bacteroides Fragilis (BF), bacteroides Vulgatus (BV) are grown in a culture medium on a slant for 2 or 3 days at 4 temperatures of 10, 20, 30 and 40 ℃ respectively; selecting bacteria, inoculating the bacteria into a culture medium, and culturing for 2 or 3 days at 4 temperatures of 10, 20, 30 and 40 ℃ in the culture medium, wherein the culture medium comprises 10, 20, 30g/L carbon source, 6, 9, 12, 15g/L nitrogen source, 10, 15, 20g/L agar, 0.2, 0.5, 0.8 and 1g/L inorganic salt composition, and has pH values of 3.0, 5.0 and 7.0; the culture solution comprises 10, 20, 30g/L carbon source, 6, 9, 12, 15g/L nitrogen source, 1, 2, 3g/L agar, 0.2, 0.5, 0.8 and 1g/L inorganic salt composition, and has pH values of 3.0, 5.0 and 7.0; wherein the carbon source is fructose, glucose or sucrose, the nitrogen source is corn steep liquor, yeast extract or corn flour, and the inorganic salt composition comprises: potassium dihydrogen phosphate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, anhydrous calcium chloride and sodium chloride.
Compared with the prior art, the yield of the bacteroides can be remarkably improved.
It is understood that within the scope of the present invention, each of the above technical features of the present invention and each of the technical features specifically described in the embodiments may be combined with each other to constitute a new or preferred technical solution. For reasons of brevity and clarity, they will not be described in detail herein.

Claims (19)

1. Use of a bacteroides in the manufacture of a product for the prevention or treatment of nicotine-related metabolic disorders.
2. Use according to claim 1, wherein the product is any one of: medicine, food, beverage, and feed.
3. The use according to claim 1, wherein the bacteroides is one or more of: bacteroides xylanolyticus, bacteroides dorsalis, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus.
4. The use of claim 1, wherein said nicotine-related metabolic disorder comprises hyperglycemia, hyperlipidemia, non-alcoholic fatty liver disease induced/exacerbated by nicotine usage.
5. A product for use in the prevention or treatment of nicotine-related metabolic disorders, comprising an effective amount of a bacteroides.
6. The product of claim 5, wherein the bacteroides is one or more of: bacteroides xylanolyticus, bacteroides dorsalis, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus.
7. The product of claim 5, wherein the Bacteroides is present in an amount of 1X 10 to 1X 10 20 cfu/mL。
8. The product of claim 5, wherein the product is any one of: medicine, food, beverage, and feed.
9. The product of claim 8, wherein the medicament further comprises a pharmaceutically acceptable excipient.
10. The product of claim 9, wherein the pharmaceutical is in a dosage form selected from any one of: granules, capsules, tablets, powders, oral liquids, suspensions, and emulsions.
11. The product according to claim 9, wherein the pharmaceutically acceptable excipient may be at least one selected from the group consisting of carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners, and flavors.
12. The product according to claim 9, wherein the pharmaceutically acceptable excipient is at least one selected from lactose, glucose, sucrose, sorbitol, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone (PVP), cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.
13. A Bacteroides culture method is characterized in that Bacteroides is inoculated on a slant under anaerobic condition and grows for 2-3 days in a culture medium on the slant at the temperature of 10-40 ℃; selecting thallus, inoculating the thallus into a culture solution, and culturing in the culture solution at 10-40 ℃ for 2-3 days, wherein the culture medium comprises 10-30g/L of carbon source, 6-15g/L of nitrogen source, 10-20g/L of agar and 0.2-1g/L of inorganic salt composition, and the pH value is 3.0-7.0; the culture solution comprises 10-30g/L of carbon source, 6-15g/L of nitrogen source, 1-3g/L of agar and 0.2-1g/L of inorganic salt composition, and the pH value is 3.0-7.0.
14. The method of claim 13, wherein the carbon source is fructose, glucose, or sucrose.
15. The method of claim 13, wherein the nitrogen source is corn steep liquor, yeast extract, or corn meal.
16. The method of claim 13, the inorganic salt composition comprising: potassium dihydrogen phosphate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, anhydrous calcium chloride and sodium chloride.
17. Use of bacteroides for the prevention and/or treatment of nicotine-related metabolic disorders.
18. The use according to claim 17, wherein the bacteroides is one or more of: bacteroides xylanolyticus, bacteroides dorsalis, bacteroides ovatus, bacteroides fragilis, bacteroides vulgatus.
19. A method of treating or preventing obesity-related disorders by providing a product according to any one of claims 5 to 12 to a target subject.
CN202211202491.9A 2022-09-29 2022-09-29 Application of bacteroides in preparation of product for preventing or treating nicotine-related metabolic diseases, related product and culture method Pending CN115400153A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106852938A (en) * 2015-12-09 2017-06-16 深圳华大基因研究院 Application of the bacteroid (Bacteroides) in obesity-related disease is treated and prevented
CN107002022A (en) * 2014-09-30 2017-08-01 上海交通大学医学院附属瑞金医院 Purposes of the bacteroid in obesity-related disease is treated or prevented
US20170224744A1 (en) * 2014-09-30 2017-08-10 Ruijin Hospital Affiliated To Shanghai Jiao Tong University School Of Medicine Uses of bacteroides in treatment or prevention of obesity and obesity-related diseases
CN107206030A (en) * 2014-12-08 2017-09-26 财团法人峨山社会福祉财团 Include the pharmaceutical compositions that are used to prevent or treat metabolic disease of the raw sour bacteroid as active ingredient
CN111714522A (en) * 2019-03-04 2020-09-29 中国科学院微生物研究所 Bacteroides and application thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107002022A (en) * 2014-09-30 2017-08-01 上海交通大学医学院附属瑞金医院 Purposes of the bacteroid in obesity-related disease is treated or prevented
US20170224744A1 (en) * 2014-09-30 2017-08-10 Ruijin Hospital Affiliated To Shanghai Jiao Tong University School Of Medicine Uses of bacteroides in treatment or prevention of obesity and obesity-related diseases
US20170252381A1 (en) * 2014-09-30 2017-09-07 Ruijin Hospital Affiliated To Shanghai Jiao Tong University School Of Medicine Uses of bacteroides in treatment or prevention of obesity and obesity-related diseases
CN107206030A (en) * 2014-12-08 2017-09-26 财团法人峨山社会福祉财团 Include the pharmaceutical compositions that are used to prevent or treat metabolic disease of the raw sour bacteroid as active ingredient
CN106852938A (en) * 2015-12-09 2017-06-16 深圳华大基因研究院 Application of the bacteroid (Bacteroides) in obesity-related disease is treated and prevented
CN111714522A (en) * 2019-03-04 2020-09-29 中国科学院微生物研究所 Bacteroides and application thereof

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