Application of bifidobacterium animalis subsp. Lactis BLa80 in preparation of preparation for assisting in improving diabetes mellitus

Abstract

The invention relates to application of bifidobacterium animalis subsp. Lactis BLa80 in preparation of a preparation for assisting in improving diabetes mellitus. The bifidobacterium animalis subsp. Lactis BLa80 is a strain of the bifidobacterium animalis subsp. Lactis BLa80 with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.22547, and the strain of the bifidobacterium animalis subsp. Lactis BLa80 is a strain of the bifidobacterium animalis subsp. Lactis BLa80 with the preservation number of CGMCC No.22547. The brand new application of the BLa80 strain of the animal bifidobacterium subsp. Lactis is developed, that is, the BLa80 strain is used for improving diabetes, reducing blood sugar or inhibiting alpha-glucosidase, and tests prove that the BLa80 strain can effectively control the food intake, the water intake, the urine volume and the body weight of diabetic mice; the fasting blood-glucose value and the insulin resistance index of a diabetic mouse are reduced; the blood lipid level disorder state of diabetic mice can be improved; the inflammation level of a diabetic mouse body is reduced, anti-inflammatory factors of the mouse body are increased, and the immunity is improved; and the abundance and diversity of intestinal flora of diabetic mice can be improved and recovered.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and relates to a new use of Bifidobacterium animalis subsp. lactis BLa80, in particular to the application of Bifidobacterium animalis subsp. lactis BLa80 in the preparation of a preparation for assisting in improving diabetes. Background Art

[0002] Diabetes is a systemic metabolic disease characterized by chronic elevation of blood glucose levels, which is caused by insufficient insulin secretion or insulin resistance resulting from the combined action of multiple environmental factors and genetic factors. If diabetes is not actively treated, a series of complications will occur over time, such as cardiovascular and cerebrovascular diseases, retinopathy, diabetic nephropathy, neuropathy, acral necrosis, etc. With the development of the economy and the change of diet structure, the incidence of diabetes has been increasing year by year. At present, diabetes is still a disease that cannot be completely cured. Four-level preventive measures are taken for the prevention of diabetes, and primary prevention is the most important, aiming to reduce the incidence of diabetes. Clinically, although it is recognized that a combination of diet therapy, exercise therapy and drug therapy is used to treat diabetes, generally drug therapy is still the main method, mainly insulin and oral hypoglycemic drugs, which are roughly divided into sulfonylureas, biguanides, traditional Chinese medicine preparations, other hypoglycemic drugs and adjuvant drugs, to control blood glucose elevation, relieve the symptoms of diabetes, and delay the further deterioration of diabetes and the occurrence of complications.

[0003] Studies have shown that there are differences in the composition of the intestinal flora among healthy people, obese people and type 2 diabetes patients, suggesting that the intestinal flora may be a factor affecting the pathophysiology of metabolic diseases in the body. Therefore, it is an effective method to prevent or treat metabolic diseases caused by high-fat diet by effectively regulating the intestinal flora. Probiotics are active microorganisms ingested by the body, which can effectively improve the balance of the intestinal flora and produce good health effects on the host. Bifidobacterium animalis subsp. lactis is a type of Gram-positive bacillus, mainly present in the small intestine, releasing lactic acid, acetic acid and some antibiotics that act on harmful bacteria. Bifidobacterium animalis subsp. lactis in the human intestine plays an important role in human health. However, there are relatively few Bifidobacterium animalis subsp. lactis that can safely and effectively lower blood glucose and improve diabetes. Therefore, more Bifidobacterium animalis subsp. lactis with α-glucosidase inhibitory ability and blood glucose lowering effect need to be developed in this field. Summary of the Invention

[0004] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a new use of Bifidobacterium animalis subsp. lactis BLa80, in particular to provide the application of Bifidobacterium animalis subsp. lactis BLa80 in the preparation of a preparation for assisting in improving diabetes.

[0005] To achieve the object of the present invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides the use of Bifidobacterium animalis subsp. lactis BLa80 in the preparation of a preparation for assisting in improving diabetes;

[0007] The Bifidobacterium animalis subsp. lactis BLa80 is the Bifidobacterium animalis subsp. lactis BLa80 strain with the preservation number of CGMCC No. 22547.

[0008] The present invention has developed a new use of the Bifidobacterium animalis subsp. lactis BLa80 strain, that is, using it to improve diabetes, lower blood sugar or inhibit α-glucosidase. Experiments have proved that the BLa80 strain can effectively control the food intake, water intake, urine volume and body weight of diabetic mice; and can reduce the fasting blood glucose value and insulin resistance index of diabetic mice, indicating that the BLa80 strain can improve the glucose absorption and metabolism ability of diabetic mice, slow down the damage of pancreatic islet β-cell function, and thus effectively reduce the blood glucose value of mice; at the same time, it can also improve the disordered state of blood lipid levels in diabetic mice; reduce the inflammation level of the diabetic mouse body, increase the anti-inflammatory factors in the mouse body, and improve immunity; it can also improve and restore the abundance and diversity of the intestinal flora of diabetic mice.

[0009] Preferably, the viable bacteria count in the preparation is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g, such as 1×10 8 CFU / mL (CFU / g), 5×10 8 CFU / mL (CFU / g), 1×10 9 CFU / mL (CFU / g), 5×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), etc.

[0010] Preferably, the dosage form of the preparation includes solution, powder, tablet or capsule.

[0011] Preferably, the preparation further contains excipients; the excipients include any one or a combination of at least two of carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, bacteriostatic agents or buffers.

[0012] Preferably, the active components in the preparation include Bifidobacterium animalis subsp. lactis BLa80 and metformin or its salt.

[0013] The above-mentioned BLa80 strain can not only be used alone to assist in improving diabetes or reducing blood sugar, but can also be used in combination with the drug metformin or its salt. Metformin is a first-line drug for the treatment of type II diabetes.

[0014] In the present invention, the Bifidobacterium animalis subsp. lactis BLa80 strain and the drug metformin hydrochloride are combined and used in combination, and it is found that the two can cooperate with each other and promote each other, and have a synergistic effect in terms of reducing blood sugar and improving diabetes. Compared with the single BLa80 strain intervention method or the single metformin hydrochloride intervention method, the combined use of the two active components can improve the effects of reducing blood sugar and improving diabetes with a lower intervention amount.

[0015] Preferably, the usage ratio of the Bifidobacterium animalis subsp. lactis BLa80 to metformin or its salt is (10 7 -10 9 ) CFU: (1-5) μg.

[0016] Among them, the specific point values in (10 7 -10 9 ) can be selected as 1×10 7 , 3×10 7 , 5×10 7 , 8×10 7 , 1×10 8 , 2×10 8 , 4×10 8 , 6×10 8 , 8×10 8 , 1×10 9 and so on.

[0017] Among them, the specific point values in (1-5) can be selected as 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 and so on.

[0018] Based on the potential synergistic cooperation relationship between the two active components, when it meets the above specific usage ratio, its effect in reducing blood sugar and improving diabetes is more excellent.

[0019] In the second aspect, the present invention provides the use of Bifidobacterium animalis subsp. lactis BLa80 in the preparation of an α-glucosidase inhibitor;

[0020] The Bifidobacterium animalis subsp. lactis BLa80 is the Bifidobacterium animalis subsp. lactis (Bifidobacterium animalis subsp. lactis) BLa80 strain with the preservation number of CGMCC No. 22547.

[0021] The present invention also developed another use of Bifidobacterium animalis subsp. lactis strain BLa80, that is, using it as an α-glucosidase inhibitor. According to the research results of the present invention, Bifidobacterium animalis subsp. lactis strain BLa80 has an excellent effect of inhibiting α-glucosidase. Therefore, this result indicates that Bifidobacterium animalis subsp. lactis strain BLa80 can also be used as a simple reagent for in vitro experiments in the scientific research field.

[0022] In a third aspect, the present invention provides the use of Bifidobacterium animalis subsp. lactis BLa80 in the preparation of a product having any one or at least two of the following effects:

[0023] 1) Improving the ability of glucose absorption and metabolism; 2) Slowing down the impairment of pancreatic islet β-cell function in diabetes; 3) Improving the disorder of blood lipids in diabetes; 4) Reducing the inflammatory level and enhancing immunity; 5) Improving the richness and diversity of the intestinal flora in the diabetic body;

[0024] The Bifidobacterium animalis subsp. lactis BLa80 is the Bifidobacterium animalis subsp. lactis BLa80 strain with the preservation number of CGMCC No. 22547.

[0025] In a fourth aspect, the present invention provides a probiotic composition having a hypoglycemic effect, and the active components of the probiotic composition include Bifidobacterium animalis subsp. lactis BLa80 and metformin or its salt.

[0026] The present invention creatively combines Bifidobacterium animalis subsp. lactis strain BLa80 and metformin or its salt to obtain a composition product. It is found that this composition can not only reduce the intervention doses of Bifidobacterium animalis subsp. lactis BLa80 and metformin or its salt, improve the drug safety, but also has a more significant effect on improving diabetes than single Bifidobacterium animalis subsp. lactis BLa80 and metformin or its salt, showing a synergistic promotion effect. The present invention provides an effective combination strategy for the improvement of diabetes, which has very significant significance.

[0027] Preferably, the usage ratio of the Bifidobacterium animalis subsp. lactis BLa80 to metformin or its salt is (10^7 - 10^9) CFU : (1 - 5) μg.

[0028] Among them, the specific point values in (10 7 - 10 9 ) can be selected as 1×10 7 , 3×10 7 , 5×10 7 , 8×10 7 、1×108 , 2×10 8 , 4×10 8 , 6×10 8 , 8×10 8 , 1×10 9 etc.

[0029] Among them, the specific point values in (1-5) can be selected as 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, etc.

[0030] In the fifth aspect, the present invention provides the application of the probiotic composition described in the fourth aspect in the preparation of a product for preventing and treating diabetes or assisting in improving diabetes.

[0031] All other point values not specifically listed in all the numerical ranges involved in the present invention are within the protection scope of the present invention. Considering space and brevity of description, they will not be elaborated one by one here.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The present invention has developed a new use of the Bifidobacterium animalis subsp. lactis BLa80 strain, that is, using it to improve diabetes, lower blood sugar or inhibit α-glucosidase. Experiments have proved that the BLa80 strain can effectively control the food intake, water intake, urine output and body weight of diabetic mice; and can reduce the fasting blood sugar value and insulin resistance index of diabetic mice, indicating that the BLa80 strain can improve the glucose absorption and metabolism ability of diabetic mice, slow down the damage of pancreatic islet β-cell function, thereby effectively reducing the blood sugar value of mice; at the same time, it can also improve the disordered state of blood lipid levels in diabetic mice; reduce the inflammation level of the diabetic mouse body, increase the anti-inflammatory factors in the mouse body, and improve immunity; it can also improve and restore the abundance and diversity of the intestinal flora of diabetic mice.

[0034] Furthermore, the present invention combines and uses the Bifidobacterium animalis subsp. lactis BLa80 strain and the drug metformin hydrochloride, and finds that the two can cooperate with each other and promote each other, and have a synergistic effect in terms of blood sugar lowering and diabetes improvement. Compared with the single BLa80 strain intervention method or the single metformin hydrochloride intervention method, the combination of the two active components can improve the blood sugar lowering and diabetes improvement effects with a lower intervention amount. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 is a statistical result graph of the average food intake of each group of mice;

[0036] Figure 2 is a statistical result graph of the average water intake of each group of mice;

[0037] Figure 3It is a statistical result graph of the average body weight of each group of mice;

[0038] Figure 4 It is a measurement result graph of the fasting blood glucose value (FBG) of each group of mice;

[0039] Figure 5 It is a statistical result graph of the area under the oral glucose tolerance curve of each group of mice;

[0040] Figure 6 It is a statistical result graph of the insulin resistance index of each group of mice;

[0041] Figure 7 It is a statistical result graph of the serum LPS level of each group of mice;

[0042] Figure 8 It is a statistical result graph of the serum TNF-α level of each group of mice;

[0043] Figure 9 It is a statistical result graph of the serum IL-6 level of each group of mice;

[0044] Figure 10 It is a statistical result graph of the serum IL-10 level of each group of mice;

[0045] Figure 11 It is a statistical result graph of the Chao1 index and Simpson index of the intestinal flora of each group of mice;

[0046] Figure 12 It is a NMDS analysis result graph of the intestinal flora of each group of mice;

[0047] Figure 13 It is a statistical result graph of the relative abundance of the intestinal flora of each group of mice;

[0048] The taxonomic nomenclature of the BLa80 strain involved in the present invention is Bifidobacterium animalis subsp. lactis, the preservation time is May 17, 2021, the preservation number is CGMCC No. 22547, the preservation unit is the China General Microbiological Culture Collection Center, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Detailed implementation manners

[0049] The technical solutions of the present invention will be further described below through specific implementation manners. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations on the present invention.

[0050] The taxonomic naming of the BLa80 strain involved below is Bifidobacterium animalis subsp. lactis, the preservation time is May 17, 2021, and the preservation number is CGMCC No. 22547.

[0051] The preparation method of the BLa80 bacterial suspension involved in the following experiment: Inoculate the BLa80 strain into L-MRS liquid medium, culture it at 37 °C for 18 h for activation, and activate it continuously twice to obtain the activation solution; inoculate the activation solution into L-MRS liquid medium at an inoculation amount of 3% (v / v), culture it at 37 °C for 24 h to obtain the bacterial solution; centrifuge the bacterial solution at 4000 rpm at 4 °C for 5 min, filter it to obtain the bacterial cells, resuspend the bacterial cells with physiological saline and dilute them as needed to obtain the BLa80 bacterial suspension.

[0052] L-MRS medium: Peptone 10 g / L, Beef extract 10 g / L, Glucose 20 g / L, Sodium acetate 2 g / L, Yeast extract 5 g / L, Diammonium hydrogen citrate 2 g / L, K 2 PO 4 ·3H 2 O 2.6 g / L, MgSO 4 ·7H 2 O 0.1 g / L, MnSO 4 0.05 g / L, Tween 80 1 mL / L, Cysteine hydrochloride 0.5 g / L.

[0053] Example 1

[0054] Evaluation of the inhibitory effect of Bifidobacterium animalis subsp. lactis BLa80 on α-glucosidase:

[0055] Adjust the concentration of the bacterial suspension to 10 8 CFU / mL for standby. Add 150 μL of PBS solution to 75 μL of p-nitrophenyl-α-D-glucopyranoside (PNPG) solution and 25 μL of the test sample bacterial suspension or PBS solution, place the mixture on ice for 3 min, then add 50 μL of α-glucosidase solution (0.2 U / mL), react in a water bath at 37 °C for 15 min, add 1 mL of a 1 mol / L Na 2 CO 3 As the reaction termination solution, measure the absorbance value of the reaction solution at 405 nm, and then calculate the α-glucosidase inhibition rate (%) by substituting the absorbance value into the following formula.

[0056] α-glucosidase inhibition rate = [1 - (A - B) / (C - D)] × 100%

[0057] Among them, A is the absorbance value of the sample added with the bacterial suspension and α-glucosidase solution; B is the absorbance value of the sample added with the bacterial suspension but without α-glucosidase solution; C is the absorbance value of the sample without the bacterial suspension but added with α-glucosidase solution; D is the absorbance value of the sample without the bacterial suspension and without α-glucosidase solution.

[0058] According to the above formula, the α-glucosidase inhibition rate of Bifidobacterium animalis subsp. lactis BLa80 was calculated to be (16.91 ± 0.07)%. It can be seen from this that Bifidobacterium animalis subsp. lactis BLa80 has the ability to inhibit α-glucosidase, indicating that this strain has potential hypoglycemic ability.

[0059] Example 2

[0060] Evaluation of the improvement effect of Bifidobacterium animalis subsp. lactis BLa80 on type II diabetic mice:

[0061] (1) Experimental animals: Healthy male C57BL / 6j mice (5 - 6 weeks old; 19 ± 1 g) were housed in the animal house of Hubei Center for Disease Control and Prevention. The experimental animal use license number was: SCXK(E)2020 - 0019. These mice were housed in a controlled environment with the room temperature maintained at 20 - 22 °C, the humidity at 40 - 60%, following a 12 h light / dark cycle. They had free access to food and water. The bedding was changed 1 - 2 times a week.

[0062] (2) Animal grouping: After 1 week of adaptive feeding, the mice were randomly assigned to 9 groups, with 8 mice in each group: normal group (NC group), model group (MC group), probiotic BLa80 group (BLa80 group, intervened with BLa80 bacterial solution), metformin hydrochloride group (MET group, intervened with metformin hydrochloride), probiotic ATCC700541 group (ATCC700541 group, intervened with commercially available Bifidobacterium animalis subsp. lactis bacterial solution), BLa80 + MET groups 1 - 3 (co - intervened with different ratios of BLa80 bacterial solution and metformin hydrochloride), ATCC700541 + MET group (co - intervened with commercially available Bifidobacterium animalis subsp. lactis bacterial solution and metformin hydrochloride).

[0063] (3) Animal modeling and intervention methods:

[0064] The type 2 diabetes model was established in mice according to the method described in the reference (F. Yan, N. Li, J. Shi, H. Li, Y. Yue, W. Jiao, N. Wang, Y. Song, G. Huo and B. Li, Lactobacillus acidophilus alleviates type 2 diabetes by regulating 2 hepatic glucose and lipid metabolism and gut microbiota in mice, Food Funct, 2019,).

[0065] After successful modeling, the mice in each group were intervened for 35 days. Among them, the mice in the normal group (NC group) and the model group (MC group) were gavaged with sterile normal saline once a day for 35 consecutive days;

[0066] The BLa80 group was gavaged with the suspension of Bifidobacterium animalis subsp. lactis BLa80 (the intervention dose was 6×10 9 CFU / d) once a day for 35 consecutive days;

[0067] The ATCC700541 group was gavaged with the suspension of Bifidobacterium animalis subsp. lactis ATCC700541 (the intervention dose was 6×10 9 CFU / d) once a day for 35 consecutive days;

[0068] The MET group was gavaged with metformin hydrochloride (MET) (the intervention dose was 0.2 g / kg / d) once a day for 35 consecutive days;

[0069] The BLa80+MET group 1 was gavaged with the suspension of Bifidobacterium animalis subsp. lactis BLa80 (the intervention dose was 6×10 7 CFU / d) and metformin hydrochloride (MET) (the intervention dose was 0.15 g / kg / d) once a day for 35 consecutive days;

[0070] The BLa80+MET group 2 was gavaged with the suspension of Bifidobacterium animalis subsp. lactis BLa80 (the intervention dose was 6×10 8 CFU / d) and metformin hydrochloride (MET) (the intervention dose was 0.15 g / kg / d) once a day for 35 consecutive days;

[0071] The BLa80+MET group 3 was gavaged with the suspension of Bifidobacterium animalis subsp. lactis BLa80 (the intervention dose was 6×10 9 CFU / d) and metformin hydrochloride (MET) (the intervention dose was 0.15 g / kg / d) once a day for 35 consecutive days;

[0072] The ATCC700541 + MET group was intragastrically administered with an ATCC700541 bacterial suspension (intervention dose: 6×10 9 CFU / d) and metformin hydrochloride (MET) (intervention dose: 0.15 g / kg / d) once a day for 35 consecutive days.

[0073] (4) Index analysis:

[0074] (4.1) Monitoring of food intake, water intake, and body weight:

[0075] During the intervention period, the average food intake, average water intake, and average body weight of mice in each group were measured and statistically analyzed every week. The statistical results are as Figure 1 、 Figure 2 and Figure 3 shown.

[0076] It can be seen that compared with the normal group, the mice in the type 2 diabetes model group showed an increase in daily food intake, an increase in water intake, an increase in urine output, and a decrease in body weight. After five weeks of intervention treatment in each group, the typical symptoms of type 2 diabetic mice were alleviated and improved to varying degrees. Specifically, the food intake and water intake of the mice were controlled, the body weight of the mice recovered, and the effects of groups 1 - 3 in the BLa80 + MET group were better, indicating that when the BLa80 strain and metformin hydrochloride are used in combination, not only can the intervention doses of each be reduced, but also they have a more significant effect on improving diabetes than a single BLa80 bacterial solution or metformin hydrochloride, playing a synergistic promoting role.

[0077] (2) Effects on glucose metabolism:

[0078] (2.1) Fasting blood glucose value: Before treatment and during the five - week treatment period, blood was collected from the tail veins of mice every week to measure the fasting blood glucose value of the mice. The measurement results of the fasting blood glucose value (FBG) of mice in each group are as Figure 4 shown.

[0079] It can be seen that after five weeks of intervention, the fasting blood glucose values of each group of type 2 diabetic model mice decreased to varying degrees and approached the healthy group. Among them, the effects of groups 1 - 3 in the BLa80 + MET group were better, indicating that when the BLa80 strain and metformin hydrochloride are used in combination, not only can the intervention doses of each be reduced, but also they have a more significant effect on improving diabetes than a single BLa80 bacterial solution or metformin hydrochloride, playing a synergistic promoting role.

[0080] (2.2) Oral glucose tolerance: After five weeks of treatment, oral glucose tolerance test was performed on mice. The glucose dosage for the glucose tolerance test in mice was 2 g per kilogram of body weight (2 g / kg), and a 20% glucose solution was prepared with normal saline. Mice were fasted for 16 hours while maintaining normal drinking water. Before starting the glucose tolerance test, the weight of each mouse was measured. Blood was taken from the tip of the mouse's tail, and the fasting blood glucose was measured with a blood glucose meter. The measured value was regarded as the blood glucose value at 0 min. Glucose was administered by gavage and timing started immediately after completion. The blood glucose values of each mouse were measured at 15, 30, 60, 90, and 120 min. The area under the oral glucose tolerance curve was calculated, and the statistical results are as Figure 5 shown.

[0081] It can be seen that the area under the oral glucose tolerance curve of each intervention group increased to varying degrees compared with the model group and approached the healthy value, indicating that the BLa80 strain can effectively improve the glucose absorption and metabolism ability of diabetic mice, thereby showing a blood glucose lowering effect. The combination of BLa80 and MET was more prominent in the above effects.

[0082] (2.3) Insulin resistance index: The insulin resistance index (HOMA-IR) is a value calculated according to the serum insulin level and fasting blood glucose value, which is an index used to judge the degree of insulin resistance and can effectively reflect the functional state of pancreatic islet β cells in the body. The statistical results are as Figure 6 shown.

[0083] It can be seen that the insulin resistance indexes of mice in the BLa80 group, MET group, and BLa80+MET groups 1-3 were significantly lower than those in the model group and approached the healthy value, indicating that the intervention of the BLa80 strain can effectively slow down the continuous damage of pancreatic islet β cell function, thereby effectively reducing the blood glucose value of mice. The combination of the BLa80 strain and MET was more prominent in the above effects.

[0084] (3) Effects on blood lipid levels:

[0085] After the intervention, the following serum indexes of each group of mice were measured, including: triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). The results are shown in Table 1 (where different letters after the data represent significant differences between groups, p < 0.05).

[0086] Table 1

[0087]

[0088]

[0089] As can be seen from the data results in Table 1, compared with the mice in the model group, the blood lipid levels of the mice in each intervention group were regulated and improved to varying degrees. Moreover, the effects of groups BLa80, MET, and BLa80+MET in 1-3 were more excellent, and the group BLa80+MET in 1-3 was the best. This indicates that in the treatment of diabetic dyslipidemia, strain BLa80 has an obvious effect on reducing total cholesterol and high-density lipoprotein in the serum. The effect is even more significant after combination with MET, suggesting that strain BLa80 and MET have a synergistic effect.

[0090] (4) Effects on inflammatory factors:

[0091] After the intervention ended, whole blood of the mice in each group was centrifuged to obtain serum, and the inflammatory factors in the mouse serum were measured using an ELISA kit, including the concentration levels of LPS (lipopolysaccharide), TNF-α, IL-6, and IL-10, as shown respectively in Figure 7 、 Figure 8 、 Figure 9 、 Figure 10 . As can be seen from the figure, compared with the model group (MC), the levels of inflammatory factors in the mice in each intervention group were controlled to varying degrees, and the levels of anti-inflammatory factors were effectively increased. This indicates that strain BLa80 can relieve the in vivo inflammation of type 2 diabetic mice and improve the immunity of type 2 diabetic mice. The effect after combination with MET is better than that of using strain BLa80 alone or MET alone, that is, strain BLa80 and MET have a synergistic effect.

[0092] (5) Effects on gut microbiota:

[0093] After the intervention ended, the mice were sacrificed. After abdominal disinfection, the mice were dissected, and the cecal contents of the mice in the normal group (NC group), model group (MC group), MET group, and BLa80 group were taken for microbial diversity analysis. The V3-V4 hypervariable region of the 16S rDNA of all bacteria in the samples was sequenced on the MiSeq Illumina sequencing platform to determine the characteristics of the gut microbiota.

[0094] α-diversity reflects the species richness and species diversity of the sample. The Chao1 index and Simpson index are commonly used judgment indicators for the abundance and diversity of gut microbial colonies. The results are shown in Figure 11As shown, the results indicate that the occurrence of type Ⅱ diabetes can lead to a decrease in the richness and diversity of the gut microbiota in the body. The intragastric administration of strain BLa80 can significantly increase the abundance of the gut microbiota, which is comparable to the level of drug treatment and shows no significant difference (p>0.05). According to the analysis of Simpson index data, intragastric administration of strain BLa80 can extremely significantly increase the diversity of the gut microbiota in type Ⅱ diabetic mice and approach that of the healthy group. In summary, five weeks of intragastric administration of strain BLa80 can play a good role in improving and restoring the decrease in the abundance and diversity of the gut microbiota in type Ⅱ diabetic mice.

[0095] β-diversity is used to compare the similarity in species diversity among different samples. The NMDS method is a common analysis method that can reflect the differences and distances between samples. When Stress is less than 0.2, it indicates that the NMDS analysis has a certain degree of reliability. The closer the samples are on the coordinate graph, the higher their similarity. The results of each group are as Figure 12 shown. It can be seen that the treatment with probiotics and drugs can improve the gut microbiota of type Ⅱ diabetic mice and show differences from the model group, while approaching the healthy mice. This shows that intragastric administration of strain BLa80 for five weeks can effectively improve the gut microbiota of type Ⅱ diabetic mice and tend to be normal.

[0096] The relative abundances of different genera of bacteria in the intestines of each group of mice are as Figure 13 shown. It was found that the genera Blautia and Bacteroides were significantly increased in the gut microbiota of mice administered BLa80 intragastrically. Literature reports that the genus Bacteroides is involved in the metabolism of glucose and other carbohydrates and produces short-chain fatty acids (such as propionic acid and acetic acid), which are beneficial to gut health; the abundance of Allobaculum in the bodies of mice administered BLa80 intragastrically for five weeks was extremely significantly increased, and it can utilize carbohydrates to generate butyric acid, indicating that BLa80 can effectively regulate the health of the gut microbiota; in addition, the beneficial gut bacteria Akkermansia muciniphila and unclassified_Muribaculaceae showed significant increases in abundance in the intervention group of strain BLa80 (p<0.05); the harmful gut bacteria Aerococcus showed extremely significant decreases in the BLa80 treatment group and the drug treatment group (p<0.01).

[0097] The applicant declares that the technical solution of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0098] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0099] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any appropriate manner. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. Application of Bifidobacterium animalis subspecies lactis BLa80 in the preparation of a preparation for assisting in improving diabetes; The Bifidobacterium animalis subsp. lactis BLa80 is a strain of Bifidobacterium animalis subsp. lactis BLa80 with a preservation number of CGMCC No.22547.

2. The use according to claim 1, characterized in that: The number of viable bacteria in the preparation is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g.

3. The use according to claim 1, characterized in that: The dosage form of the preparation includes solution, powder, tablet or capsule; Preferably, the preparation further contains excipients; the excipients include any one or a combination of at least two of carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

4. The use according to claim 1, characterized in that: The active ingredients in the preparation include Bifidobacterium animalis subspecies lactis BLa80 and metformin or a salt thereof.

5. The use according to claim 4, characterized in that: The ratio of animal Bifidobacterium lactis subspecies BLa80 to metformin or its salt is (10 7 -10 9 )CFU:(1-5)μg.

6. Application of Bifidobacterium animalis subspecies lactis BLa80 in the preparation of α-glucosidase inhibitors; The Bifidobacterium animalis subsp. lactis BLa80 is a strain of Bifidobacterium animalis subsp. lactis BLa80 with a preservation number of CGMCC No.22547.

7. Use of Bifidobacterium animalis subspecies lactis BLa80 in the preparation of a product having any one or at least two of the following effects: 1) Improve glucose absorption and metabolism; 2) Slow down the damage of diabetic pancreatic β-cell function; 3) Improve diabetic dyslipidemia; 4) Reduce inflammation level and improve immunity; 5) Improve the richness and diversity of intestinal flora in diabetic patients; The Bifidobacterium animalis subsp. lactis BLa80 is a strain of Bifidobacterium animalis subsp. lactis BLa80 with a preservation number of CGMCC No.22547.

8. A probiotic composition having a blood sugar lowering effect, characterized in that: The active components of the probiotic composition include Bifidobacterium animalis subspecies lactis BLa80 and metformin or a salt thereof.

9. The probiotic composition according to claim 7, characterized in that The ratio of animal Bifidobacterium lactis subspecies BLa80 to metformin or its salt is (10 7 -10 9 )CFU:(1-5)μg.

10. Use of the probiotic composition according to claim 8 or 9 in preparing a product for preventing and treating diabetes or assisting in improving diabetes.

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