CN106974940A - Application of the heavy wall mushroom probiotics in fat and its relevant disease is treated and prevented - Google Patents

Application of the heavy wall mushroom probiotics in fat and its relevant disease is treated and prevented Download PDF

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Publication number
CN106974940A
CN106974940A CN201610032106.9A CN201610032106A CN106974940A CN 106974940 A CN106974940 A CN 106974940A CN 201610032106 A CN201610032106 A CN 201610032106A CN 106974940 A CN106974940 A CN 106974940A
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composition
bacterium
heavy wall
probiotics
fat
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CN106974940B (en
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冯强
张东亚
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BGI Shenzhen Co Ltd
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BGI Shenzhen Co Ltd
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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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • 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

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Nutrition Science (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses a kind of application of heavy wall mushroom probiotics in obesity-related disease is treated and prevented, specifically, heavy wall mushroom probiotics of the invention is used to prepare composition or preparation, and the composition or preparation are used for the one or more purposes being selected from the group：(a) prevent and/or treat fat；(b) blood fat is reduced；(c) prevent or treat angiocardiopathy；And/or (d) prevention and/or treatment diabetes.The heavy wall mushroom probiotics of the present invention can significantly reduce body weight, reduction blood fat, reduction body fat ratio.

Description

Application of the heavy wall mushroom probiotics in fat and its relevant disease is treated and prevented
Technical field
The invention belongs to microbiological art, in particular it relates to which heavy wall mushroom probiotics is treating and preventing fertilizer Application in fat and its relevant disease, is directed to the composition comprising heavy wall mushroom probiotics and its application.
Background technology
There are a large amount of symbiotic microorganisms in human body, they are largely resided in the enteron aisle of people, and quantity is more than 1,000,000,000,000,000 (1014The order of magnitude), it is more than 10 times of human body cell sum.In very long evolutionary process, enteric microorganism is reached with the mankind Good cooperation, nutrition, metabolism to human body and it is immune all play vital effect, many researchers be more by Human body intestinal canal microbiologic population regards one " organ " of human body, or the genome of human body second as, wherein the magnanimity contained Hereditary information is closely related with human health.By to nearly ten thousand kinds of hundreds of diseases such as diabetes, coronary heart disease, obesity, colon cancer The research of sample finds that some specific species show significant association with disease, clinical evaluation of these results in disease A brand-new direction is provided with the therapeutic intervention in diagnosis and later stage.
Obesity is a kind of chronic disease, and many factors can all cause obesity, and its origin of falling ill does not study clear so far.Fertilizer It is fat while being also a series of inducible factor of diseases, in such as hypertension, diabetes, coronary heart disease, cholecystopathy, osteoarthritis, sleep Breathing asphyxia, breathing imbalance, hysteroma, prostate cancer, breast cancer and colon cancer etc..According to NIH report, 9700 are there are about at present Ten thousand Americans are overweight and obesity, wherein the type ii diabetes number related to obesity reaches about 15,100,000 people, every year It there are about 200,000 people and die from the disease related to obesity.
Body fat is superfluous caused by obesity is often as physiology or biochemical function change.During fat is generally included Property fat, phosphatide and cholesterol.The increase of fat is due to the consumption that Energy intaking is more than energy.It is fat from nosogenesis There are two types：(a) simple obesity (simple obesity) and (b) Secondary Obesity (second obesity).Merely Property obesity can be divided into congenital obesity (idiopamic obesity) and posteriority fat (acquired obesity), it is single Pure property obese patient quantity can account for more than 95% total fat number.Congenital obesity be caused by substantial amounts of fat cell, and It is common in obesity in childhood.Posteriority obesity is that caused by larger sized fat cell, and it is fat to be common in the manhood.It is secondary Property obesity be otherwise known as symptomatic obesity, it is typically caused by endocrine or disease of metabolism.
There are five kinds of strategies for treating obesity at present：Go on a diet, take exercise, behaviour therapy, drug therapy and rehabilitation operation (therapetltic operation).Take the speed of the main health risk factor and weight loss for regarding patient of which kind of strategy Depending on effect, it may be selected or combine these strategies and adiposis patient is treated.The speed and effect of its weight loss are by all Such as influence of age, height, family history and risk factor Multiple factors.Diet-exercise regimen, that is, eat low in calories, low fat Food and carry out exercising with oxygen, but this method be generally acknowledged that it is unsuccessful to ordinary populace, and need adhere to regularly for a long time； The effect got instant result can be reached by removing body fat operation, but there are many restrictions, and such as operation risk, grease removal effect is difficult to hold Long and spend prohibitively expensive etc..
Drug therapy is current main clinical treatment obesity and its method for obesity-related disease (such as diabetes).Medicine The mechanism for the treatment of includes appetite-suppressing, increase energy expenditure, stimulates mobile fat, the synthesis of reduction triglycerides and suppress fat and inhale Receive.Main medicine is at present：Phenylpropanolamine (phenylpropanolamine, PPA), Xenical (orlistat, Xenical III) with Reductil (sibutramine, ReductilTM).The hyperglycaemia of some diabetes patients passes through diet and/or exercise regimen Or still can not obtain suitable control using above-mentioned therapeutic compounds.For these patients, exogenous insulin should be used.To patient For, the use of exogenous insulin is costly and painful method, can also bring multiple complications to patient.For example, due to not having Have and have a meal or abnormal exercise, the calculating mistake of insulin dose can cause insulin response (hypoglycemia).In addition, using medicine It may also happen that to the locally or systemically allergy or immune resistance of medicine.
A kind of method of current this area also treatment and prevention obesity and its relevant disease without effective, Small side effects And medicine.
Therefore this area is a kind of new in the urgent need to developing, and has no toxic side effect, for treat and prevent it is fat and its The medicine of relevant disease.
The content of the invention
It is an object of the present invention to provide heavy wall mushroom probiotics in terms of fat and its relevant disease is treated and prevented Purposes.
Effectively had no toxic side effect it is a further object of the present invention to provide a kind of, for treating and preventing fat and its phase Medicine, beverage, the food compositions of related disorders, or animal feed composition.
It is a further object of the present invention to provide a kind of losing weight and/or the method and its application of blood glucose.
First aspect present invention provides a kind of purposes of heavy wall mushroom probiotics, for preparing composition or preparation, institute State one or more purposes that composition or preparation are used to be selected from the group：(a) prevent and/or treat fat；(b) blood fat is reduced； (c) prevent or treat angiocardiopathy；And/or (d) prevention and/or treatment diabetes, wherein, the heavy wall mushroom probiotics choosing From the following group：Association fecal bacteria (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister invisus), Or its combination.
In another preference, the heavy wall mushroom probiotics includes association fecal bacteria (Coprococcus eutactus)。
In another preference, the association fecal bacteria (Coprococcus eutactus) is selected from the group： Coprococcus eutactus ATCC 27759、Coprococcus eutactus ATCC 51897、Coprococcus Eutactus L2-50 or its combination.
In another preference, the heavy wall mushroom probiotics wears Alister bacterium (Dialister including muddiness invisus)。
In another preference, the muddiness is worn Alister bacterium (Dialister invisus) and is selected from the group： Dialister invisus DSM 15470、Dialister invisus E2.20、Dialister invisus E9.48、 Or its combination.
In another preference, the heavy wall mushroom probiotics includes the one or more in table 3.
In another preference, the heavy wall mushroom probiotics is selected from table 3, and from identical or different category.
In another preference, the composition is selected from the group：Food compositions, health composition, pharmaceutical composition, drink Feed composition, fodder compound or its combination.
In another preference, described composition is oral formulations.
In another preference, described composition is liquid formulation, solid formulation, semisolid preparations.
In another preference, the formulation of described composition is selected from the group：Powder agent, powder, tablet, sugar-coat agent, glue Wafer, granule, suspending agent, solution, syrup, drops and sublingual lozenge.
In another preference, described food compositions include latex product, solution product, pulverulent product or suspended Liquid product.
In another preference, described food compositions include dairy products, milk powder or emulsion.
In another preference, described liquid formulation is selected from the group：Solution product or suspension product.
Second aspect of the present invention provides a kind of purposes of heavy wall mushroom probiotics, for preparing composition or preparation, institute State one or more purposes that composition or preparation are used to be selected from the group：(i) reduction mammal in MCP- 1 (MCP-1) level；And/or (ii) improves leptin resistance, the sensitiveness in vivo to Leptin is improved, wherein, the firmicutes Class probiotics is selected from the group：Association fecal bacteria (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister Invisus) or its combination.
In another preference, the composition or preparation are also independently or additionally used for the one kind or many being selected from the group Plant purposes：
(iii) body weight increase of mammal is suppressed；
(iv) the body fat ratio (the ratio between fat weight/body weight) of mammal is reduced；
(v) blood lipid level of mammal is reduced；
(vi) level of the HDL (HDLC) in mammal is improved；
(vii) low-density lipoprotein (LDLC) level in reduction mammal.
In another preference, the mammal includes people, rodent (such as rat, mouse).
In another preference, it is described reduction mammal blood lipid level include reduction T-CHOL (TC) level and/ Or triglyceride levels.
Third aspect present invention is used for the composition for the treatment of and/or pre- preventing obesity there is provided a kind of, and the composition includes： (i) the heavy wall mushroom probiotics of safe and effective amount；On (ii) food or pharmaceutically acceptable carrier；Wherein, the heavy wall Mushroom probiotics is selected from the group：Association fecal bacteria (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister invisus) or its combination.
In another preference, the heavy wall mushroom probiotics includes association fecal bacteria (Coprococcus eutactus)。
In another preference, the association fecal bacteria (Coprococcus eutactus) is selected from the group： Coprococcus eutactus ATCC 27759、Coprococcus eutactus ATCC 51897、Coprococcus Eutactus L2-50 or its combination.
In another preference, the heavy wall mushroom probiotics wears Alister bacterium (Dialister including muddiness invisus)。
In another preference, the muddiness is worn Alister bacterium (Dialister invisus) and is selected from the group： Dialister invisus DSM 15470、Dialister invisus E2.20、Dialister invisus E9.48、 Or its combination.
In another preference, the composition is selected from the group：Food compositions, health composition, pharmaceutical composition, drink Feed composition, fodder compound or its combination.
In another preference, the composition contains 1 × 10-1 × 1020Cfu/mL or cfu/g heavy wall mushroom is prebiotic Bacterium, preferably 1 × 104-1×1015Cfu/mL or cfu/g heavy wall mushroom probiotics, by the cumulative volume or total of the composition Weight meter.
In another preference, in described composition, containing 0.0001-99wt%, preferably described in 0.1-90wt% Heavy wall mushroom probiotics, with the gross weight meter of the composition.
In another preference, described composition is unit dosage form (an a piece of, capsule or a bottle), each unit The quality of composition described in formulation is 0.05-5g, preferably 0.1-1g.
In another preference, described composition also contains other probiotics and/or prebiotics.
In another preference, other described probiotics are selected from the group：Lactic acid bacteria, Bifidobacterium, lactobacillus acidophilus or It is combined.
In another preference, described prebiotics is selected from the group：It is FOS (FOS), galactooligosaccharide (GOS), low Xylan (XOS), lactosucrose (LACT), soyabean oligosaccharides (SOS), inulin (Inulin) or its combination.
Fourth aspect present invention provides a kind of preparation method of composition described in third aspect present invention, including step：
Mixed by (i) heavy wall mushroom probiotics, with acceptable carrier or pharmaceutically acceptable carrier on (ii) food, So as to form the composition described in third aspect present invention.
In another preference, described composition is oral formulations.
A kind of method that fifth aspect present invention provides losing weight and/or blood fat, (i) heavy wall is applied to the object Composition described in mushroom probiotics or third aspect present invention.
In another preference, described administration includes oral.
In another preference, described application dosage is 0.01-5g/50kg body weight/days, it is preferred that 0.1-2g/50kg Body weight/day.
In another preference, described object includes mammal, such as people.
It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the invention and have in below (eg embodiment) It can be combined with each other between each technical characteristic of body description, so as to constitute new or preferred technical scheme.As space is limited, exist This no longer tires out one by one states.
Brief description of the drawings
Fig. 1 shows body weight increase situation of each group mouse compared with before gavage after gavage association fecal bacteria.
Fig. 2 shows that gavage muddiness wears body weight increase situation of each group mouse compared with before gavage after Alister bacterium.
Fig. 3 shows body weight increase situation of each group mouse compared with before gavage after gavage combination strain.
Fig. 4 shows the body fat ratio of gavage association fecal bacteria each group mouse after 9 weeks.
Fig. 5 shows that gavage muddiness wears the body fat ratio of Alister bacterium each group mouse after 9 weeks.
Fig. 6 shows the body fat ratio of gavage combination strain each group mouse after 9 weeks.
Fig. 7 shows influence of the gavage association fecal bacteria to blood fat.
Fig. 8 shows that gavage muddiness wears influence of the Alister bacterium to blood fat.
Fig. 9 shows influence of the gavage combination strain to blood fat.
Figure 10 shows influence of the gavage association fecal bacteria to monocyte chemoattractant protein-1 (MCP-1).
Figure 11 shows that gavage muddiness wears influence of the Alister bacterium to monocyte chemoattractant protein-1 (MCP-1).
Figure 12 shows influence of the gavage combination strain to monocyte chemoattractant protein-1 (MCP-1).
Figure 13 shows influence of the gavage association fecal bacteria to leptin (Leptin, LEP).
Figure 14 shows that gavage muddiness wears influence of the Alister bacterium to leptin (Leptin, LEP).
Figure 15 shows influence of the gavage combination strain to leptin (Leptin, LEP).
Embodiment
The present inventor is by in-depth study and experiment extensively, it has unexpectedly been found that, association fecal bacteria (Coprococcus ) and/or muddiness wears Alister bacterium (Dialister invisus) and has prevention and treatment fat and its related disease eutactus The effect of sick (such as angiocardiopathy), by the active compound feeding food experimental subjects containing above-mentioned heavy wall mushroom probiotics, hair Existing said composition can suppress increased weight, reduce body fat ratio, reduce blood fat, effectively mitigate the illnesss such as cardiovascular and obesity. The present invention is completed on the basis of this.
As used herein, term " containing " represents that various composition can be applied to the mixture or composition of the present invention together In.Therefore, term " mainly by ... constitute " and " consist of " are included in term " containing ".
As used herein, described " body fat ratio " refers to the ratio of fat weight/body weight.
The heavy wall mushroom probiotics of the present invention and its application
As used herein, a class Firmicutes of commensalism in described " heavy wall mushroom probiotics of the invention " duodenum 12 road Bacterium, Gram's staining is positive, shaft-like, such as clostridium XIVa clusters and IV clusters (Clostridium cluster XIVa and IV), Including fusobacterium (Clostridium), Eubacterium (Eubacterium), Ruminococcus (Ruminococcus) and sulfuric acid Some kinds of salt reduction Pseudomonas (Anaerofilum) etc..By with other microbial interactions of enteron aisle, such probiotics is in intestines Played a significant role in road colony balance, meanwhile, also play other specific or required functions.
In the present invention, described " heavy wall mushroom probiotics of the invention " refers to association fecal bacteria (Coprococcus Eutactus), muddiness wear one or more Pseudomonas of Alister bacterium (Dialister invisus) 2 kinds of Pseudomonas mixing or One or more mixing of multiple bacterium in each Pseudomonas.
Wherein, association fecal bacteria (Coprococcus eutactus) is a kind of obligate anaerobe, it is impossible to moves, works as presentation Gram's staining is positive when going out paired or chain pair state, but can fade quickly.Containing fermentable carbohydrate In culture medium, slightly elongated shape is presented.But the bacterium is generally circular, a diameter of 0.7-1.3 μm.
It is a kind of obligate anaerobe, not movable coccobacillus that muddiness, which wears Alister bacterium (Dialister invisus), (0.3-0.4 × 0.3-0.6 μm), Gram's staining is negative.Single dispersing morphology can be presented, can also be rendered into, short chain, small The forms such as cluster.
Fat and its relevant disease (such as angiocardiopathy) is being treated and prevented the invention provides heavy wall mushroom probiotics The application of aspect.Subject takes in high-fat food, and there is heavy wall mushroom probiotics (i) to suppress subject's increased weight； (ii) blood fat is reduced；(iii) reduce body fat than ability.According to the preference of the present invention, the firmicutes through the present invention Class probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can lead obesogenous high fat being fed for treating The C57BL/6J male mices of the food of fat, compared with not receiving the control group for the treatment of, its increased weight amplitude slows down and blood fat Decline, and it is various also decline to fat or related angiocardiopathy index, such as leptin (LEP) and monocyte chemotactic egg - 1 (MCP-1) in vain.Therefore, heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its group Close) can be to prevent and treat the fat and disease as caused by obesity, such as angiocardiopathy.
Composition and its application
Present invention also offers a kind of composition, it is preferable that is pharmaceutical composition.The composition includes the sheet of effective dose The heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) of invention, in a preference, institute State the probiotics that composition also includes being selected from the group：Lactic acid bacteria, Bifidobacterium, lactobacillus acidophilus or its combination；And/or be selected from The prebiotics of the following group：It is FOS (FOS), galactooligosaccharide (GOS), xylo-oligosaccharide (XOS), lactosucrose (LACT), big Beans oligosaccharide (SOS), inulin (Inulin) or its combination.
In a preference, described composition is liquid formulation, solid formulation, semisolid preparations.
In a preference, described liquid formulation is selected from the group：Solution product or suspension product.
In a preference, the formulation of described composition is selected from the group：Powder agent, powder, tablet, sugar-coat agent, capsule Agent, granule, suspending agent, solution, syrup, drops and sublingual lozenge.
Pharmaceutical composition of the present invention can be with medicinal tablet, any form administration of injection or capsule, the medicine system Agent includes the medium and carrier that excipient, medicine allow, and these materials can be selected according to method of administration.Medicine in the present invention Preparation can further include the active constituent of auxiliary.
Lactose, glucose, sucrose, D-sorbite, mannose, starch, Arabic gum, calcium phosphate, alginates, gelatin, silicon Sour calcium, fine crystallization cellulose, polyvinylpyrrolidone (PVP), cellulose, water, syrup, methylcellulose, hydroxybenzoic acid first Ester, nipasol, talcum, magnesium stearate or mineral oil etc. are used as the carrier of pharmaceutical composition in the present invention, tax Shape agent or diluent etc..
In addition, the present invention pharmaceutical composition can further comprise lubricant, wetting agent, emulsifying agent, suspension stabiliser, Preservative, sweetener and spices etc..The pharmaceutical composition of the present invention can be produced by a variety of known methods with enteric-coated preparations, with Stomach can be passed through without being destroyed by hydrochloric acid in gastric juice by being easy to the active component i.e. microorganism of pharmaceutical composition.
In addition, the capsule form that the microorganism of the present invention can prepare in conventional manner is used.For example, standard excipients and sheet The cold dry microorganism of invention is mixed and made into bead pill, and then pill is packed into gelatine capsule.In addition, micro- life of the present invention Thing and medicine allow the excipient used such as liquid glue, cellulose, silicate or mineral oil etc. suspension to be made by mixing or scattered Liquid, this suspension or dispersion liquid can be fitted into soft gelatine capsule.
The pharmaceutical composition of the present invention can be made into casing piece for being administered orally.Term-" casing " in the application, including All conventional medicines allow the coating used, and these are coated not by gastric acid degradation, but can fully be decomposed and fast quick-release in small intestine Release the microorganism of the present invention.The casing of wood invention can be small in 36-38 DEG C of maintenance 2 in synthesis hydrochloric acid in gastric juice such as pH=1 HCl solution When more than, and preferably synthesis intestinal juice such as pH=7.0 buffer solution in 1.0 hours decomposition.
The casing of the present invention is to be coated with every agreement that contracts a film or TV play to an actor or actress 16-30mg, preferably 16-25mg, and more preferably 16-20mg is carried out It is coated.Casing thickness is 5-100 μm in the present invention, and preferable thickness is 20-80 μm.Casing composition select oneself it is open know it is normal Advise polymer.
Currently preferred casing by cellulosic phthalic acetate polymer or trimellitic acid ester polymer and Methacrylate copolymer (for example, containing more than 40% methacrylate and containing methyl cellulose phthalate hydroxypropyl acrylate or The copolymer of the methacrylate of its ester derivative) prepare.
The viscosity of cellulosic phthalic acetate used in casing is about 45-90cp, acetyl content in the present invention 17-26%, phthalate content 30-40%.It is about 5-21cp for the inclined ester of phthalic acid viscosity of cellulose acetate in casing, Second phthalein content 17-26%.Cellulose acetate trimellitate is produced by Eastman Kodaks company, the intestines that can be used in the present invention Clothing material.
Cruel for the hydroxypropyl methyl cellulose phthalic acid in casing of the present invention, molecular weight is generally 20,000- 130,000 dalton, desired molecular weight is 80,000-100,000 dalton, and hydroxypropyl content is 5-10%, methoxyl content For 18-24%, phthalyl content is 21-35%.
It is extremely HP50 for the hydroxypropyl methyl cellulose phthalic acid in casing of the present invention, by Japanese Shin-Etsu Chemidnl Co.Ltd. are produced.HP50 contains 6-10% hydroxypropyls, 20-24% methoxyl groups, 21-27% propyl group, its molecule Measure as 84,000 dalton.Another Enteric materials are HP55, and HP55 contains 5-9% hydroxypropyl methyl cellulose O-phthalic Acid esters, 18-22% methoxyl groups, 27-35% phthalic acid, its molecular weight is 78,000 dalton.
Casing of the present invention is prepared as follows：Casing solution is sprayed in core using conventional method.In the enteric coating method All solvents are alcohols (such as ethanol), ketone (such as acetone), halogenated hydrocarbon compound (such as dichloromethane) or its composition.Will be soft Agent such as di-n-butyl phthalic acid ester and glyceryl triacetate are added in casing solution, and its ratio is 1 part of coatings pair About 0.05 part or about 0.3 part of softening agent.Spray method is preferably continuously performed, and the doses sprayed can be according to the used bar of coating Part is controlled.Atomisation pressure can be adjusted arbitrarily, it is however generally that, preferable result can be obtained under average 1-1.5 handkerchiefs pressure.
In specification " medicine effective quantity " refer to that people and/or animal can be produced function or activity and can by people and/or The amount that animal is received.Such as, in the present invention, it can prepare containing 1 × 10-1 × 1020Cfu/ml or cfu/g (particularly, can Contain 1 × 104-1×1015Cfu/ml or cfu/g；More particularly, 1 × 10 can be contained6-1×1011Cfu/ml or cfu/g) sheet The preparation of the heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) of invention.
When for preparing pharmaceutical composition, heavy wall mushroom probiotics (such as association fecal bacteria, muddiness of the invention used Wear Alister bacterium or its combination) effective dose can change with the pattern of administration and the order of severity of disease to be treated. Suitable for dosage form for oral administration, comprising with solid-state or the intimately mixed about 1 × 10-1 of liquid pharmaceutically acceptable carrier × 1020Cfu/ml or cfu/g (particularly, can contain 1 × 104-1×1015Cfu/ml or cfu/g；More particularly, can containing 1 × 106-1×1011Cfu/ml or cfu/g) active heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or Its combine) active component.This dosage is can adjust to provide optimal treatment response.For example, by treatment situation it is urgent highly necessary Ask, dosage separated several times can be given daily, or dosage is reduced pari passu.
Described heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can be by oral Given etc. approach.Solid-state carrier includes：Starch, lactose, Dicalcium Phosphate, microcrystalline cellulose, sucrose and white bole, and liquid carrier Including：Culture medium, polyethylene glycol, nonionic surface active agent and edible oil (such as corn oil, peanut oil and sesame oil), as long as It is adapted to heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) characteristic and required specific administration Mode.Usually used adjuvant also can be advantageously included in pharmaceutical composition is prepared, such as flavor enhancement, pigment, preservative With antioxidant such as vitamin E, vitamin C, BHT and BHA.
In terms of easily prepared and administration position, pharmaceutical composition preferably is solid-state composition, especially tablet and solid The capsule that body is filled or liquid is filled.Oral administration is preferred.
The present composition is administered to the individual, is administered once daily or repeatedly.Dosage unit represents its form On can separate and suitable for the dosage of the mankind or other all mammalian subjects.Per unit contain medicine permission carrier and The microorganism of the present invention of effective therapeutic dose.Dosage with patient body weight and the fat order of severity, included supplement activearm Part and used microorganism and change.In addition as may, can separate and be administered, and if desired for can successive administration.Therefore, it is described Dosage will not cause limitation to the present invention.In addition, " composition " in the present invention does not mean only that medicine and expression can be made For functional food and healthy supplement.In a preference, the composition includes：Beverage, food, medicine, animal Feed etc..
In the preference of the present invention, a kind of food compositions are additionally provided, it contains the heavy wall mushroom of effective dose Probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its combination), and acceptable carrier on the food of surplus, institute The formulation for the food composition stated is selected from solid, dairy products, solution product, pulverulent product or suspension product.
In a preference, the formula of the composition is as follows：
1×10-1×1020Cfu/mL heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its Combination)；And on food or pharmaceutically acceptable carrier, and/or excipient.
In another preference, the formula of the composition is as follows：
1×106-1×1011Cfu/mL heavy wall mushroom probiotics (such as association fecal bacteria, muddiness wear Alister bacterium or its Combination)；And on food or pharmaceutically acceptable carrier, and/or excipient.
Losing weight and/or the method for blood fat
In another preference, methods described includes：Absorb pharmaceutical composition, food compositions, the beverage group of the present invention Compound or its combination.The experimental subjects is behaved.
In another preference, methods described includes：Absorb pharmaceutical composition, food compositions or the animal of the present invention Feed, or its combination.The experimental subjects is animal, preferably muroid, Lagomorpha.
Main advantages of the present invention include：
(a) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Significantly reduce body weight, reduction blood fat, reduction body fat ratio.
(b) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Significantly reduce and fat and its related relevant disease (such as angiocardiopathy) index (such as cholesterol and triglycerides).
(c) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Significantly reduce the level of T-CHOL, triglycerides, low-density lipoprotein.
(d) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Significantly improve the level of HDL.
(e) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Improve insulin resistance, can also reduce the risk that atherosclerosis and angiocardiopathy occur.
(f) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Significantly reduce monocyte chemoattractant protein-1 (MCP-1) level.
(g) heavy wall mushroom probiotics of the invention (such as association fecal bacteria, muddiness wear Alister bacterium or its combination) can Effectively improve the adjoint leptin resistance of obesity, improve the sensitiveness in vivo to Leptin.
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention Rather than limitation the scope of the present invention.The experimental method of unreceipted actual conditions in the following example, generally according to conventional strip Part such as Sambrook et al., molecular cloning：Laboratory manual (New York：Cold Spring Harbor Laboratory Press, 1989) described in condition, or according to《Microorganism：Laboratory manual》(James Cappuccino and Natalie Sherman is compiled, Pearson Education publishing houses) described in condition, or according to the condition proposed by manufacturer.
The food compositions of the component containing bacterium of embodiment 1 (such as association fecal bacteria, muddiness wear Alister bacterium or its combination)
Raw material proportioning such as table 1.
The Combined food composition formula of table 1
Raw material Mass percent (%)
Bacterium component 0.5
Milk 90.0
White sugar 9.5
The bacterium component being formulated in 1-6 is single bacteria component, respectively containing Coprococcus eutactus ATCC 27759、Coprococcus eutactus ATCC 51897、Coprococcus eutactus L2-50、Dialister invisus DSM 15470、Dialister invisus E2.20、Dialister invisus E9.48。
Bacterium component in formula 7 is the mixture of any two or more (preferably 2 kinds or 3 kinds) of above-mentioned 6 kinds of bacterium (weight ratio is 1:1 or 1:1:1).
According to above-mentioned formula rate mixing milk, white sugar, stirring is preheated, 20Mpa pressure homogeneous, 90 DEG C to being thoroughly mixed Left and right is sterilized 5-10 minutes, is cooled to 40-43 DEG C, is inoculated with 1-100 × 106Cfu/g bacterium component, that is, be made component containing bacterium (such as Association fecal bacteria, muddiness wear Alister bacterium or its combination) food compositions.
Embodiment 2
The pharmaceutical composition of component containing bacterium (such as association fecal bacteria, muddiness wear Alister bacterium or its combination)
Raw material proportioning is shown in Table 2.
The drug regimen composition formula of table 2
Raw material Mass percent (%)
Bacterium component 1.0%
Lactose 2.0%
Dusty yeast 2.0%
Peptone 1.0%
Pure water 94.0%
The bacterium component being formulated in 1-6 is single bacteria component, respectively containing Coprococcus eutactus ATCC 27759、Coprococcus eutactus ATCC 51897、Coprococcus eutactus L2-50、Dialister invisus DSM 15470、Dialister invisus E2.20、Dialister invisus E9.48。
Bacterium component in formula 7 is the mixture of any two or more (preferably 2 kinds or 3 kinds) of above-mentioned 6 kinds of bacterium (weight ratio is 1:1 or 1:1:1).Proportionally lactose, dusty yeast, peptone are well mixed with pure water, 60-65 is preheating to DEG C, 20Mpa pressure homogeneous, 90 DEG C or so are sterilized 20-30 minutes, are cooled to 36-38 DEG C, access bacterium component (1-50 × 106cfu/ ML), 36-38 DEG C of fermentation to pH value is 6.0, and centrifugation, freeze-drying to water content is less than 3%, that is, is prepared into the component containing bacterium It is freeze-dried thing.The freeze-drying thing of 0.5 gram of component containing bacterium is weighed with being fitted into after maltodextrin mixed in equal amounts in capsule, that is, is made The pharmaceutical composition of component containing bacterium (such as association fecal bacteria, muddiness wear Alister bacterium or its combination).
Therapeutic action of the embodiment 3 for obese model mouse
Experiment material:
Mouse：C57BL/6J male mices (being purchased from Guangdong Medical Lab Animal Center) are bought, are normal raising mouse, 6 week old.Mouse growth process is in same environment, and feeds same food.
6 plants of heavy wall mushroom probiotics are obtained from preservation mechanism, and are stored in BGI-Shenzhen.Meanwhile, choose Lactobacillus plantarum (Lactobacillus plantarum), from the common micro- life of China Committee for Culture Collection of Microorganisms Thing center (CGMCC), deposit number CGMCC No.8198, as a control group (LP groups), and in MRS nutrient solutions, 37 DEG C of cultures 24-48h。
Wherein, the source-information of above-mentioned 6 plants of heavy walls mushroom probiotics is as shown in table 3.All bacterial strains are sequenced through 16S rDNA Start experiment after identification is errorless.
Bacterial strain information is shown in Table 3.
The bacterial strain information of table 3
Wherein, Coprococcus eutactus L2-50 (bacterium 3) derive from Britain sieve Witter research institute (Rowett Research Institute)(Barcenilla A,Pryde SE,Martin JC et al.Phylogenetic Relationships of Butyrate-Producing Bacteria from the Human Gut.Applied and Environmental Microbiology 2000；66:1654-1661.).Dialister invisus E2.20 (bacterium 5) and Dialister invisus E9.48 (bacterium 6) derive from dentistry research institute of Gai Shi hospitals of London King's College microorganism Section (Department of Microbiology, Dental Institute, Guy's Tower, Guy's Hospital, King's College London,UK；J.Downes, M.Munson and W.G.Wade, Dialister invisus sp.nov.,isolated from the human oral cavity,International Journal of Systematic and Evolutionary Microbiology(2003),53,1937–1940)。
High lipid food (HF)：Containing 78.8% basal feed, 1% cholesterol, 10% yolk powder, 10% lard and 0.2% courage Salt, purchased from Nantong Te Luofei feed technologies Co., Ltd.
It is common to maintain feed：Purchased from Guangdong Medical Lab Animal Center.
Experimental method：
Choose the C57BL/6J adult male mices normally fed, respectively random packet, control group (CK), microbial inoculum group (1 group of bacterium, 2 groups of bacterium, 3 groups of bacterium, 4 groups of bacterium, 5 groups of bacterium, 6 groups of bacterium, 5 groups of bacterium 1+ bacterium, 6 groups of bacterium 2+ bacterium, 4 groups of bacterium 3+ bacterium), compares microbial inoculum Group (LP, Lactobacillus plantarum CGMCC No.8198) and obese model group (HF), every group 10, in SPF Ad lib and drinking-water under (no-special pathogen (Specific pathogen Free)) environment.LP groups, HF groups and microbial inoculum group Feeding high lipid food, CK group feedings commonly maintain feed.Feed after 4 weeks, microbial inoculum group and LP groups start the corresponding bacterial strain bacterium solution of gavage； HF groups and CK group gavage equivalent culture mediums, gavage 9 weeks.
Feed bacterium amount and be set to 0.15mL/10g body weight, dense bacterium is 1 × 107Concentration is 1 × 10 after cfu/mL, concentration8cfu/ ML, frequency is once every other day.Bacterium solution need to be cultivated in advance, and activation weekly ensures fresh, determines concentration respectively, and adjust to 1 × 108cfu/mL.For single microbial inoculum group, corresponding bacterium solution is taken by above-mentioned dosage gavage；For mix bacterium agent group, by each single bacterium bacterium solution Above-mentioned dosage gavage is pressed after equal proportion mixing.
In experiment periods, the data such as mouse weight, state, food-intake are recorded weekly.Test each group mouse progress in last week Glucose tolerance (OGTT) is tested.Mouse is put to death in experiment after terminating, record fat weight, and takes serum, is examined with Elisa kits Survey blood fat and protein factor content.
Experimental result：
(1) association fecal bacteria, muddiness wear the influence of Alister bacterium or its combination strain to mouse body weight.
Body weight increase situation (Fig. 1) of each group mouse compared with before gavage after the gavage association fecal bacteria of table 4
Note：Data are mean+SD in table, for any two groups of data of each row, not identical behind numeral Letter represents significant difference (p<0.05), table 5-15 is identical with this.
The gavage muddiness of table 5 wears body weight increase situation (Fig. 2) of each group mouse compared with before gavage after Alister bacterium
Body weight increase situation (Fig. 3) of each group mouse compared with before gavage after the gavage combination strain of table 6
As a result as shown in table 4-6 and Fig. 1-3.As a result show, association fecal bacteria, muddiness wear Alister bacterium or its combination Growth (the P of the body weight of obese model mouse can effectively be slowed down<0.05).
(2) association fecal bacteria, it is muddy wear Alister bacterium or its combination strain to body fat than influence.
The body fat ratio (Fig. 4) of the gavage association fecal bacteria of table 7 each group mouse after 9 weeks
Packet Fat weight/body weight × 100%
CK 2.83±0.13e
Bacterium 1 4.11±0.10c
Bacterium 2 4.00±0.26cd
Bacterium 3 3.85±0.19d
LP 5.39±0.34b
HF 7.48±0.46a
The gavage muddiness of table 8 wears the body fat ratio (Fig. 5) of Alister bacterium each group mouse after 9 weeks
Packet Fat weight/body weight × 100%
CK 2.91±0.14e
Bacterium 4 3.97±0.22cd
Bacterium 5 4.16±0.23c
Bacterium 6 3.76±0.20d
LP 5.21±0.24b
HF 746±064a
The body fat ratio (Fig. 6) of 9 gavage combination strain of table each group mouse after 9 weeks
Packet Fat weight/body weight × 100%
CK 2.90±0.13d
Bacterium 1+5 3.73±0.19c
Bacterium 2+6 3.87±0.23c
Bacterium 3+4 3.93±0.22c
LP 5.42±0.17b
HF 756±055a
As a result as shown in table 7-9 and Fig. 4-6.As a result show, association fecal bacteria, muddiness wear Alister bacterium or its combination strain Body fat ratio (the P of obese model mouse can be significantly reduced<0.05).
(3) association fecal bacteria, muddiness wear the influence of Alister bacterium or its combination strain to blood fat.
The gavage association fecal bacteria of table 10 each group lipid of mice content (Fig. 7) after 9 weeks
Packet TC(mmol/L) TG(mmol/L) LDLC(mmol/L) HDLC(mmol/L)
CK 3.811±0.144d 0.914±0.048d 1.260±0.057e 3.354±0.166a
Bacterium 1 4.819±0.198c 1.041±0.080c 1.516±0.084c 3.529±0.123a
Bacterium 2 4.790±0.226c 1.052±0.059c 1.479±0.083cd 3.510±0.160a
Bacterium 3 4.823±0.251c 1.073±0.057c 1.457±0.102d 3.479±0.158a
LP 5.385±0.253b 1.234±0.094b 1.872±0.124b 2.743±0.139b
HF 6299±0257a 1303±0076a 2381±0157a 2193±0104c
The gavage muddiness of table 11 wears Alister bacterium each group lipid of mice content (Fig. 8) after 9 weeks
Packet TC(mmol/L) TG(mmol/L) LDLC(mmol/L) HDLC(mmol/L)
CK 3.892±0.154d 0.983±0.047c 1.250±0.036d 3.376±0.202a
Bacterium 4 4.896±0.230c 1.012±0.072c 1.395±0.082c 3.497±0.093a
Bacterium 5 4.906±0.247c 1.014±0.054c 1.353±0.076c 3.439±0.160a
Bacterium 6 4.917±0.201c 1.069±0.070c 1.364±0.071c 3.421±0.195a
LP 5.258±0.302b 1.216±0.088b 1.831±0.131b 2.741±0.100b
HF 6.390±0.290a 1.295±0.064a 2.357±0.126a 2.153±0.105c
12 gavage combination strain of table each group lipid of mice content (Fig. 9) after 9 weeks
Packet TC(mmol/L) TG(mmol/L) LDLC(mmol/L) HDLC(mmol/L)
CK 3.902±0.111d 0.964±0.046d 1.213±0.060d 3.287±0.239a
Bacterium 1+5 4.822±0.139c 1.019±0.067c 1.369±0.077c 3.324±0.177a
Bacterium 2+6 4.904±0.182c 1.017±0.061c 1.367±0.077c 3.249±0.215a
Bacterium 3+4 4.834±0.192c 1.009±0.060c 1.381±0.082c 3.252±0.140a
LP 5.498±0.280b 1.213±0.061b 1.742±0.105b 2.621±0.106b
HF 6.511±0.252a 1.304±0.094a 2.357±0.166a 2.196±0.096c
As a result as shown in Fig. 7-9 and table 10-12.Main Ingredients and Appearance in blood fat is courage in cholesterol and triglycerides, blood plasma The rise of sterol and triglyceride level is relevant with the generation of atherosclerosis.As a result show, association fecal bacteria, muddiness wear Ah Li Site bacterium or its combination strain can reduce blood fat, and the correlation of reduction atherosclerosis relevant disease (such as angiocardiopathy) refers to Mark.Also, association fecal bacteria, muddiness wear Alister bacterium or its combination strain reduction T-CHOL (TC), total triglycerides (TG) With low-density lipoprotein (LDLC), and increase HDL (HDLC) particularly evident (P of effect<0.05).
(4) association fecal bacteria, muddiness wear Alister bacterium or its combination strain leptin (Leptin, LEP), monocyte are become Change the influence of albumen -1 (MCP-1).
The gavage association fecal bacteria of table 13 each group mouse Leptin (Leptin, LEP), monocyte chemoattractant protein-1 after 9 weeks (MCP-1) content (Figure 10, Figure 13)
The gavage muddiness of table 14 wears Alister bacterium each group mouse Leptin (Leptin, LEP), monocyte chemotactic egg after 9 weeks - 1 (MCP-1) content (Figure 11, Figure 14) in vain
Packet MCP-1(pg/ml) LEP(pg/ml)
CK 343.07±44.17c 1162.93±96.41c
Bacterium 4 339.99±39.03c 1158.48±130.63c
Bacterium 5 338.79±31.89c 1166.22±136.26c
Bacterium 6 336.81±38.48c 1158.43±111.51c
LP 365.95±48.92b 1264.09±158.29b
HF 383.04±45.92a 1401.25±147.52a
15 gavage combination strain of table each group mouse Leptin (Leptin, LEP), monocyte chemoattractant protein-1 (MCP-1) after 9 weeks Content (Figure 12, Figure 15)
Packet MCP-1(pg/ml) LEP(pg/ml)
CK 336.55±38.88c 1164.35±106.96c
Bacterium 1+5 335.85±31.27c 1173.25±114.83c
Bacterium 2+6 333.48±31.21c 1169.25±132.61c
Bacterium 3+4 330.36±35.21c 1176.85±109.40c
LP 366.47±24.75b 1259.06±144.03b
HF 384.12±44.29a 1401.18±179.66a
As a result as shown in Figure 10-15 and table 13-15.As a result show, association fecal bacteria, muddiness wear Alister bacterium or its group The content of leptin (LEP) and monocyte chemoattractant protein-1 (MCP-1) in obese model mice serum can substantially be reduced by closing bacterium (P<0.05)。
As a result show, association fecal bacteria, muddiness wear Alister bacterium or its combination strain can improve leptin resistance, improve body The interior sensitiveness to leptin (LEP)；Also, the blood after association fecal bacteria, muddiness wear Alister bacterium or the treatment of its combination strain Clear MCP-1 levels reduction, is conducive to improving insulin resistance, it is possible to decrease the wind that atherosclerosis and angiocardiopathy occur Danger.
All documents referred in the present invention are all incorporated as reference in this application, independent just as each document It is incorporated as with reference to such.In addition, it is to be understood that after the above-mentioned instruction content of the present invention has been read, those skilled in the art can To be made various changes or modifications to the present invention, these equivalent form of values equally fall within the model that the application appended claims are limited Enclose.

Claims (10)

1. a kind of purposes of heavy wall mushroom probiotics, it is characterised in that for preparing composition or preparation, the composition or system Agent is used for the one or more purposes being selected from the group：(a) prevent and/or treat fat；(b) blood fat is reduced；(c) prevent or treat Angiocardiopathy；And/or (d) prevention and/or treatment diabetes, wherein, the heavy wall mushroom probiotics is selected from the group：Association excrement Coccus (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister invisus) or its combination.
2. purposes as claimed in claim 1, it is characterised in that the heavy wall mushroom probiotics includes association fecal bacteria (Coprococcus eutactus)。
3. purposes as claimed in claim 2, it is characterised in that association fecal bacteria (Coprococcus eutactus) choosing From the following group：Coprococcus eutactus ATCC 27759、Coprococcus eutactus ATCC 51897、 Coprococcus eutactus L2-50 or its combination.
4. a kind of purposes of heavy wall mushroom probiotics, it is characterised in that for preparing composition or preparation, the composition or system Agent is used for the one or more purposes being selected from the group：(i) water of monocyte chemoattractant protein-1 (MCP-1) in mammal is reduced It is flat；And/or (ii) improves leptin resistance, the sensitiveness in vivo to Leptin is improved, wherein, the heavy wall mushroom probiotics is selected from The following group：Association fecal bacteria (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister invisus) or It is combined.
5. purposes as claimed in claim 4, it is characterised in that the composition or preparation are also independently or additionally used to select From one or more purposes of the following group：
(iii) body weight increase of mammal is suppressed；
(iv) the body fat ratio (the ratio between fat weight/body weight) of mammal is reduced；
(v) blood lipid level of mammal is reduced；
(vi) level of the HDL (HDLC) in mammal is improved；
(vii) low-density lipoprotein (LDLC) level in reduction mammal.
6. a kind of be used for the composition for the treatment of and/or pre- preventing obesity, the composition includes：(i) firmicutes of safe and effective amount Class probiotics；On (ii) food or pharmaceutically acceptable carrier；Wherein, the heavy wall mushroom probiotics is selected from the group：Companion Raw fecal bacteria (Coprococcus eutactus), muddiness wear Alister bacterium (Dialister invisus) or its combination.
7. composition as claimed in claim 6, it is characterised in that the composition contains 1 × 10-1 × 1020Cfu/mL or Cfu/g heavy wall mushroom probiotics, preferably 1 × 104-1×1015Cfu/mL or cfu/g heavy wall mushroom probiotics, by described The cumulative volume or gross weight meter of composition.
8. composition as claimed in claim 6, it is characterised in that in described composition, containing 0.0001-99wt%, compared with Heavy wall mushroom probiotics described in good ground 0.1-90wt%, with the gross weight meter of the composition.
9. composition as claimed in claim 6, it is characterised in that described composition also contains other probiotics and/or benefit Raw member.
10. the preparation method of composition described in a kind of claim 6, including step：
Mixed by (i) heavy wall mushroom probiotics, with acceptable carrier or pharmaceutically acceptable carrier on (ii) food, so that Form the composition described in claim 6.
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