Probiotic composition with effects of losing weight and reducing blood sugar and application of probiotic composition

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a probiotic composition with the effects of losing weight and reducing blood sugar and application of the probiotic composition. The probiotic composition is prepared from the following raw materials: lactobacillus gasseri and lactobacillus reuteri, wherein the lactobacillus gasseri is a culture or a progeny culture of lactobacillus gasseri LG08. The lactobacillus reuteri is a culture or a progeny culture of the lactobacillus reuteri PRS-156 with the preservation number of CCTCC (China Center for Type Culture Collection) NO: M2023591 (China Center for Type Culture Collection). According to the probiotic composition containing lactobacillus gasseri and lactobacillus reuteri in a specific proportion and specific content, obesity and hyperglycemia diseases or symptoms can be obviously improved, and the probiotic composition has a good application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and particularly relates to a probiotic composition with weight loss and blood sugar lowering effects and its application. Background Art

[0002] In recent years, with the improvement of people's living standards, the change of lifestyle, the increase in high-calorie diet intake, and the decrease in the demand for physical activities in work and life, the imbalance between energy metabolism and intake of the body has occurred, resulting in an increase in overweight or obesity. Obesity should not be merely regarded as abnormal or excessive fat accumulation, and it has long become an important factor affecting people's health. The fundamental cause of obesity is the imbalance between energy intake and metabolism, which is closely related to various chronic diseases, such as cardiovascular and cerebrovascular diseases, non-alcoholic fatty liver, type II diabetes, etc.

[0003] Common weight loss drugs for the treatment of obesity currently include liraglutide, orlistat, sibutramine, rimonabant, etc. However, many weight loss drugs are restricted by the market or withdrawn from clinical applications because some of these drugs cannot achieve the expected effects or cause adverse reactions in patients. Therefore, there is an urgent need to develop an effective method or drug for treating obesity and related diseases with few side effects.

[0004] Research shows that probiotics represented by lactic acid bacteria have functions such as enhancing intestinal function, reducing cholesterol in serum, regulating lipid and glucose metabolism, inhibiting weight gain, and enhancing the immune system. Therefore, controlling obesity and its related metabolic diseases by ingesting probiotics has gradually become a research hotspot. While daily consuming a high-energy diet, it is of great significance to find probiotics with potential weight loss effects as dietary supplements to effectively inhibit the occurrence of obesity and its complications.

[0005] Chinese Patent No. 201910941360.4 discloses a weight loss composition and its preparation method. Among them, the raw materials of each portion of the weight loss composition include: 4 billion - 16 billion inactivated Akk bacteria, 2 billion - 10 billion natto bacteria, 1 billion - 6 billion Lactobacillus reuteri, and 5 g - 20 g of prebiotics by weight. This invention adopts multiple weight loss mechanisms to enhance the weight loss effect of the composition, overcoming the drawbacks of the single weight loss mechanism and poor weight loss effect of previous weight loss products; although the prior art has made certain explorations on probiotic weight loss and achieved certain effects, the strains involved are numerous and the preparation process is relatively complex, and further research and improvement are still needed. Summary of the Invention

[0006] Aiming at the deficiencies of the existing technology, the present invention provides a probiotic composition with weight loss and blood sugar lowering effects and its application.

[0007] To achieve the object of the present invention, the following technical solutions are adopted:

[0008] A probiotic composition with the effects of weight loss and blood sugar reduction, wherein the raw materials of the probiotic composition are Lactobacillus gasseri and Lactobacillus mucosae reuteri.

[0009] Preferably, the viable count ratio of Lactobacillus gasseri to Lactobacillus mucosae reuteri is 0.5 - 2:1.

[0010] Preferably, the Lactobacillus gasseri is a culture or subculture of Lactobacillus gasseri LG08.

[0011] Preferably, the Lactobacillus gasseri is derived from a culture or subculture of Lactobacillus gasseri LG08 of Jiangsu Microkang Biotechnology Co., Ltd., and can be cultured by existing methods.

[0012] Preferably, the Lactobacillus mucosae reuteri was deposited at the China Center for Type Culture Collection on April 23, 2023. The strain name is Lactobacillus mucosae reuteri PRS - 156, the Latin name is Limosilactobacillus reuteri PRS - 156, and the deposit number is CCTCC NO: M2023591. The deposit address is: Wuhan University, China. This strain has been disclosed in Chinese Invention Patent CN202410080900.5 and published in the patent gazette.

[0013] Preferably, the Lactobacillus mucosae reuteri is a culture or subculture of Lactobacillus mucosae reuteri PRS - 156.

[0014] The second object of the present invention is to provide the application of the above probiotic composition in the preparation of drugs for treating and / or preventing obesity or obesity complications.

[0015] The obesity includes, but is not limited to, genetic obesity and acquired obesity.

[0016] The acquired obesity includes, but is not limited to: obesity caused by a high - fat diet, obesity caused by drugs, obesity caused by endocrine disorders, obesity caused by lack of sleep, obesity caused by metabolic disorders, obesity caused by mental factors, or obesity caused by aging.

[0017] The obesity caused by mental factors includes, but is not limited to: obesity caused by vagus nerve excitation or obesity caused by sympathetic nerve excitation.

[0018] Preferably, the obesity complications are selected from one or more of hypertension, hyperlipidemia, hyperglycemia, fatty liver, gallstones, cholecystitis, hyperuricemia, osteoporosis, gout, coronary heart disease, myocardial infarction, cerebral infarction, and stroke;

[0019] Preferably, the obesity complication is selected from hyperglycemia.

[0020] Preferably, Lactobacillus gasseri and Limosilactobacillus reuteri can be in the form of bacterial powder or bacterial liquid when applied.

[0021] Preferably, the viable bacteria content in the bacterial liquid is 1×10 8 -1×10 10 CFU / mL; preferably 5×10 8 -5×10 10 CFU / mL; more preferably 1×10 9 CFU / mL.

[0022] The present invention also provides a preparation method of the aforementioned microbial composition.

[0023] The preparation method includes the cultivation of the above-mentioned Lactobacillus gasseri and Limosilactobacillus reuteri. The cultivation of Lactobacillus gasseri includes the following steps:

[0024] (1) Cultivation of Lactobacillus gasseri and activation of Limosilactobacillus reuteri strains;

[0025] (2) Cultivation of Lactobacillus gasseri and cultivation of Limosilactobacillus reuteri strains.

[0026] Preferably, the culture medium for the cultivation of Lactobacillus gasseri and the cultivation of Limosilactobacillus reuteri strains is MRS medium;

[0027] More preferably, the formulation of the MRS medium is as shown in Table 1 below:

[0028] Table 1

[0029] Component Content (g / L) Peptone 10.0 Beef extract 8.0 Yeast extract 4.0 Glucose 20.0 Dipotassium hydrogen phosphate 2.0 Ammonium hydrogen citrate 2.0 Sodium acetate 5.0 Magnesium sulfate 0.2 Manganese sulfate 0.04 Tween 80 1.0

[0030] Cultivation conditions: Under anaerobic conditions at a constant temperature of 37°C, cultivate in MRS medium for about 18 - 24h.

[0031] The third object of the present invention is to provide a bacterial agent, comprising a probiotic composition.

[0032] The fourth object of the present invention is to provide a drug for treating and / or preventing obesity or obesity complications, characterized in that the raw materials of the drug include the aforementioned probiotic composition or the bacterial agent.

[0033] The fifth object of the present invention is to provide the application of the aforementioned probiotic composition in the preparation of products that help control body fat and / or help maintain a healthy blood sugar level.

[0034] Compared with the prior art, the beneficial effects of the present invention:

[0035] The present invention can significantly improve obesity and hyperglycemia diseases or symptoms through a probiotic composition containing Lactobacillus gasseri and Limosilactobacillus reuteri with specific proportions and specific contents, and has good application prospects. Description of the Drawings

[0036] Figure 1 It shows the weight changes of mice in each group every 4 days from the 0th day of drug administration to the 32nd day of drug administration.

[0037] Figure 2 It shows the relative weight changes of mice in each group from the 0th day to the 32nd day of drug administration after successful modeling.

[0038] Figure 3 It shows the blood glucose of mice in each group.

[0039] Figure 4 It shows the Lee's index of mice in each group. Detailed Embodiments

[0040] The present invention will be further described below in conjunction with the detailed embodiments. The raw materials in the following examples are all commercially available conventional raw materials.

[0041] Among them, Lactobacillus gasseri is derived from the culture or progeny culture of Lactobacillus gasseri LG08 of Jiangsu Microkang Biotechnology Co., Ltd.

[0042] Limosilactobacillus reuteri was deposited at the China Center for Type Culture Collection on April 23, 2023. The strain name is Limosilactobacillus reuteri PRS-156, and the Latin name is Limosilactobacillus reuteri PRS-156. The deposit number is CCTCC NO: M2023591. The deposit address is: Wuhan University, China. This strain has been disclosed in Chinese invention patent CN202410080900.5 and the patent gazette.

[0043] Example 1

[0044] Culture of Lactobacillus gasseri LG08 and Limosilactobacillus reuteri PRS-156

[0045] (1) Strain activation: Lactobacillus gasseri LG08 and Limosilactobacillus reuteri PRS-156 used in the present invention were respectively inoculated into 50 mL of MRS medium and cultured in a shaker flask at 37 °C and 150 rpm for 24 h to obtain Lactobacillus gasseri LG08 seed liquid and Limosilactobacillus reuteri PRS-156 seed liquid.

[0046] (2) Fermentation culture: The above-activated seed liquids were respectively inoculated into 1 L of MRS medium at an inoculation amount of 1% (volume ratio) and cultured in a shaker flask at 37 °C and 150 rpm for 48 h.

[0047] The formulation of the MRS medium is as shown in Table 2 below:

[0048] Table 2

[0049] Component Content (g / L) Peptone 10.0 Beef extract 8.0 Yeast extract 4.0 Glucose 20.0 Dipotassium hydrogen phosphate 2.0 Ammonium hydrogen citrate 2.0 Sodium acetate 5.0 Magnesium sulfate 0.2 Manganese sulfate 0.04 Tween 80 1.0

[0050] Example 2

[0051] In vivo experiments were conducted on the single-strain microorganisms and microbial mixtures in Example 1 using a high-fat diet-induced obese mouse model to verify their use in treating or preventing obesity, and to compare the efficacy of the single-strain microorganisms with that of the mixed-strain microorganisms.

[0052] Experimental method:

[0053] (1) After centrifuging the cultured Lactobacillus gasseri LG08 and Lactobacillus mucosae PRS-156 bacterial solutions and discarding the supernatant, the bacterial solution concentrations were adjusted to 1×10 9 CFU / mL with sterile PBS, and the mice were gavaged according to the following groups:

[0054] Lactobacillus gasseri LG08 alone;

[0055] Lactobacillus reuteri PRS-156 alone;

[0056] A 1:1 mixture of Lactobacillus gasseri LG08 and Lactobacillus mucosae PRS-156;

[0057] The gavage volume for each group was 0.2 mL, and gavage was performed once a day.

[0058] (2) Animal experiment: 50 10-week-old C57BL / 6 female mice (purchased from Nanjing Junke Biotechnology Co., Ltd.) were purchased and fed in the same environment. 10 mice were fed a normal diet, and 40 mice were fed a high-fat diet (60% fat calories, 15% protein calories, 25% carbohydrate calories, purchased from Nantong Trofi Feed Technology Co., Ltd., feed code: TP2330055AC) for 8 weeks. The mice were weighed every 4 days. The criteria for establishing the diet-induced obesity model were that the body weight reached 28.00 ± 2.00 g. After successful modeling, the mice were grouped as follows

[0059] Group 1: Normal maintenance diet + sterile saline;

[0060] Group 2: High-fat diet + sterile saline;

[0061] Group 3: High-fat diet + Lactobacillus gasseri LG08 (viable count 1×10 9 CFU / mL, 0.2 mL gavaged per mouse);

[0062] Group 4: High-fat diet + Lactobacillus mucosae PRS-156 (viable count is 1×10 9 CFU / mL, 0.2 mL was gavaged to each mouse);

[0063] Group 5: High-fat diet + probiotic composition of Lactobacillus gasseri LG08 and Lactobacillus mucosae PRS-156 (viable count is 1×10 9 CFU / mL, the ratio of the number of strains is 1:1).

[0064] Among them, the fat calories of the normal maintenance diet is 12%, and the fat calories of the high-fat diet is 60%.

[0065] Continuous administration for 32 days, the body weight of the mice was measured every 4 days; and in the last two weeks of the intervention experiment, blood was taken from the tail vein of the mice at fixed time points every two days, the blood glucose was measured using a blood glucose meter, and the Lee's index was calculated, Lee's index = (body weight * 1000) (1 / 3) / body length (cm).

[0066] (3) Experimental results

[0067] The results are as Figure 1 - 3 shown, Figure 1 is the weight change of each group of mice every 4 days during the whole experimental process (from the 0th day of starting administration to the 32nd day of administration); Figure 2 is the relative weight change of each group of mice after administration when the modeling is successful.

[0068] From Figure 1 and Figure 2 it can be seen that there is no significant difference between Group 3 (high-fat diet + Lactobacillus gasseri LG08) and Group 2 (model group); compared with Group 2 (model group), Group 4 (high-fat diet + Lactobacillus mucosae PRS-156); compared with Group 2 (model group), Group 5 (high-fat diet + probiotic composition of Lactobacillus gasseri LG08 and Lactobacillus mucosae PRS-156) shows a greater reduction, that is, the probiotic composition can more effectively reduce the body weight of mice.

[0069] Figure 3 is the blood glucose of different groups of mice on the last day of administration, Figure 4 is the Lee's index chart of different groups of mice on the last day of administration. From Figure 3 - Figure 4 it can be seen that after starting administration, compared with Group 2, the blood glucose of Group 5 mice decreased significantly, and the Lee's index decreased significantly. There was almost no difference between Group 3 and Group 4 and no significant difference from Group 2. Combining Figure 1 and Figure 2, the body weight of the mice in Group 5 decreased most significantly, and the results showed that the single strain Lactobacillus mucosae PRS-156 in Group 4 could reduce the blood glucose and Lee's index of obese mice. However, the probiotic combination (Lactobacillus gasseri LG08 and Lactobacillus mucosae PRS-156) could more effectively reduce the body weight and blood glucose content of obese model mice, thus achieving a certain weight loss effect.

[0070] The present invention further verified the drug effects of the viable bacteria count ratios of 0.5:1 and 2:1 of Lactobacillus gasseri and Lactobacillus mucosae. The results were the same as those in Example 2. Compared with the mice in the model group, the body weight and blood glucose of these mice decreased, and the Lee's index was lower.

[0071] The results showed that the probiotic mixture provided by the implementation mode of the present invention could effectively reduce the body weight of obese mice and could be widely used in the preparation of drugs, foods or health products for regulating intestinal barrier function.

[0072] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or change without departing from the present invention shall be included within the scope of the technical solution of the present invention.

Claims

1. A probiotic composition with the effects of weight loss and blood sugar reduction, characterized in that, The raw materials of the probiotic composition include Lactobacillus gasseri and Lactobacillus mucosae reuteri.

2. The probiotic composition according to claim 1, wherein The viable count ratio of Lactobacillus gasseri to Lactobacillus mucosae reuteri is 0.5 - 2:

1.

3. The probiotic composition according to claim 1, wherein The Lactobacillus gasseri is a culture or subculture of Lactobacillus gasseri LG08.

4. The probiotic composition according to claim 1, wherein The Lactobacillus mucosae reuteri is a culture or subculture of Lactobacillus mucosae reuteri PRS - 156 with the preservation number CCTCC NO: M2023591.

5. Use of the probiotic composition according to any one of claims 1 - 4 in the preparation of a drug for treating and / or preventing obesity or obesity complications.

6. The application according to claim 5, wherein The obesity includes genetic obesity and acquired obesity; and / or, the obesity complications are selected from one or more of hypertension, hyperlipidemia, hyperglycemia, fatty liver, gallstones, cholecystitis, hyperuricemia, osteoporosis, gout, coronary heart disease, myocardial infarction, cerebral infarction, and stroke.

7. The application according to claim 6, characterized in that The acquired obesity includes: obesity caused by a high - fat diet, obesity caused by drugs, obesity caused by endocrine disorders, obesity caused by lack of sleep, obesity caused by metabolic disorders, obesity caused by mental factors, or obesity caused by aging; the obesity caused by mental factors includes: obesity caused by vagus nerve excitation or obesity caused by sympathetic nerve excitation; and / or, the obesity complication is selected from hyperglycemia.

8. A bacterial agent, characterized in that, It includes the probiotic composition according to any one of claims 1 - 4.

9. A medicament for treating and / or preventing obesity or obesity complications, characterized in that, The raw materials of the drug include the probiotic composition according to any one of claims 1 - 4 or the bacterial agent according to claim 8.

10. Use of the probiotic composition according to any one of claims 1 - 4 in the preparation of a product that helps control body fat and / or helps maintain a healthy blood glucose level.

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