Probiotic composition for regulating blood glucose metabolism and application thereof

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a probiotic composition for regulating blood glucose metabolism and application thereof, and the probiotic composition comprises the following components: two or three of lactobacillus plantarum, lactobacillus rhamnosus and bifidobacterium animalis subsp. Lactis. The invention provides a probiotic composition, which comprises two or three of lactobacillus plantarum, lactobacillus rhamnosus and bifidobacterium animalis subsp. Lactis, and further optimizes the synergistic effect of strains by adjusting the variety collocation of the strains and the proportion of viable count, so that the probiotic composition can be used for preparing probiotics. The probiotic composition has better effects in the aspects of preventing and treating diabetes mellitus, controlling blood sugar level, regulating intestinal flora, resisting inflammation and the like, in addition, the probiotic composition can also be applied to prevention, relieving, adjuvant therapy or treatment of diabetes mellitus, and a more effective and safer solution is provided for management of diabetes mellitus.

Description

Technical Field

[0001] The present invention belongs to the field of probiotics, and particularly relates to a probiotic composition for regulating blood glucose metabolism and its application. Background Art

[0002] Diabetes is a metabolic disease characterized by chronic hyperglycemia. Long-term hyperglycemia causes chronic damage or dysfunction of various tissues of the body, especially the eyes, kidneys, heart, blood vessels, and nerves. As a global public health problem, in recent years, the incidence rate has shown a significant upward trend, bringing a heavy economic burden to patients and their families. Currently, the prevention and treatment of diabetes mainly rely on drug therapy, diet control, and exercise therapy. However, these methods still have many deficiencies. Drug therapy may cause side effects such as hypoglycemia and gastrointestinal reactions; the effects of diet control and exercise therapy vary from person to person and are difficult to adhere to in the long term.

[0003] Related studies have found that the gut microbiota is closely related to diabetes, and regulating the gut microbiota has become a new direction in diabetes treatment research. Currently, some probiotic strains have been found to have a positive effect on the prevention and treatment of diabetes, but there are problems such as a single formula and limited effects in existing probiotic products.

[0004] Therefore, developing a probiotic composition that can effectively prevent or treat diabetes is of great significance for improving the prognosis of diabetes patients and reducing the medical costs of diabetes 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 some extent. For this purpose, the present invention provides a probiotic composition for regulating blood glucose metabolism and its application. The present invention provides a probiotic composition, which comprises two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis; wherein, the Lactobacillus plantarum is Lactobacillus plantarum VB165, and its deposit number is CGMCC No. 25839; the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, and its deposit number is CGMCC No. 26968; the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis VB301, and its deposit number is CGMCC No. 27998. By adjusting the type combination and viable count ratio 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 glucose levels, regulating the gut microbiota, and anti-inflammation. In addition, the probiotic composition can also be applied to prevent, relieve, adjuvantly treat, or treat diabetes, providing a more effective and safer solution for diabetes management.

[0006] In the first aspect of the present invention, the present invention provides 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 subsp. lactis; wherein, the Lactobacillus plantarum is Lactobacillus plantarum ( Lactobacillus plantarum ), VB165, with a deposit number of CGMCC No. 25839, deposit date: September 28, 2022, depositary institution: China General Microbiological Culture Collection Center; address of the depositary institution: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the Lactobacillus rhamnosus is Lactobacillus rhamnosus ( Lactobacillus rhamnosus ), VB255, with a deposit number of CGMCC No. 26968; deposit date: March 31, 2023, depositary institution: China General Microbiological Culture Collection Center; address of the depositary institution: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis ( Bifidobacterium animalis subsp. lactis ), VB301, with a deposit number of CGMCC No. 27998; deposit date: July 24, 2023, depositary institution: China General Microbiological Culture Collection Center; address of the depositary institution: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The probiotic composition according to the embodiment of the present invention, which contains two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis, can more effectively regulate the intestinal flora, enhance the intestinal barrier function, and reduce the inflammatory response under the synergistic action of these strains, thus showing significant effects in the prevention and treatment of diabetes, blood glucose level control, and anti-inflammation, and having a wide application prospect.

[0007] According to an embodiment of the present invention, the above probiotic composition may further have the following additional technical features: According to an embodiment of the present invention, it contains the Lactobacillus plantarum and the Lactobacillus rhamnosus; the viable count ratio of the Lactobacillus plantarum to the Lactobacillus rhamnosus is (1~5):(1~5).

[0008] According to an embodiment of the present invention, it contains the Lactobacillus plantarum and the Bifidobacterium animalis subsp. lactis; the viable count ratio of the Lactobacillus plantarum to the Bifidobacterium animalis subsp. lactis is (1~5):(1~5).

[0009] According to an embodiment of the present invention, it contains the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis; the viable count ratio of the Lactobacillus rhamnosus to the Bifidobacterium animalis subsp. lactis is (1~5):(1~5).

[0010] According to an embodiment of the present invention, it includes the Lactobacillus plantarum, the Lactobacillus rhamnosus, and the Bifidobacterium animalis subsp. lactis; the viable count ratio 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 compound bacterium agent. According to an embodiment of the present invention, the active ingredient of the compound bacterium agent is the probiotic composition described in the first aspect. By applying the aforementioned probiotic composition to the compound bacterium agent, the probiotic composition is more convenient for storage, transportation and use, further improving its convenience and practicality in use, further expanding its application scope, and facilitating its use in different scenarios.

[0012] In a third aspect of the present invention, the present invention provides the application of the probiotic composition described in the first aspect or the compound bacterium agent described in the second aspect in the preparation of food, health products or drugs. According to an embodiment of the present invention, the drug has at least one of the following uses: preventing, alleviating, adjuvantly treating and / or treating diabetes; controlling blood glucose levels; anti-inflammatory. Through the application of the aforementioned probiotic composition to food, health products or drugs, when it is applied to drugs, it provides a safer and more effective treatment option for diabetic patients.

[0013] In a fourth aspect of the present invention, the present invention provides the application of the probiotic composition described in the first aspect or the compound bacterium agent described in the second aspect in the preparation of products for preventing, alleviating, adjuvantly treating or treating diabetes.

[0014] Those skilled in the art can understand that the features and advantages described above for the probiotic composition or the compound bacterium agent also apply to this application, and will not be repeated here.

[0015] Additional aspects and advantages of the present invention will be given in part in the following description, become apparent in part from the following description, or be learned through the practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The above and / or additional aspects and advantages of the present invention will become apparent and be readily understood from the description of the embodiments in conjunction with the following drawings, wherein: Figure 1 It is a graph showing the fasting blood glucose measurement results of each experimental mouse before and after the intervention of Probiotic Combinations I-V in Example 1 of the present invention. Among them, NS indicates that there is no significant difference in the data results of this part, * is P <0.05, ** is P <0.01, *** is P <0.001; Figure 2This is the graph showing the results of the glucose tolerance test for each experimental mouse after the intervention of probiotic compositions I-V in Example 1 of the present invention; Figure 3 This is the graph showing the detection results of inflammatory factors for each experimental mouse after the intervention of probiotic compositions I-V in Example 1 of the present invention. Among them, A is the graph showing the detection results of inflammatory factor IL-6 for each experimental mouse after the intervention of probiotic compositions I-V, and B is the graph showing the detection results of inflammatory factor IL-1β for each experimental mouse after the intervention of probiotic compositions I-V. Among them, * is P <0.05, ** is P <0.01, *** is P <0.001. Detailed implementation manners

[0017] The embodiments of the present invention will be described in detail below. The following described embodiments are exemplary and are only used to explain the present invention, and should not be construed as a limitation to the present invention.

[0018] It should be noted that the terms "first" and "second" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include one or more of such features. Further, in the description of the present invention, unless otherwise stated, the meaning of "a plurality" is two or more.

[0019] In the ranges disclosed herein, the endpoints and any value are not limited to the exact range or value. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and a single point value, and between single point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0020] In this article, the term "comprising" or "including" is an open expression, that is, it includes the content specified by the present invention, but does not exclude other aspects of the content.

[0021] In this article, the terms "optionally", "optional" or "option" generally mean that the subsequent events or conditions may or may not occur, and this description includes the cases where such events or conditions occur, as well as the cases where such events or conditions do not occur.

[0022] Probiotic composition The present invention provides 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 subsp. lactis; wherein, the Lactobacillus plantarum is Lactobacillus plantarum VB165, and its deposit number is CGMCC No. 25839; the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, and its deposit number is CGMCC No. 26968; the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis VB301, and its deposit number is CGMCC No. 27998. The probiotic composition according to the embodiment of the present invention comprises two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis. Under the synergistic action of these strains, it can more effectively regulate the intestinal flora, enhance the intestinal barrier function, reduce the inflammatory response, and thus show significant effects in the prevention and treatment of diabetes, blood glucose level control, and anti-inflammation, with broad application prospects.

[0023] According to an embodiment of the present invention, the Lactobacillus plantarum is Lactobacillus plantarum VB165, and its deposit number is CGMCC No. 25839. Thus, by optimizing the strain source of Lactobacillus plantarum, it provides an important basis for the subsequent standardized production and quality control of the probiotic composition. In addition, this strain itself can effectively inhibit the activity of α-glucosidase, and at the same time, it also has the ability to degrade bile salts, inhibit the activity of major intestinal pathogens, and has good tolerance to gastric acid and choline.

[0024] According to an embodiment of the present invention, the Lactobacillus rhamnosus is Lactobacillus rhamnosus VB255, and its deposit number is CGMCC No. 26968. Thus, by optimizing the strain source of Lactobacillus rhamnosus, it provides an important basis for the subsequent standardized production and quality control of the probiotic composition. In addition, this strain itself has good stability, can stably exist in pancreatic juice without affecting the intestinal microecological environment, and has a good inhibitory effect on Candida albicans and / or Shigella.

[0025] According to an embodiment of the present invention, the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis VB301, and its deposit number is CGMCC No. 27998. Thus, by optimizing the strain source of Bifidobacterium animalis subsp. lactis, it provides an important basis for the subsequent standardized production and quality control of the probiotic composition. In addition, this strain itself has a significant bile salt-lowering ability, and has strong intestinal tolerance and antibacterial ability.

[0026] It should be noted that the Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis in the probiotic composition of the present invention do not only include Lactobacillus plantarum VB165, Lactobacillus rhamnosus VB255, and Bifidobacterium animalis subsp. lactis VB301, but also include other strains within their species that have similar functions and characteristics. These strains have similar effects in regulating the intestinal flora, improving the intestinal barrier function, reducing the inflammatory response, and controlling blood glucose levels. Therefore, the probiotic composition described in the present invention is not limited to the above specific strains, but may 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 regarded as the protection scope of the present invention.

[0027] It should be noted that Lactobacillus plantarum VB165 involved in the present application, with the deposit number of CGMCC No. 25839, has been disclosed in the application document with the application number of PCT / CN2024 / 107872, and the disclosure content includes information such as the source and function of the strain; Lactobacillus rhamnosus VB255 involved in the present application, with the deposit number of CGMCC No. 26968, has been disclosed in the application document with the application number of 202311380692.2, and the disclosure content includes information such as the source and function of the strain; Bifidobacterium animalis subsp. lactis VB301 involved in the present application, with the deposit number of CGMCC No. 27998, has been disclosed in the application document with the application number of PCT / CN2024 / 118617, and the disclosure content includes information such as the source and function of the strain.

[0028] According to the embodiments of the present invention, it includes the Lactobacillus plantarum and the Lactobacillus rhamnosus; the ratio of the viable cell count of the Lactobacillus plantarum to the Lactobacillus rhamnosus is (1 - 5):(1 - 5). Thus, by adjusting the viable cell count of the Lactobacillus plantarum and the Lactobacillus rhamnosus and optimizing the synergistic effect of these two strains, 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 the intestinal flora, controlling blood glucose levels, and anti - inflammation; exemplarily, the ratio of the viable cell count of the Lactobacillus plantarum to 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 composed of any of the above values.

[0029] According to an embodiment of the present invention, it contains the Lactobacillus plantarum and the Bifidobacterium animalis subsp. lactis; the ratio of the viable cell count of the Lactobacillus plantarum to that of the Bifidobacterium animalis subsp. lactis is (1-5):(1-5). Thus, by adjusting the viable cell counts of the Lactobacillus plantarum and the Bifidobacterium animalis subsp. lactis, the synergistic effect of these two strains is optimized, enabling the probiotic composition to fully exert the advantages of the two strains and further enhancing the ability of the prepared probiotic composition in regulating the intestinal flora, controlling blood glucose levels, and anti-inflammatory aspects; exemplarily, the ratio of the viable cell count of the Lactobacillus plantarum to that of the Bifidobacterium animalis subsp. 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 composed of any of the above values.

[0030] According to an embodiment of the present invention, it contains the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis; the ratio of the viable cell count of the Lactobacillus rhamnosus to that of the Bifidobacterium animalis subsp. lactis is (1-5):(1-5). Thus, by adjusting the viable cell counts of the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis, the synergistic effect of these two strains is optimized, enabling the probiotic composition to fully exert the advantages of the two strains and further enhancing the ability of the prepared probiotic composition in regulating the intestinal flora, controlling blood glucose levels, and anti-inflammatory aspects; exemplarily, the ratio of the viable cell count of the Lactobacillus rhamnosus to that of the Bifidobacterium animalis subsp. 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 composed of any of the above values.

[0031] According to an embodiment of the present invention, it includes the Lactobacillus plantarum, the Lactobacillus rhamnosus, and the Bifidobacterium animalis subsp. lactis; the viable count ratio of the Lactobacillus plantarum, the Lactobacillus rhamnosus, and the Bifidobacterium animalis subsp. lactis is (1-5):(1-5):(1-5). Thus, by adjusting the viable counts of the Lactobacillus plantarum, the Lactobacillus rhamnosus, and the Bifidobacterium animalis subsp. lactis, the synergistic effect of these three strains is optimized, enabling the probiotic composition to fully exert the advantages of the three strains and further enhancing the ability of the prepared probiotic composition in regulating the intestinal flora, controlling blood glucose levels, and anti-inflammation; exemplarily, the viable count ratio of the Lactobacillus plantarum, the Lactobacillus rhamnosus, and the Bifidobacterium animalis subsp. lactis 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 can be a range composed of any of the above numerical values.

[0032] Compound bacterial agent The present invention provides a composite microbial agent. According to an embodiment of the present invention, the active ingredient of the composite microbial agent is the probiotic composition described in the first aspect. The composite microbial agent according to the embodiment of the present invention makes the probiotic composition easier to store, transport and use by applying the aforementioned probiotic composition to the composite microbial agent, further improving its convenience and practicality in use, further expanding its application scope, and facilitating its use in different scenarios.

[0033] In some specific embodiments, the forms of the composite microbial agent include two types: liquid microbial agent and solid microbial agent. Exemplarily, when the form of the composite microbial agent is a solid microbial agent, it includes but is not limited to freeze-dried powder, etc.

[0034] It should be noted that in the composite microbial agent of the present invention, Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis can exist in the form of viable bacteria and / or non-viable bacteria.

[0035] In this article, the "form of viable bacteria" refers to the existence form of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis with the ability of metabolism, reproduction or replication.

[0036] Exemplarily, the viable bacteria can be immobilized bacteria. In this article, "immobilized bacteria" refers to live bacteria immobilized on a carrier, which can carry out life activities such as growth, development, reproduction, inheritance and metabolism within a certain spatial range.

[0037] In this article, the "form of non-viable bacteria" refers to the existence form of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis without the ability of metabolism, reproduction and replication, including but not limited to dried bacterial cells. Exemplarily, the microbial agent is freeze-dried powder.

[0038] In some specific embodiments, Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis exist in the form of viable bacteria, dried bacterial cells, immobilized bacteria or any other form.

[0039] In some specific embodiments, the dried bacterial cells are obtained by freeze-drying Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis.

[0040] Application in the preparation of food, health products or drugs The present invention provides the application of the aforementioned probiotic composition or the aforementioned composite microbial agent in the preparation of food, health products or drugs. According to an embodiment of the present invention, the drug has at least one of the following uses: preventing, alleviating, adjuvantly treating and / or treating diabetes; controlling blood glucose level; anti-inflammatory. According to the application of the embodiment of the present invention, by applying the aforementioned probiotic composition to food, health products or drugs, and applying it to drugs, a safer and more effective treatment option is provided for diabetic patients.

[0041] Use in the preparation of a product for preventing, alleviating, adjuvantly treating or treating diabetes The present invention provides the use of the aforementioned probiotic composition or the aforementioned complex bacterial agent in the preparation of a product for preventing, alleviating, adjuvantly treating or treating diabetes.

[0042] Those skilled in the art can understand that the features and advantages described above for the probiotic composition or the complex bacterial agent also apply to this use and will not be elaborated herein.

[0043] The solution of the present invention will be explained below in conjunction with examples. Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. For those not specified in the examples regarding specific techniques or conditions, they shall be carried out according to the techniques or conditions described in the literature in this field or according to the product specifications. For those reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0044] Example 1: Influence of the probiotic composition on a diabetic mouse model 1. Adaptive feeding Ninety 8- to 9-week-old healthy male SPF-grade Kunming mice (purchased from the School of Medicine, Hangzhou Normal University) were selected. The feeding temperature was set at 20-26 °C, the humidity was set at 40%-70%, and the light and dark alternated every 12 h. All mice were allowed to eat and drink freely and were adaptively fed for 7 days.

[0045] 2. Preparation of probiotic compositions I-V Probiotic composition I: Lactobacillus plantarum VB165 and Lactobacillus rhamnosus VB255, the viable cell number ratio of the two bacteria = 1:1 Probiotic composition II: Lactobacillus plantarum VB165 and Bifidobacterium animalis subsp. lactis VB301, the viable cell number ratio of the two bacteria = 1:1 Probiotic composition III: Lactobacillus rhamnosus VB255 and Bifidobacterium animalis subsp. lactis VB301, the viable cell number ratio of the two bacteria = 1:1 Probiotic composition IV: Lactobacillus plantarum VB165, Lactobacillus rhamnosus VB255 and Bifidobacterium animalis subsp. lactis VB301, the viable cell number ratio of the three bacteria = 1:1:1 Probiotic composition V: Lactobacillus rhamnosus B81450 (purchased from Ningbo Mingzhou Biotechnology Co., Ltd.), Lactobacillus plantarum VB165 and Bifidobacterium animalis subsp. lactis VB301, the viable cell number ratio of the three bacteria = 1:1:1 3. Grouping of experimental mice After fasting all mice for 16 h, tail tip blood was collected, and the fasting blood glucose of each mouse was measured using a household blood glucose meter. Mice with fasting blood glucose higher than 7.1 mmol / L were excluded. Finally, 70 experimental mice (body weight 20 ± 3 g, fasting blood glucose ≤ 7.1 mmol / L) were randomly divided into 7 groups, with 10 mice in each group, namely the control group (Ctrl), the model group (T2DM), the probiotic VB165 + VB255 group (probiotic composition I), the probiotic VB165 + VB301 group (probiotic composition II), the probiotic VB255 + VB301 group (probiotic composition III), the probiotic VB165 + VB255 + VB301 group (probiotic composition IV), and the probiotic B81450 + VB165 + VB301 group (probiotic composition V).

[0046] 4. Construction of diabetic mouse model (1)Construction of diabetic mouse models in the model group and each probiotic intervention group A diabetic model was induced by streptozotocin (for the specific method, refer to the prior art: S. Liu, L. Ma, X. Ren, et al, A new mouse model of type 2 diabetes mellitus established through combination of high - fat diet, streptozotocin and glucocorticoid, Life Sci 286 (2021) 120062.). To construct a high - fat diet / STZ diabetic model, experimental mice were fed a high - fat diet for 4 weeks. After 4 weeks, they were fasted for 6 h, and a single dose of STZ at a dose of 50 mg / kg (freshly dissolved in sodium citrate buffer, pH = 4.5) was intraperitoneally injected into each experimental mouse for 5 consecutive days; one week later, the fasting blood glucose was measured, and a fasting blood glucose value greater than 11.1 mmol / L was considered a diabetic mouse model; the experimental mice maintained a high - fat diet until the end of the study.

[0047] (2)Control group experimental mice Control group experimental mice continued to be fed a normal diet for 4 weeks. After 4 weeks, they were fasted for 6 h, and an equal amount (50 mg / kg) of sodium citrate buffer (pH = 4.5) was intraperitoneally injected into each experimental mouse for 5 consecutive days; except for the above - mentioned diet and injection treatments, other conditions (such as the breeding environment, temperature, humidity, etc.) were kept the same as those in the model group and each probiotic intervention group.

[0048] 5. Gavage intervention treatment Each group of experimental mice was given intragastric intervention. The control group and the model group were given 400 μL of deionized water by gavage every day; the probiotic VB165+VB255 group was given the probiotic composition I suspended in deionized water by gavage every day (total viable concentration = 1×10 9 CFU, gavage volume was 400 μL); the probiotic VB165+VB301 group was given the probiotic composition II suspended in deionized water by gavage every day (total viable concentration = 1×10 9 CFU, gavage volume was 400 μL); the probiotic VB255+VB301 group was given the probiotic composition III suspended in deionized water by gavage every day (total viable concentration = 1×10 9 CFU, gavage volume was 400 μL); the probiotic VB165+VB255+VB301 group was given the probiotic composition IV suspended in deionized water by gavage every day (total viable concentration = 1×10 9 CFU, gavage volume was 400 μL); the probiotic B81450+VB165+VB301 group was given the probiotic composition V suspended in deionized water by gavage every day (total viable concentration = 1×10 9 CFU, gavage volume was 400 μL).

[0049] 6. Fasting blood glucose and glucose tolerance tests in mice (1)Determination of fasting blood glucose Before the intervention with probiotic compositions I-V at 0 week and after the 8th week of intervention, each experimental mouse was fasted for 6 h, the tail was disinfected with an alcohol cotton ball, and tail tip blood was taken. The fasting blood glucose of each experimental mouse was detected using a household blood glucose meter.

[0050] The results of the determination of fasting blood glucose of each experimental mouse before and after the intervention with probiotic compositions I-V are shown in Figure 1 .

[0051] The results showed that: compared with the control group, the fasting blood glucose value of the experimental mice in the model group increased significantly, that is, the experimental mice in the model group showed hyperglycemia; but after the intervention with probiotic compositions I-V, the blood glucose value of the diabetic model mice decreased. Among them, the VB165+VB255+VB301 group (probiotic composition IV) had the best effect on intervening blood glucose.

[0052] (2)Glucose tolerance test After the 8th week of intervention with probiotic compositions I-V, each experimental mouse was fasted for 8 h, and an oral glucose tolerance test was performed with 2 g / kg glucose (20% solution); and tail tip blood samples of each experimental mouse were collected at 0 min, 30 min, 60 min, 90 min, and 120 min after giving glucose, and the blood glucose value was detected using a household blood glucose meter, and a glucose concentration curve was plotted.

[0053] The determination results of the glucose tolerance test of each experimental mouse after the intervention of probiotic compositions I-V are shown in Figure 2 .

[0054] 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, at 60 min, 90 min and 120 min after the intervention of probiotic compositions I-V, the blood glucose levels of the diabetic model mice showed varying degrees of decrease. Among them, the VB165+VB255+VB301 group (probiotic composition IV) had the best effect on intervening blood glucose.

[0055] 7. Detection of inflammatory indexes in mice At the end of the experiment, blood was collected from the eyeballs of the mice and centrifuged at 3500 rpm for 10 min to obtain plasma; after diluting the plasma 10 times, the concentrations of inflammatory factors IL-6 and IL-1β in the plasma of the mice were detected according to the instructions of the mouse IL-6 ELISA kit (purchased from Shenzhen Dakewei Biotechnology Co., Ltd.) and the mouse IL-1β ELISA kit (purchased from Shenzhen Dakewei Biotechnology Co., Ltd.).

[0056] The detection results of the inflammatory factors of each experimental mouse after the intervention of probiotic compositions I-V are shown in Figure 3 .

[0057] 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 both 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 both significantly decreased. Among them, the VB165+VB255+VB301 group (probiotic composition IV) had the best anti-inflammatory effect.

[0058] The above results indicate that: the interventions of probiotic compositions I-V can all reduce the systemic inflammatory response and control the blood glucose level of diabetic mice. Among them, probiotic composition IV has the best effect in reducing the inflammatory response and controlling the blood glucose level.

[0059] In the description of this specification, the descriptions referring to terms such as "one embodiment", "some embodiments", "examples", "specific examples", or "some examples", etc., mean that the specific features, structures, materials, or characteristics described in connection 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, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0060] Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those of ordinary skill in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention.

Claims

1. A probiotic composition, characterized in that: The probiotic composition comprises the following components: two or three of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis; in, 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, its deposit number is CGMCC No.27998.

2. The probiotic composition according to claim 1, characterized in that Containing the Lactobacillus plantarum and the Lactobacillus rhamnosus; The ratio of the viable counts of the plant lactobacillus and the rhamnosus lactobacillus is (1-5): (1-5).

3. The probiotic composition according to claim 1, characterized in that comprising said Lactobacillus plantarum and said Bifidobacterium animalis subsp. lactis; The ratio of the number of live bacteria of the plant lactobacillus to that of the animal Bifidobacterium lactis subspecies is (1-5):(1-5).

4. The probiotic composition according to claim 1, characterized in that comprising the Lactobacillus rhamnosus and the Bifidobacterium animalis subsp. lactis; The ratio of the viable bacterial count of the Lactobacillus rhamnosus to the viable bacterial count of the Bifidobacterium animalis subspecies lactis is (1-5): (1-5).

5. The probiotic composition according to claim 1, characterized in that comprising the plant lactobacillus, the rhamnosus lactobacillus and the animal bifidobacterium lactis subsp. 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).

6. A composite bacterial agent, characterized in that: The active ingredient of the composite bacterial agent is the probiotic composition according to any one of claims 1 to 5.

7. Use of the probiotic composition according to any one of claims 1 to 5 or the composite bacterial agent according to claim 6 in the preparation of food, health products or medicines, characterized in that: The drug has at least one of the following uses: Prevention, alleviation, assistance and / or treatment of diabetes; Control blood sugar levels; Anti-inflammatory.

8. Use of the probiotic composition according to any one of claims 1 to 5 or the composite bacterial agent according to claim 6 in the preparation of a product for preventing, alleviating, assisting in the treatment or curing diabetes.

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