Probiotic composition for regulating blood sugar metabolism and application thereof

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to a probiotic composition for regulating blood sugar metabolism and its application. The probiotic composition comprises the following components: two or three of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis lactis subspecies. The present invention provides a probiotic composition, which comprises two or three of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis lactis subspecies, and further optimizes the synergistic effect of the strains by adjusting the type collocation and the ratio of viable bacteria count of the strains, so that the probiotic composition has better effects in preventing and treating diabetes, controlling blood sugar levels, regulating intestinal flora, and anti-inflammatory. In addition, the probiotic composition can also be used to prevent, alleviate, assist in the treatment or treatment of diabetes, providing a more effective and safer solution for the management of diabetes.

Description

Technical Field

[0001] The present invention belongs to the field of probiotics, and in particular relates to a probiotic composition for regulating blood sugar metabolism and an application thereof. Background Art

[0002] Diabetes is a metabolic disease characterized by chronically elevated blood sugar levels. Long-term high blood sugar levels lead to chronic damage or dysfunction in various tissues of the body, particularly the eyes, kidneys, heart, blood vessels, and nerves. As a global public health issue, its incidence has shown a significant upward trend in recent years, placing a heavy economic burden on patients and their families. Currently, the prevention and treatment of diabetes primarily rely on medication, diet, and exercise. However, these methods still have numerous shortcomings. Medication can cause side effects such as hypoglycemia and gastrointestinal reactions; while the effectiveness of diet and exercise varies from person to person and can be difficult to maintain long-term.

[0003] Relevant studies have found that intestinal flora is closely related to diabetes, and regulating intestinal flora has become a new direction in diabetes treatment research. Some probiotic strains have been found to have positive effects on the prevention and treatment of diabetes, but existing probiotic products have problems such as single formula and limited effect.

[0004] Therefore, developing a probiotic composition that can effectively prevent or treat diabetes is of great significance for improving the prognosis of diabetic patients and reducing the medical costs of diabetic patients. Summary of the Invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention provides a probiotic composition for regulating blood sugar metabolism and its application. The present invention provides a probiotic composition comprising two or more of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis lactis subsp. lactis; wherein the Lactobacillus plantarum is Lactobacillus plantarum VB165, whose preservation number is CGMCC No. 25839; the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, whose preservation number is CGMCC No. 26968; and the Bifidobacterium animalis lactis subsp. lactis VB301, whose preservation number is CGMCC No. 27998. By adjusting the type combination and the ratio of viable bacteria of the strains, the synergistic effect of the strains is further optimized, so that the probiotic composition has better effects in preventing and treating diabetes, controlling blood sugar levels, regulating intestinal flora and anti-inflammatory. In addition, the probiotic composition can also be used to prevent, alleviate, assist in the treatment or treatment of diabetes, providing a more effective and safer solution for diabetes management.

[0006] In a first aspect of the present invention, a probiotic composition is provided. According to an embodiment of the present invention, the probiotic composition comprises the following components: two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis; wherein the Lactobacillus plantarum is Lactobacillus plantarum ( Lactobacillus plantarum ) VB165, its deposit number is CGMCC No.25839, deposit date: September 28, 2022, deposit unit: General Microbiology Center of China Culture Collection Administration; deposit unit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the rhamnosus Lactobacillus is rhamnosus Lactobacillus ( Lactobacillus rhamnosus )VB255, its deposit number is CGMCC No.26968; deposit date: March 31, 2023, deposit unit: General Microbiology Center of China Culture Collection Administration; deposit unit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the animal Bifidobacterium lactis subsp. animalis subsp. lactis ( Bifidobacterium animalis subsp. lactis ) VB301, deposited with CGMCC No. 27998; date of deposit: July 24, 2023; deposited at: General Microbiology Center, China Culture Collection Administration; address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The probiotic composition according to an embodiment of the present invention comprises two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis. The synergistic action of these strains effectively regulates intestinal flora, enhances intestinal barrier function, and reduces inflammatory responses, thereby demonstrating significant effectiveness in preventing and treating diabetes, controlling blood sugar levels, and combating inflammation, and has broad application prospects.

[0007] According to an embodiment of the present invention, the above-mentioned probiotic composition may also have the following additional technical features:

[0008] According to an embodiment of the present invention, the Lactobacillus plantarum and the Lactobacillus rhamnosus are contained; the ratio of the number of viable cells of the Lactobacillus plantarum to the number of viable cells of the Lactobacillus rhamnosus is (1-5):(1-5).

[0009] According to an embodiment of the present invention, the Lactobacillus plantarum and the Bifidobacterium animalis subspecies lactis are contained; the ratio of the number of viable bacteria of the Lactobacillus plantarum and the Bifidobacterium animalis subspecies lactis is (1-5):(1-5).

[0010] According to an embodiment of the present invention, the composition comprises the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis; the ratio of the number of viable bacteria of the Lactobacillus rhamnosus to the number of viable bacteria of the Bifidobacterium animalis subspecies lactis is (1-5):(1-5).

[0011] According to an embodiment of the present invention, the composition comprises the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis; the ratio of the number of viable cells of the Lactobacillus plantarum, the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis is (1-5):(1-5):(1-5).

[0012] In a second aspect, the present invention provides a composite bacterial agent. According to an embodiment of the present invention, the active ingredient of the composite bacterial agent is the probiotic composition described in the first aspect. The composite bacterial agent according to an embodiment of the present invention, by incorporating the aforementioned probiotic composition into the composite bacterial agent, makes the probiotic composition easier to store, transport, and use, further improving its convenience and practicality, further expanding its scope of application, and facilitating use in different scenarios.

[0013] In a third aspect, the present invention provides for the use of the probiotic composition described in the first aspect or the composite bacterial agent described in the second aspect in the preparation of food, health products, or medicines. According to embodiments of the present invention, the medicine has at least one of the following uses: preventing, alleviating, assisting in the treatment, and / or treating diabetes; controlling blood sugar levels; or providing anti-inflammatory benefits. According to embodiments of the present invention, by applying the aforementioned probiotic composition to food, health products, or medicines, particularly medicines, a safer and more effective treatment option is provided for diabetic patients.

[0014] In the fourth aspect of the present invention, the present invention proposes the use of the probiotic composition described in the first aspect or the composite bacterial agent described in the second aspect in the preparation of a product for preventing, alleviating, assisting in the treatment or curing diabetes.

[0015] It will be understood by those skilled in the art that the features and advantages described above for the probiotic composition or compound bacterial agent are also applicable to this application and will not be repeated here.

[0016] 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

[0017] 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:

[0018] Figure 1 The fasting blood glucose measurement results of each experimental mouse before and after the intervention of probiotic compositions I to V in Example 1 of the present invention are shown in FIG, wherein NS indicates that the difference in the results of this part of the data is not significant, and * indicates P <0.05, ** P <0.01, *** P <0.001;

[0019] Figure 2 This is a graph showing the results of glucose tolerance tests on experimental mice after intervention with probiotic compositions I to V in Example 1 of the present invention;

[0020] Figure 3 The inflammatory factor detection results of each experimental mouse after intervention with probiotic compositions I to V in Example 1 of the present invention are shown in Figure A, wherein A is the inflammatory factor IL-6 detection result of each experimental mouse after intervention with probiotic compositions I to V, and B is the inflammatory factor IL-1β detection result of each experimental mouse after intervention with probiotic compositions I to V, wherein * is P <0.05, ** P <0.01, *** P <0.001. DETAILED DESCRIPTION

[0021] 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.

[0022] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0023] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0024] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention, but not excluding other contents.

[0025] As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0026] Probiotic composition

[0027] The present invention proposes a probiotic composition. According to an embodiment of the present invention, the probiotic composition comprises the following components: two or more of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis lactis subspecies; wherein the Lactobacillus plantarum is Lactobacillus plantarum VB165, and its preservation number is CGMCC No.25839; the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, and its preservation number is CGMCC No.26968; the Bifidobacterium animalis lactis subspecies VB301, and its preservation number is CGMCC No.27998. The probiotic composition according to an embodiment of the present invention comprises two or more of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis lactis subspecies. Under the synergistic action of these strains, it can better and more effectively regulate the intestinal flora, enhance the intestinal barrier function, and reduce the inflammatory response, thereby showing significant effects in preventing and treating diabetes, controlling blood sugar levels and anti-inflammation, and has broad application prospects.

[0028] According to an embodiment of the present invention, the Lactobacillus plantarum is Lactobacillus plantarum VB165, which has a deposit number of CGMCC No. 25839. Thus, by optimizing the strain source of Lactobacillus plantarum, an important basis is provided for the subsequent standardized production and quality control of the probiotic composition. In addition, the strain itself can effectively inhibit the activity of α-glucosidase, while also having the activity of degrading bile salts and inhibiting major intestinal pathogens, and has good tolerance to gastric acid and choline.

[0029] According to an embodiment of the present invention, the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, which has a deposit number of CGMCC No. 26968. Thus, by optimizing the strain source of Lactobacillus rhamnosus, an important basis is provided for the subsequent standardized production and quality control of the probiotic composition. In addition, the strain itself has good stability and can be stably present in pancreatic juice without affecting the intestinal microecological environment. It also has a good inhibitory effect on Candida albicans and / or Shigella.

[0030] According to an embodiment of the present invention, the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis VB301, which has a deposit number of CGMCC No. 27998. Thus, by optimizing the strain source of Bifidobacterium animalis subsp. lactis, an important basis is provided for the subsequent standardized production and quality control of the probiotic composition. In addition, the strain itself has a significant bile salt-lowering ability and has strong intestinal tolerance and antibacterial ability.

[0031] It should be noted that the plantarum lactobacillus, rhamnosus lactobacillus and animal bifidobacterium lactis subspecies of the probiotic composition of the present invention do not only include plantarum lactobacillus VB165, rhamnosus lactobacillus VB255 and animal bifidobacterium lactis subspecies VB301, but also include other strains with similar functions and characteristics within their species, which have similar effects in regulating intestinal flora, improving intestinal barrier function, reducing inflammatory response and controlling blood sugar levels. Therefore, the probiotic composition of the present invention is not limited to the above-mentioned specific strains, but can also include other strains within the same genus or species. As long as these strains can achieve the technical effects described in the present invention, they should all be considered as the scope of protection of the present invention.

[0032] It should be noted that the plantarum Lactobacillus VB165 involved in this application has a deposit number of CGMCC No. 25839, which has been disclosed in the application document with application number PCT / CN2024 / 107872, and the disclosed content includes information such as the source and function of the strain; the rhamnosus Lactobacillus VB255 involved in this application has a deposit number of CGMCC No. 26968, which has been disclosed in the application document with application number 202311380692.2, and the disclosed content includes information such as the source and function of the strain; the animal Bifidobacterium lactis subspecies VB301 involved in this application has a deposit number of CGMCC No. 27998, which has been disclosed in the application document with application number PCT / CN2024 / 118617, and the disclosed content includes information such as the source and function of the strain.

[0033] According to an embodiment of the present invention, the composition comprises the Lactobacillus plantarum and the Lactobacillus rhamnosus; the ratio of the viable counts of the Lactobacillus plantarum and the Lactobacillus rhamnosus is (1-5):(1-5). Thus, by adjusting the viable counts of Lactobacillus plantarum and Lactobacillus rhamnosus, the synergistic effect of the two strains is optimized, so that the probiotic composition can give full play to the advantages of the two strains, and further improve the ability of the prepared probiotic composition in regulating intestinal flora, controlling blood sugar levels and anti-inflammation; illustratively, the ratio of the viable counts of the Lactobacillus plantarum and the Lactobacillus rhamnosus can be 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 2:3, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:3, 4:5, 5:1, 5:2, 5:3, 5:4, etc., or can be a range consisting of any of the above values.

[0034] According to an embodiment of the present invention, the composition comprises the plant lactobacillus and the animal bifidobacterium lactis subspecies; the ratio of the number of viable bacteria of the plant lactobacillus and the animal bifidobacterium lactis subspecies is (1-5): (1-5). Thus, by adjusting the number of viable bacteria of the plant lactobacillus and the animal bifidobacterium lactis subspecies, the synergistic effect of the two strains is optimized, so that the probiotic composition can give full play to the advantages of the two strains, and further improve the ability of the prepared probiotic composition in regulating intestinal flora, controlling blood sugar levels and anti-inflammation; illustratively, the ratio of the number of viable bacteria of the plant lactobacillus and the animal bifidobacterium lactis subspecies can be 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 2:3, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:3, 4:5, 5:1, 5:2, 5:3, 5:4, etc., or can be a range composed of any of the above numerical values.

[0035] According to an embodiment of the present invention, the composition comprises the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis; the ratio of the number of live bacteria of the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis is (1-5): (1-5). Thus, by adjusting the number of live bacteria of the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis, the synergistic effect of the two strains is optimized, so that the probiotic composition can give full play to the advantages of the two strains, and further improve the ability of the prepared probiotic composition in regulating intestinal flora, controlling blood sugar levels and anti-inflammation; illustratively, the ratio of the number of live bacteria of the Lactobacillus rhamnosus and the Bifidobacterium animalis subspecies lactis can be 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 2:3, 2:5, 3:1, 3:2, 3:4, 3:5, 4:1, 4:3, 4:5, 5:1, 5:2, 5:3, 5:4, etc., or can be a range consisting of any of the above numerical values.

[0036] According to an embodiment of the present invention, the composition comprises the plant lactobacillus, the rhamnosus lactobacillus and the animal bifidobacterium lactis subspecies; the ratio of the number of live bacteria of the plant lactobacillus, the rhamnosus lactobacillus and the animal bifidobacterium lactis subspecies is (1-5): (1-5): (1-5). Thus, by adjusting the number of live bacteria of the plant lactobacillus, the rhamnosus lactobacillus and the animal bifidobacterium lactis subspecies, the synergistic effect of the three strains is optimized, so that the probiotic composition can give full play to the advantages of the three strains, and further improve the ability of the prepared probiotic composition in regulating intestinal flora, controlling blood sugar levels and anti-inflammation; illustratively, the ratio of the number of live bacteria of the plant lactobacillus, the rhamnosus lactobacillus and the animal bifidobacterium lactis subspecies can be 1:1:1, 1:2:1, 1:3:1, 1:4:1, 1:5:1, 2:1:1 , 2:3:1, 2:5:1, 3:1:1, 3:2:1, 3:4:1, 3:5:1, 4:1:1, 4:3:1, 4:5:1, 5:1:1, 5:2:1, 5:3:1, 5:4:1, 1:1:2, 1:2:2, 1:3:2, 1:4:2, 1:5:2, 2:1:2, 2:3:2, 2:5:2, 3:1:2, 3:2:2, 3:4:2, 3:5:2, 4:1:2, 4:3:2, 4:5:2, 5:1:2, 5:2:2, 5:3 :2, 5:4:2, 1:1:3, 1:2:3, 1:3:3, 1:4:3, 1:5:3, 2:1:3, 2:3:3, 2:5:3, 3:1:3, 3:2:3, 3:4:3, 3:5:3, 4:1:3, 4:3:3, 4:5:3, 5:1:3, 5:2:3, 5:3:3, 5:4:3, 1:1:4, 1:2:4, 1:3:4, 1:4:4, 1:5:4, 2:1:4, 2:3:4, 2:5:4, 3:1:4, 3:2:4, 3 :4:4, 3:5:4, 4:1:4, 4:3:4, 4:5:4, 5:1:4, 5:2:4, 5:3:4, 5:4:4, 1:1:5, 1:2:5, 1:3:5, 1:4:5, 1:5:5, 2:1:5, 2:3:5, 2:5:5, 3:1:5, 3:2:5, 3:4:5, 3:5:5, 4:1:5, 4:3:5, 4:5:5, 5:1:5, 5:2:5, 5:3:5, 5:4:5, etc., or it can be a range consisting of any of the above values.

[0037] Compound bacterial agent

[0038] The present invention provides a composite bacterial agent. According to an embodiment of the present invention, the active ingredient of the composite bacterial agent is the probiotic composition described in the first aspect. The composite bacterial agent according to an embodiment of the present invention, by incorporating the aforementioned probiotic composition into the composite bacterial agent, makes the probiotic composition easier to store, transport, and use, further improving its convenience and practicality, further expanding its application range, and facilitating use in different scenarios.

[0039] In some specific embodiments, the composite bacterial agent includes liquid bacterial agents and solid bacterial agents. Exemplarily, when the composite bacterial agent is in the form of a solid bacterial agent, it includes but is not limited to freeze-dried powder.

[0040] It should be noted that, in the composite bacterial agent of the present invention, Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis may exist in the form of live bacteria and / or non-live bacteria.

[0041] As used herein, "live bacterial form" refers to the form of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis that has the ability to metabolize, reproduce or replicate.

[0042] For example, the living bacteria may be immobilized bacteria. Herein, "immobilized bacteria" refers to living bacteria fixed on a carrier, which can carry out life activities such as growth, development, reproduction, inheritance and metabolism within a certain spatial range.

[0043] Herein, "non-viable bacterial form" refers to the existence form of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subspecies lactis that do not have the ability to metabolize, reproduce and replicate, including but not limited to dry bacteria. Exemplarily, the microbial agent is a freeze-dried powder.

[0044] In some specific embodiments, the Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subspecies lactis are in the form of live bacteria, dried bacteria, immobilized bacteria or any other forms.

[0045] In some specific embodiments, the dried bacteria are obtained by freeze-drying Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis.

[0046] Application in the preparation of food, health products or medicines

[0047] The present invention proposes the use of the aforementioned probiotic composition or composite bacterial agent in the preparation of food, health products, or pharmaceuticals. According to embodiments of the present invention, the pharmaceutical has at least one of the following uses: preventing, alleviating, assisting in the treatment, and / or treating diabetes; controlling blood sugar levels; and providing anti-inflammatory benefits. According to embodiments of the present invention, by applying the aforementioned probiotic composition to food, health products, or pharmaceuticals, particularly pharmaceuticals, a safer and more effective treatment option is provided for diabetic patients.

[0048] Application in the preparation of products for preventing, alleviating, assisting in the treatment or treating diabetes

[0049] The present invention proposes the use of the aforementioned probiotic composition or the aforementioned composite bacterial agent in the preparation of a product for preventing, alleviating, assisting in the treatment or curing diabetes.

[0050] It will be understood by those skilled in the art that the features and advantages described above for the probiotic composition or compound bacterial agent are also applicable to this application and will not be repeated here.

[0051] Below, the scheme of the present invention will be explained in conjunction with embodiment.It will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.In the embodiment, if specific technology or conditions are not indicated, the technology or conditions described in the literature in this area or the product instructions are used.The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0052] Example 1: Effect of probiotic composition on diabetic mouse model

[0053] 1. Adaptive breeding

[0054] Ninety healthy male SPF Kunming mice aged 8-9 weeks (purchased from Hangzhou Normal University School of Medicine) were selected and housed at a temperature of 20-26°C and a humidity of 40%-70%. The mice were placed in a light-dark cycle with 12-h alternation and had free access to food. The mice were adaptively housed for 7 days.

[0055] 2. Preparation of Probiotic Compositions I to V

[0056] Probiotic composition I: Lactobacillus plantarum VB165 and Lactobacillus rhamnosus VB255, the ratio of the number of viable bacteria is 1:1

[0057] Probiotic composition II: Lactobacillus plantarum VB165 and Bifidobacterium animalis subsp. lactis VB301, the ratio of the number of viable bacteria is 1:1

[0058] Probiotic composition III: Lactobacillus rhamnosus VB255 and Bifidobacterium animalis subsp. lactis VB301, the ratio of the number of viable bacteria of the two bacteria is 1:1

[0059] Probiotic composition IV: Lactobacillus plantarum VB165, Lactobacillus rhamnosus VB255 and Bifidobacterium animalis subsp. lactis VB301, the ratio of the number of viable bacteria is 1:1:1

[0060] Probiotic composition V: Lactobacillus rhamnosus B81450 (purchased from Ningbo Mingzhou Biotechnology Co., Ltd.), Lactobacillus plantarum VB165 and Bifidobacterium animalis subsp. lactis VB301, the ratio of the viable count of the three bacteria is 1:1:1

[0061] 3. Grouping of experimental mice

[0062] All mice were fasted for 16 h and blood was collected from the tip of their tails. The fasting blood glucose of each mouse was measured using a home blood glucose meter. The mice with fasting blood glucose higher than 7.1 mmol / L were eliminated. Finally, 70 experimental mice (body weight 20±3 g, fasting blood glucose ≤7.1 mmol / L) were obtained and randomly divided into 7 groups, with 10 mice in each group, namely control group (Ctrl), model group (T2DM), probiotic VB165+VB255 group (probiotic composition I), probiotic VB165+VB301 group (probiotic composition II), probiotic VB255+VB301 group (probiotic composition III), probiotic VB165+VB255+VB301 group (probiotic composition IV), and probiotic B81450+VB165+VB301 group (probiotic composition V).

[0063] 4. Construction of diabetic mouse model

[0064] (1) Construction of diabetic mouse models in the model group and probiotic intervention groups

[0065] A diabetic model was established by inducing streptozotocin (for specific methods, please refer to the existing technology: S. Liu, L. Ma, X. Ren, et al, A new mouse model of type 2 diabetes mellitus establishedthrough combination of high-fat diet, streptozotocin and glucocorticoid, LifeSci 286 (2021) 120062.). To construct a high-fat diet / STZ diabetes model, the experimental mice were fed a high-fat diet for 4 weeks. After 4 weeks, they were fasted for 6 hours and each experimental mouse was intraperitoneally injected with a single dose of STZ at a dose of 50 mg / kg (freshly dissolved in sodium citrate buffer, pH=4.5) for 5 consecutive days; fasting blood glucose was measured one week later, and a fasting blood glucose value greater than 11.1 mmol / L was considered to be a diabetic mouse model; the experimental mice maintained a high-fat diet until the end of the study.

[0066] (2) Control group experimental mice

[0067] The experimental mice in the control group continued to be fed with ordinary feed for 4 weeks. After 4 weeks, they were fasted for 6 hours and injected intraperitoneally with an equal amount of (50 mg / kg) sodium citrate buffer (pH = 4.5) for 5 consecutive days. In addition to the above diet and injection treatments, other conditions (such as housing environment, temperature, humidity, etc.) remained consistent with those of the model group and each probiotic intervention group.

[0068] 5. Oral gavage intervention

[0069] All experimental mice were given oral gavage intervention treatment. The control group and the model group were given 400 μL of deionized water by oral gavage every day; the probiotic VB165+VB255 group was given probiotic composition I (total viable bacteria concentration = 1×10 9 CFU, gavage volume 400 μL); probiotics VB165+VB301 group was gavaged daily with probiotic composition II suspended in deionized water (total viable bacteria concentration = 1×10 9 CFU, oral gavage volume 400 μL); probiotics VB255+VB301 group was gavaged daily with probiotic composition III suspended in deionized water (total viable bacteria concentration = 1×10 9 CFU, oral gavage volume 400 μL); probiotics VB165+VB255+VB301 group was gavaged daily with probiotic composition IV suspended in deionized water (total viable bacteria concentration = 1×10 9CFU, oral gavage volume 400 μL); probiotics B81450+VB165+VB301 group was gavaged daily with probiotic composition V suspended in deionized water (total viable bacteria concentration = 1×10 9 CFU, gavage volume was 400 μL).

[0070] 6. Fasting blood glucose and glucose tolerance test in mice

[0071] (1) Fasting blood glucose measurement

[0072] At week 0 before and week 8 after the intervention with probiotic compositions I to V, each experimental mouse was fasted for 6 h, its tail was disinfected with an alcohol cotton ball, and blood was collected from the tip of the tail. The fasting blood glucose of each experimental mouse was measured using a home blood glucose meter.

[0073] The fasting blood glucose test results of each experimental mouse before and after the intervention of probiotic composition I~V are shown in Figure 1 .

[0074] The results showed that compared with the control group, the fasting blood glucose values ​​of the experimental mice in the model group increased significantly, that is, the experimental mice in the model group showed hyperglycemia; but after intervention with probiotic compositions I to V, the blood glucose values ​​of the diabetic model mice decreased, among which the VB165+VB255+VB301 group (probiotic composition IV) had the best blood glucose intervention effect.

[0075] (2) Glucose tolerance test

[0076] After the eighth week of intervention with probiotic compositions I to V, the mice were fasted for 8 h and subjected to an oral glucose tolerance test with 2 g / kg glucose (20% solution). Tail tip blood samples were collected from the mice at 0, 30, 60, 90, and 120 min after glucose administration. Blood glucose levels were measured using a home blood glucose meter, and glucose concentration curves were plotted.

[0077] The results of glucose tolerance test of each experimental mouse after intervention with probiotic composition I to V are shown in Figure 2 .

[0078] The results showed that compared with the control group, the blood glucose levels of the experimental mice in the model group were significantly increased from 0 to 120 min; compared with the model group, the blood glucose levels of the diabetic model mice decreased to varying degrees at 60 min, 90 min and 120 min after the intervention of probiotic compositions I to V, among which the VB165+VB255+VB301 group (probiotic composition IV) had the best blood glucose intervention effect.

[0079] 7. Detection of inflammatory indicators in mice

[0080] At the end of the experiment, blood was collected from the mouse eyeballs and centrifuged at 3500 rpm for 10 min to obtain plasma. The plasma was diluted 10-fold and the concentrations of inflammatory factors IL-6 and IL-1β in mouse plasma were detected according to the instructions of the mouse IL-6 ELISA kit (purchased from Shenzhen Dakoway Biotechnology Co., Ltd.) and the mouse IL-1β ELISA kit (purchased from Shenzhen Dakoway Biotechnology Co., Ltd.).

[0081] The results of inflammatory factor detection in each experimental mouse after intervention with probiotic composition I~V are shown in Figure 3 .

[0082] The results showed that compared with the control group, the inflammatory factors IL-6 and IL-1β in the plasma of the experimental mice in the model group were significantly increased; compared with the model group, after the intervention of probiotic compositions I~V, the inflammatory factors IL-6 and IL-1β in the plasma of the diabetic model mice were significantly reduced, among which the VB165+VB255+VB301 group (probiotic composition IV) had the best anti-inflammatory effect.

[0083] The above results show that intervention with probiotic compositions I to V can reduce systemic inflammatory response and control blood sugar levels in diabetic mice, among which probiotic composition IV has the best effect in reducing inflammatory response and controlling blood sugar levels.

[0084] 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.

[0085] 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. Use of a probiotic composition or a composite bacterial agent in the preparation of medicines or health products, characterized in that: The drug has at least one of the following uses: Prevent, alleviate, assist in the treatment and / or cure of diabetes; Control blood sugar levels; The health care product has the following uses: Control blood sugar levels; Wherein, the probiotic composition or compound bacterial agent is composed of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis; The plant lactobacillus is plant lactobacillus ( Lactobacillus plantarum )VB165, whose deposit number is CGMCC No.25839; The Lactobacillus rhamnosus is Lactobacillus rhamnosus ( Lactobacillus rhamnosus )VB255, whose deposit number is CGMCC No.26968; The animal Bifidobacterium lactis subspecies is animal Bifidobacterium lactis subspecies ( Bifidobacterium animalis subsp. lactis ) VB301, whose deposit number is CGMCC No.27998; The ratio of the number of viable bacteria of the plant lactobacillus, the rhamnosus lactobacillus and the animal Bifidobacterium lactis subspecies is (1-5): (1-5): (1-5).

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