Probiotic composition for alleviating sarcopenia obesity and application thereof

Abstract

The present invention relates to the field of biotechnology and discloses a probiotic composition for alleviating sarcopenic obesity and its use. The composition comprises at least two of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis; wherein the Lactobacillus plantarum has a deposit number of CGMCC No. 25839; the Lactobacillus rhamnosus has a deposit number of CGMCC No. 26968; and the Bifidobacterium animalis subsp. lactis has a deposit number of CGMCC No. 27998. The composition of the present invention can increase muscle mass, improve muscle tissue morphology, increase adenosine triphosphate, and inhibit abnormal weight gain, thereby effectively preventing and treating sarcopenic obesity.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a probiotic composition for alleviating sarcopenia obesity and an application thereof. Background Art

[0002] Sarcopenic obesity (SO) is a chronic disease characterized by the coexistence of obesity and sarcopenia (hereafter referred to as sarcopenia). Sarcopenia is a disorder of the skeletal muscle system characterized by decreased muscle mass, strength, and function. The pathogenesis of sarcopenic obesity is complex, including aging, an inappropriate lifestyle (sedentary lifestyle, poor diet, and lack of exercise), inflammation, and comorbidities of acute and chronic diseases. This dual pressure of muscle loss and fat accumulation can lead to complications such as frailty, disability, falls, fractures, metabolic diseases, and cancer, resulting in an increasing mortality rate. With the further aging of the population and the increasing number of obese people, the incidence of sarcopenic obesity continues to rise, making it a major global public health issue.

[0003] Currently, treatments for sarcopenic obesity primarily include dietary intervention, exercise therapy, and medication. Dietary intervention requires patients to strictly control their diet and caloric intake, making long-term adherence difficult. Exercise therapy requires a certain level of physical condition and exercise capacity, making it difficult for some elderly or frail patients to implement. Existing medications also have significant side effects and uncertain efficacy.

[0004] Therefore, developing a safe, effective and easy-to-use product to alleviate sarcopenia obesity is of great practical significance. Summary of the Invention

[0005] The present invention aims to, at least to some extent, address at least one of the technical problems existing in the prior art. The present invention proposes a composition that can effectively alleviate sarcopenic obesity. The composition comprises at least two of three probiotics: Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis. After in-depth research, the inventors discovered that the composition of the present invention has multiple benefits in improving sarcopenic obesity. It can increase muscle mass, improve muscle tissue morphology, and thus enhance muscle functionality. Simultaneously, the composition can effectively increase adenosine triphosphate (ATP) levels, providing more energy and thus improving exercise capacity. Furthermore, it can effectively inhibit abnormal weight gain, thereby achieving comprehensive improvements in sarcopenic obesity.

[0006] In a first aspect of the present invention, the present invention proposes a composition. According to an embodiment of the present invention, the composition includes: at least two of plant lactobacillus, rhamnosus lactobacillus and animal bifidobacterium lactis subspecies; wherein the preservation unit of the plant lactobacillus is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is September 28, 2022, and the preservation number is CGMCC No.25839; the preservation unit of the rhamnosus lactobacillus is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is March 31, 2023, and the preservation number is CGMCC No.26968; the preservation unit of the animal bifidobacterium lactis subspecies is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is July 24, 2023, and the preservation number is CGMCC No.27998.

[0007] According to an embodiment of the present invention, the composition may further include at least one of the following additional technical features:

[0008] According to an embodiment of the present invention, the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis are independently live bacteria, inactivated bacteria or a combination thereof.

[0009] According to an embodiment of the present invention, the total effective viable count of the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis is not less than 1×10 6 CFU / g.

[0010] According to an embodiment of the present invention, the ratio of the viable cell counts of the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis is (1-5):(1-5):(1-5).

[0011] In a second aspect of the present invention, the present invention provides a product. According to an embodiment of the present invention, the product comprises the composition described in the first aspect.

[0012] According to an embodiment of the present invention, the product may further include at least one of the following additional technical features:

[0013] According to an embodiment of the present invention, the product comprises a medicine.

[0014] According to an embodiment of the present invention, the drug further includes a pharmaceutically acceptable carrier.

[0015] In the third aspect of the present invention, the present invention proposes the use of the composition described in the first aspect or the product described in the second aspect in the preparation of a medicament for preventing and treating sarcopenia obesity.

[0016] In the third aspect of the present invention, the present invention provides an application of the composition of the first aspect or the product of the second aspect, wherein the application is selected from at least one of the following:

[0017] Application in the preparation of preparations for increasing muscle mass;

[0018] Application in the preparation of preparations for improving athletic performance;

[0019] Application in the preparation of preparations for increasing ATP content;

[0020] Application in the preparation of preparations for improving muscle tissue morphology;

[0021] Application in the preparation of preparations for reducing body weight.

[0022] After extensive experiments, the inventors discovered that the combined use of three probiotics—Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis—can effectively alleviate sarcopenic obesity. Therefore, a composition or product containing these three probiotics can increase muscle mass, improve muscle tissue morphology, and increase adenosine triphosphate (ATP), while effectively inhibiting abnormal weight gain, thereby effectively preventing and treating sarcopenic obesity.

[0023] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0025] Figure 1 This is a graph showing the experimental results of the probiotic powder according to Example 1 of the present invention for improving sarcopenia (total exercise distance), compared with the model control group. * P <0.05,** P <0.01,*** P <0.001;

[0026] Figure 2 This is a graph showing the experimental results of the probiotic powder in Example 2 of the present invention on improving sarcopenia (ATP content). Compared with the model control group, * P <0.05,** P <0.01,*** P <0.001;

[0027] Figure 3 This is a typical image of zebrafish muscle tissue pathological section after sample treatment according to Example 3 of the present invention;

[0028] Figure 4This is a graph showing the effect of the composite probiotics on weight gain in mice according to Example 4 of the present invention. Compared with the model control group, ** P <0.01. DETAILED DESCRIPTION

[0029] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0030] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing embodiments and examples and are not intended to limit the present invention.

[0031] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0032] In the present invention, "plurality", "multiple", "multiple times", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0033] As used herein, "combination thereof", "any combination thereof", "any combination thereof" and the like include all suitable combinations of any two or more of the listed items.

[0034] In this document, the terms “contain”, “include” or “include” are open expressions, that is, they include the contents specified in the present invention but do not exclude other contents.

[0035] In the present invention, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​within the numerical interval is considered continuous and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval refers only to integers within the numerical interval, including the two endpoint integers of the numerical range, and each integer between the two endpoints, is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all subranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows for broadly including quantitative intervals such as percentage intervals, ratio intervals, and ratio intervals.

[0036] In the present invention, when referring to the unit of a data range, if the unit is only after the right endpoint, it means that the units of the left endpoint and the right endpoint are the same. For example, 3-5h means that the units of the left endpoint "3" and the right endpoint "5" are both h (hours).

[0037] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0038] Loss of skeletal muscle mass and function is common in obese individuals due to dyslipidemia, acute and chronic comorbidities, and aging-related metabolic changes. Excessive obesity accompanied by reduced muscle mass or function is termed sarcopenic obesity (SO). This condition portends adverse clinical outcomes, necessitating the development of effective strategies for the prevention and treatment of SO.

[0039] Probiotics, a class of active microorganisms that benefit the host, have garnered widespread attention in recent years for improving human metabolic health. Studies have found that an imbalance in the gut microbiome is closely associated with the development and progression of sarcopenic obesity, suggesting that regulating the gut microbiome may offer a new approach to alleviating sarcopenic obesity. However, a highly effective probiotic combination specifically targeting sarcopenic obesity has yet to be developed.

[0040] In light of this, the inventors conducted research on various probiotics and discovered that the combined use of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis exhibits multiple benefits in improving sarcopenic obesity. These benefits include increasing muscle mass, improving muscle morphology, and thereby enhancing muscle functionality. Furthermore, they effectively increase adenosine triphosphate (ATP) levels, providing more energy and improving athletic performance. Furthermore, they effectively inhibit abnormal weight gain, achieving comprehensive improvements in sarcopenic obesity. The following details the composition, product, and applications containing these three bacteria.

[0041] Composition

[0042] In a first aspect of the present invention, the present invention proposes a composition. According to an embodiment of the present invention, the composition includes at least two of plant lactobacillus, rhamnosus lactobacillus and animal bifidobacterium lactis subspecies; wherein the preservation unit of the plant lactobacillus is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is September 28, 2022, and the preservation number is CGMCC No.25839; the preservation unit of the rhamnosus lactobacillus is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is March 31, 2023, and the preservation number is CGMCC No.26968; the preservation unit of the animal bifidobacterium lactis subspecies is the General Microbiology Center of the China Culture Collection of Microorganisms, and the preservation time is July 24, 2023, and the preservation number is CGMCC No.27998.

[0043] After extensive experiments, the inventors discovered that the combined use of three probiotics—Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis—can effectively alleviate sarcopenic obesity. Therefore, a composition containing these three probiotics can have multiple benefits in improving sarcopenic obesity. It can increase muscle mass, improve muscle morphology, and thus enhance muscle functionality. Simultaneously, the composition can effectively increase adenosine triphosphate (ATP) levels, providing more energy and improving athletic ability. Furthermore, it can effectively inhibit abnormal weight gain, thereby achieving comprehensive improvements in sarcopenic obesity.

[0044] It should be noted that the "Lactobacillus plantarum," "Lactobacillus rhamnosus," and "Bifidobacterium animalis subsp. lactis" described in the present invention are all known and disclosed strains. Specifically, relevant information about the "Lactobacillus plantarum" described in the present invention has been clearly disclosed in patent CN119372076A, relevant information about the "Lactobacillus rhamnosus" described in the present invention has been clearly disclosed in patent CN117126788A, and relevant information about the "Bifidobacterium animalis subsp. lactis" described in the present invention has been clearly disclosed in patent CN117448198A.

[0045] In some embodiments of the present invention, the Lactobacillus plantarum may be a living bacterium, or a Lactobacillus plantarum that has been inactivated, genetically recombined, transformed or modified, attenuated, chemically treated, or physically treated and retains all or part of its original biological activity, or a lysate, culture (e.g., supernatant) of the bacterium, or a component extracted from the culture.

[0046] In some embodiments of the present invention, the Lactobacillus rhamnosus may be a living bacterium, or a Lactobacillus rhamnosus that has been inactivated, genetically recombined, transformed or modified, attenuated, chemically treated, or physically treated and retains all or part of its original biological activity, or a lysate, culture (e.g., supernatant) of the bacterium, or a component extracted from the culture.

[0047] In some embodiments of the present invention, the animal Bifidobacterium lactis subspecies can be a living bacterium, or it can be an animal Bifidobacterium lactis subspecies that has been inactivated, genetically recombined, transformed or modified, attenuated, chemically treated, or physically treated to fully or partially retain its original biological activity, or it can be a bacterial lysate, culture (such as supernatant) or a component extracted from the culture.

[0048] In some embodiments of the present invention, the total effective viable count of the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis is not less than 1×10 6 CFU / g. For example, it can be 1×10 6 CFU / g, 1×10 7 CFU / g, 1×10 8 CFU / g, 1×10 9 CFU / g, 1×10 10 CFU / g, 1×10 11 CFU / g, 1×10 12 CFU / g, etc., or can be within a range consisting of any of the above values. Thus, the composition can effectively increase the body's muscle content, improve muscle tissue morphology, increase adenosine triphosphate (ATP) content, inhibit abnormal weight gain, and thus effectively prevent and treat sarcopenia and obesity.

[0049] In some embodiments of the present invention, the ratio of the viable counts of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis is (1-5):(1-5):(1-5). For example, the ratio may be 1:1:1, 1:1:2, 1:3:1, 4:1:2, 5:1:5, etc., or may be within any range of the aforementioned values. Thus, by ensuring that the ratio of the viable counts of the three probiotics is within the aforementioned range, the composition can effectively increase muscle mass, improve muscle tissue morphology and adenosine triphosphate (ATP) content, and inhibit abnormal weight gain, thereby effectively preventing and treating sarcopenia and obesity.

[0050] product

[0051] In its second aspect, the present invention provides a product. According to an embodiment of the present invention, the product comprises the composition described in the first aspect. As previously described, the composition of the present invention can effectively increase muscle mass, improve muscle tissue morphology, increase adenosine triphosphate (ATP) levels, and inhibit abnormal weight gain, thereby effectively preventing and treating sarcopenia-related obesity. Therefore, the product of the present invention can effectively prevent and treat sarcopenia-related obesity.

[0052] In some embodiments of the invention, the product comprises a pharmaceutical.

[0053] In some embodiments of the present invention, the drug is a pharmaceutical.

[0054] For purposes of this invention, "drug" includes any agent, compound, composition, or mixture that provides a pharmacological effect in vivo or in vitro, often with a beneficial effect. The scope of the pharmacological effect of a "drug" in vivo is not particularly limited and may be systemic or localized. The activity of the "drug" is not particularly limited and may be an active substance that interacts with other substances or an inert substance that does not interact.

[0055] In the present invention, a "drug" is a directly administrable pharmaceutical preparation, which usually has a prescribed usage and dosage.

[0056] In some embodiments of the present invention, the drug further comprises a pharmaceutically acceptable carrier.

[0057] In the present invention, "pharmaceutically acceptable" refers to those ligands, materials, compositions and / or dosage forms that are suitable for administration to a patient within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0058] In the present invention, "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. As used herein, the phrase "pharmaceutically acceptable carrier" includes buffers, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with drug administration. Each carrier must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients in the formulation and not harmful to the patient. Suitable examples include, but are not limited to: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch, potato starch, and substituted or unsubstituted β-cyclodextrins; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil. oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer; and (21) other nontoxic compatible substances used in pharmaceutical formulations.

[0059] In some embodiments of the present invention, the dosage form of the drug is a tablet, capsule, granule, pill, ointment, solution, suspension, emulsion, cream, spray, drop, patch or tube feeding preparation.

[0060] In some embodiments of the present invention, the drug may further include other active pharmaceutical ingredients. The other active pharmaceutical ingredients may be pharmaceutical ingredients with sarcopenia prevention and treatment effects, or pharmaceutical ingredients with obesity prevention and treatment effects, or pharmaceutical ingredients with other disease prevention and treatment effects.

[0061] Applications and Methods

[0062] In the third aspect of the present invention, the present invention proposes the use of the composition described in the first aspect or the product described in the second aspect in the preparation of a medicament for preventing and treating sarcopenia obesity.

[0063] In the present invention, "preventing and / or treating" and "preventing and / or treating" have the same meaning and can be used interchangeably. In the present invention, "preventing and treating" includes aspects such as prevention, treatment, and adjuvant therapy. As used herein, "preventing and treating" means alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition. "Preventing and treating" also includes curing, preventing the development of, or alleviating to a certain extent, one or more symptoms of a disease or condition.

[0064] In the fourth aspect of the present invention, the present invention proposes the use of the composition described in the first aspect or the product described in the second aspect, and the use is selected from at least one of the following: use in preparing a preparation for increasing muscle content; use in preparing a preparation for improving athletic ability; use in preparing a preparation for increasing ATP content; use in preparing a preparation for improving muscle tissue morphology; use in preparing a preparation for reducing body weight.

[0065] In some embodiments of the present invention, in the above applications, the preparation includes a pharmaceutical preparation or a food preparation. It should be noted that the composition or product can be prepared into food and medicine.

[0066] In some embodiments, the pharmaceutical preparation can be a liquid preparation or a solid preparation. Liquid preparations refer to preparations containing a liquid phase, and non-limiting examples include solutions, suspensions, emulsions, etc. Non-limiting examples of solid preparations include tablets, capsules, granules, pills, etc.

[0067] In some embodiments, depending on the mode of administration, the pharmaceutical preparation can be an oral preparation, an injection, a drop, a patch, a tube feeding preparation, etc.

[0068] In the fifth aspect of the present invention, the present invention proposes a method for preventing and treating sarcopenic obesity, characterized in that it comprises: administering a therapeutically effective amount of the composition described in the first aspect or the product described in the second aspect to a subject.

[0069] In the present invention, a "subject" is an animal, preferably a mammal, more preferably a human. Subjects include, but are not limited to, consumers of health products and patients with diseases, conditions, and / or symptoms. The subject in the present invention is preferably a mammal. The term "mammal" primarily refers to warm-blooded vertebrate mammals, including, but not limited to, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice (e.g., rats, mice), pigs, cattle, sheep, horses, and humans, preferably primates, and more preferably humans.

[0070] In the present invention, "therapeutically effective amount" refers to the amount of the active ingredient of the drug that will cause a biological or medical response in an individual to a disease, disorder and / or symptom, for example, the amount of the composition or product of the present invention that brings a pharmacologically positive effect to an individual, and the pharmacologically positive effect includes but is not limited to reducing or inhibiting the activity of an enzyme or protein or improving symptoms, alleviating symptoms, slowing or delaying the progression of a disease or preventing a disease, etc.

[0071] The following embodiments of the present invention are described in detail. The embodiments described below are exemplary and are only used to explain the present invention, and are not to be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments, the techniques or conditions described in the literature in this field or the product specifications are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be obtained commercially.

[0072] The VB165 strain involved in the embodiment of the present invention is classified and named as Lactobacillus plantarum ( Lactobacillus plantarum ), the depository is the General Microbiology Center of China Culture Collection Administration, the deposit date is September 28, 2022, the deposit number is CGMCC No.25839, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0073] The VB255 strain involved in the embodiment of the present invention is classified and named as Lactobacillus rhamnosus ( Lactobacillus rhamnosus ), the depository is the General Microbiology Center of China Culture Collection Administration, the deposit date is March 31, 2023, the deposit number is CGMCC No.26968, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0074] The VB301 strain involved in the embodiment of the present invention is classified as Bifidobacterium animalis subsp. lactis ( Bifidobacterum animalis subsp. lactis ), the depository is the General Microbiology Center of China Culture Collection Administration, the deposit date is July 24, 2023, the deposit number is CGMCC No.27998, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0075] The BMZ140526 strain (Bifidobacterium animalis) involved in the embodiments of the present invention was purchased from Ningbo Mingzhou Biotechnology Co., Ltd.

[0076] The zebrafish involved in the embodiments of the present invention were all raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450~550 μS / cm; the pH was 6.5~8.5; the hardness was 50~100 mg / LCaCO3), and were bred and provided by the Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004, and the breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2025-11001-01.

[0077] Example 1: Evaluation of the efficacy of improving sarcopenia (behavioral)

[0078] Wild-type AB zebrafish (3 days post fertilization, 3 DPF) were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well. The zebrafish in different wells were randomly divided into a normal control group, a positive control group, a model control group, and five experimental groups: the probiotic VB165+VB255 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB255+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB255+VB301 group (a 1:1 ratio of viable bacteria between the three bacteria), and the probiotic VB165+VB301+BMZ140526 group (a 1:1:1 ratio of viable bacteria between the three bacteria).

[0079] The normal control group was given 3 mL of fish culture water. Each experimental group was given 1×10 probiotic combination powder dissolved in fish culture water. 7 CFU / mL, the positive control group was given 62.5 μg / mL of whey protein powder dissolved in fish water, and the capacity of each well was 3mL. In addition, except for the normal control group, 2μmol / L dexamethasone was added to the fish water in the other groups to establish a zebrafish sarcopenia model, that is, the fish water in the experimental group simultaneously dissolved probiotic combination powder and dexamethasone, the fish water in the positive control group simultaneously dissolved whey protein powder and dexamethasone, and the fish water in the model control group only dissolved dexamethasone. The medium was changed every day. After 4 days of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and transferred to a 96-well plate, 1 tail / well, and 200 μL / well of fish water were given. The total movement distance of the zebrafish within 1 hour was measured using the zebrafish behavior analysis system, and the statistical analysis results of this indicator were used to evaluate the efficacy of the sample in improving sarcopenia. The statistical processing results are expressed as mean ± SE. Statistical analysis was performed using SPSS26.0 software, and p<0.05 indicated that the difference was statistically significant. The results are as follows Figure 1As shown in the figure, compared with the model control group, the probiotic composition has the effect of improving sarcopenia, which is specifically manifested in increasing the total exercise distance, among which the VB165+VB255+VB301 group has a better effect.

[0080] Example 2: Evaluation of Sarcopenia Improvement Efficacy (ATP Content)

[0081] Wild-type AB zebrafish (3 days post fertilization, 3 DPF) were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well. The zebrafish in different wells were randomly divided into a normal control group, a positive control group, a model control group, and five experimental groups: the probiotic VB165+VB255 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB255+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB255+VB301 group (a 1:1 ratio of viable bacteria between the three bacteria), and the probiotic VB165+VB301+BMZ140526 group (a 1:1:1 ratio of viable bacteria between the three bacteria).

[0082] The normal control group was given 3 mL of fish culture water. Each experimental group was given 1×10 probiotic combination powder dissolved in fish culture water. 7 CFU / mL, the positive control group was given 62.5 μg / mL of whey protein powder dissolved in fish water, and the capacity of each well was 3mL. In addition, except for the normal control group, 2μmol / L dexamethasone was added to the fish water in the other groups to establish a zebrafish sarcopenia model, that is, the fish water in the experimental group simultaneously dissolved probiotic combination powder and dexamethasone, the fish water in the positive control group simultaneously dissolved whey protein powder and dexamethasone, and the fish water in the model control group only dissolved dexamethasone. The medium was changed every day, and after 4 days of treatment at 28°C, data was collected using a multifunctional microplate reader according to the operating method of the ATP (adenosine triphosphate) content determination kit, and the ATP content in the zebrafish was analyzed. The statistical analysis results of this indicator were used to evaluate the efficacy of the sample in improving sarcopenia. The statistical processing results are expressed as mean ± SE. Statistical analysis was performed using SPSS26.0 software, and p<0.05 indicated that the difference was statistically significant. The results are as follows Figure 2 As shown in the figure, compared with the model control group, the probiotic composition can improve sarcopenia, specifically by increasing the ATP content, among which the VB165+VB255+VB301 group has a better effect.

[0083] Example 3: Evaluation of efficacy in improving sarcopenia (muscle histopathology)

[0084] Wild-type AB zebrafish (3 days post fertilization, 3 DPF) were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well. The zebrafish in different wells were randomly divided into a normal control group, a positive control group, a model control group, and five experimental groups: the probiotic VB165+VB255 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB255+VB301 group (a 1:1 ratio of viable bacteria between the two bacteria), the probiotic VB165+VB255+VB301 group (a 1:1 ratio of viable bacteria between the three bacteria), and the probiotic VB165+VB301+BMZ140526 group (a 1:1:1 ratio of viable bacteria between the three bacteria).

[0085] The normal control group was given 3 mL of fish culture water. Each experimental group was given 1×10 probiotic combination powder dissolved in fish culture water. 7 CFU / mL, the positive control group was given 62.5 μg / mL of whey protein powder dissolved in fish water, and the capacity of each well was 3mL. In addition, except for the normal control group, 2μmol / L dexamethasone was added to the fish water in the other groups to establish a zebrafish sarcopenia model, that is, the fish water in the experimental group simultaneously dissolved probiotic combination powder and dexamethasone, the fish water in the positive control group simultaneously dissolved whey protein powder and dexamethasone, and the fish water in the model control group only dissolved dexamethasone. The medium was changed every day, and after 4 days of treatment at 28°C, the zebrafish in each group underwent fixation, dehydration, embedding, sectioning, H&E staining and other steps, and muscle tissue pathology examination was performed to evaluate the efficacy of the samples in improving sarcopenia. The results are as follows Figure 3 As shown in the figure, the zebrafish muscle tissue in the normal control group showed normal morphology, with thin, long muscle cells arranged neatly and tightly. The muscle tissue in the model control group was loose and irregularly arranged, indicating that the model was successfully established. The zebrafish muscle tissue in the positive control whey protein powder concentration group of 62.5 μg / mL was similar to that of the normal control group and had a regular arrangement, indicating that whey protein powder has the effect of improving sarcopenia. The zebrafish muscle tissue morphology in each probiotic combination group was significantly improved compared to the model control group, with a regular arrangement, indicating that the probiotic combination has the effect of improving sarcopenia, among which the VB165+VB255+VB301 group had the best effect.

[0086] Example 4: Effect of compound probiotics on weight gain in older mice

[0087] Healthy male C57BL / 6J mice aged 6 months were housed in a temperature of 20-26°C, humidity of 40%-70%, with light and dark alternation every 12 h, and were given free access to food. They were adaptively housed for 7 days.

[0088] Mice with similar body weight were randomly divided into 7 groups, with 10 mice in each group, namely normal control group, model group, probiotic VB165+VB255 group (the ratio of the number of live bacteria of the two bacteria was 1:1), probiotic VB165+VB301 group (the ratio of the number of live bacteria of the two bacteria was 1:1), probiotic VB255+VB301 group (the ratio of the number of live bacteria of the two bacteria was 1:1), probiotic VB165+VB255+VB301 group (the ratio of the number of live bacteria of the three bacteria was 1:1:1), and probiotic VB165+VB301+BMZ140526 group (the ratio of the number of live bacteria of the three bacteria was 1:1:1).

[0089] The normal control group was given a normal diet and free access to water. The model group was given a high-fat diet and free access to water to induce obesity. The probiotic combination group was given a high-fat diet and free access to water. The control and model groups were gavaged daily with 400 μL of deionized water; the probiotic combination group was gavaged daily with 400 μL of a probiotic combination powder solution suspended in deionized water, with a total viable count of 1 × 10 probiotics. 9 CFU. The intervention lasted for 18 weeks. Every three weeks, mice were weighed using an electronic balance. Weight changes were recorded and plotted to observe weight gain and loss trends in different groups.

[0090] The results of mouse weight changes are as follows Figure 4 As shown in the figure, during the experiment, the weight of the mice in the normal control group remained basically unchanged, maintained at about 27g, and their physical signs were good. The mice in the model group and the probiotic experimental group continued to grow normally, became obese, and their weight was significantly higher than that of the normal control group. Compared with the model group, the weight gain of the mice in the probiotic experimental group was less than that of the model group, but this difference was not significant. Among them, the weight gain of the mice in the VB165+VB255+VB301 group was significantly lower than that of the model group at the 15th and 18th weeks after the probiotic solution was given for 12 weeks. The results show that feeding mice with the VB165+VB255+VB301 compound probiotic powder for more than 12 weeks will have a significant inhibitory effect on the weight gain of mice fed a high-fat diet, showing good weight loss potential.

[0091] In the description of this specification, the reference terms "one embodiment," "some embodiments," "example," "specific example," or "some examples" mean that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0092] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A composition comprising probiotics, characterized in that By Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) and Bifidobacterium animalis subsp. lactis ( Bifidobacterium animalis subsp. lactis )composition; The Lactobacillus plantarum strain was deposited at the General Microbiology Center of the China Culture Collection Administration Committee on Microorganisms on September 28, 2022, with a deposit number of CGMCC No. 25839. The Lactobacillus rhamnosus strain is deposited at the General Microbiology Center of the China Culture Collection Administration, with a deposit date of March 31, 2023, and a deposit number of CGMCC No. 26968. The depository of the animal Bifidobacterium lactis subsp. lactis is the General Microbiology Center of the China Culture Collection Administration of Microorganisms, with a deposit date of July 24, 2023, and a deposit number of CGMCC No. 27998. The ratio of the viable bacterial counts of the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis is (1-5):(1-5):(1-5).

2. The composition according to claim 1, characterized in that The Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis are each independently a live bacterium.

3. The composition according to claim 2, characterized in that The total effective viable count of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subspecies lactis is not less than 1×10 6 CFU / g.

4. A product comprising a probiotic composition, characterized in that The product comprises the composition according to any one of claims 1 to 3.

5. The product according to claim 4, characterized in that The products include pharmaceuticals.

6. The product according to claim 5, characterized in that The drug further includes a pharmaceutically acceptable carrier.

7. Use of the composition according to any one of claims 1 to 3 or the product according to any one of claims 4 to 6 in the preparation of a medicament for preventing and treating sarcopenia obesity.

8. Use of the composition according to any one of claims 1 to 3 or the product according to any one of claims 4 to 6, wherein the use is selected from at least one of the following: Application in the preparation of preparations for increasing muscle mass; Application in the preparation of preparations for improving athletic performance; Application in the preparation of preparations for increasing ATP content; Application in the preparation of preparations for improving muscle tissue morphology; Application in the preparation of preparations for reducing body weight.

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