A strain of Lactobacillus gasseri for relieving diabetes symptoms and its application

Abstract

The invention discloses a strain of Lactobacillus gasseri for relieving diabetes symptoms and its application, belonging to the field of microbial technology, wherein the Lactobacillus gasseri is named as Lactobacillus gasseri FLG‑219, with a deposit number of CGMCC No.28338, and has a relatively good relieving effect on various case indicator bacteria of type 2 diabetes. The probiotic composition finally provided by the invention can greatly improve the relieving effect of probiotics on two core indicators of type 2 diabetes, making the effect of probiotic preparations closer to the efficacy of drugs, which is of great significance to the clinical treatment of type 2 diabetes.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain of Lactobacillus gasseri capable of alleviating symptoms of diabetes and application thereof. Background Art

[0002] In recent years, studies have found that functional foods can improve abnormal blood sugar and blood lipid metabolism. Phytosan, which is widely distributed in vegetable oils, seeds and nuts, grains and beans, has a chemical structure similar to that of cholesterol. It can weaken cholesterol absorption through competitive inhibition and effectively reduce total cholesterol and low-density lipoprotein cholesterol in serum. Soluble dietary fiber in grains such as oats, wheat, and corn can lower blood sugar levels, improve glucose tolerance, promote bile acid metabolism, and reduce low-density lipoprotein cholesterol synthesis. Soy protein in soy milk and tofu can reduce low-density lipoprotein cholesterol and triglyceride levels in the human body.

[0003] Different from general food, functional food is a food designed and produced for specific groups of people with safety and specific health functions. It mainly regulates pathogenic genes through diet, thereby improving the symptoms and condition of related diseases, delaying and preventing the occurrence and development of complications. Functional food has the characteristics of wide sources, low toxicity and side effects, and low price, which makes it easy for people to accept it. It is the best means of daily health care and regulating physical health.

[0004] Probiotics are a type of microecological regulators of the human body. They can improve the structure of intestinal flora, promote the proliferation of beneficial bacteria in the intestine, inhibit the growth of harmful bacteria, eliminate carcinogenic factors, and improve the body's immunity. They are of great significance for the prevention and treatment of hypertension, hyperlipidemia, heart disease, diabetes and cancer. At present, there are few reports on probiotics as functional foods to improve hyperglycemia and hyperlipidemia. Therefore, the development and application of probiotics to lower blood sugar and blood lipids will become a research hotspot for functional foods and will become an important field for the development of functional foods. Probiotics refer to live microorganisms that are beneficial to the host when consumed in appropriate amounts. A large amount of evidence shows that the intake of probiotics has a beneficial effect on blood sugar through different mechanisms. Since the intestinal microbiome and its metabolites are directly involved in insulin resistance; probiotics can have a strong impact on physiological functions by changing the gastrointestinal bacterial community.

[0005] Type 2 diabetes is the most common metabolic disease in the world. Its causes are complex and cause irreversible damage to the body, which in turn induces various chronic complications. The incidence rate continues to rise worldwide. How to effectively prevent and treat diabetes has become a global health problem that needs to be solved urgently. At present, there is still no drug that can effectively cure type 2 diabetes. Patients can only control blood sugar through diet, exercise and other means, and clinical drugs also show various toxic side effects. Due to changes in lifestyle, it is difficult to achieve the expected effect by changes such as diet and exercise alone. At present, the purpose of treating diabetes is mainly achieved by taking injectable drugs. These drugs mainly include sulfonylurea and non-sulfonylurea secretagogues to stimulate pancreatic cells to secrete insulin; thiazolidinediones to enhance insulin sensitivity; biguanides to reduce liver glucose output and improve peripheral insulin resistance; and α-glucosidase inhibitors including acarbose and voglibose that can delay carbohydrate digestion and absorption. Sometimes in actual treatment, the above drugs are used in combination according to the patient's condition to achieve the purpose of rapid and effective hypoglycemia. However, the use of the above drugs will more or less produce some side effects, such as gastrointestinal discomfort, liver disease, and is limited by kidney function and heart disease. Therefore, people need to find more natural and safe blood sugar-lowering ingredients and develop safe and effective products or drugs to assist in the prevention and treatment of diabetes. In recent years, many studies have found that changes in the composition of intestinal flora play a role in the development of type 2 diabetes and obesity. In addition, it is currently recognized that the role of probiotics in diabetes management is useful.

[0006] However, in the currently disclosed technologies, we found that although probiotics can have an effect on various indicators related to diabetes symptoms, the effect is often minimal and can only play a slight auxiliary role in treatment. Even if the dosage is increased in time, it cannot achieve the desired effect. Given that the use of probiotics is one of the safe and harmless means, we need a technical solution to improve its effect, so as to upgrade probiotics from an auxiliary position in the treatment of type 2 diabetes to a therapeutic position, which is of great significance to both diabetic patients and the pharmaceutical market. Summary of the invention

[0007] In view of the above problems, the present invention provides a Lactobacillus gasseri for relieving diabetic symptoms and its application, and achieves a significant effect on diabetic symptom indicators by developing a new strain and a new composition formula used with the strain. The specific scheme is as follows:

[0008] First, the present invention provides a new Lactobacillus gasseri for relieving diabetic symptoms, named Lactobacillus gasseri FLG-219, with a preservation number of CGMCC No.28338, a preservation time of September 4, 2023, and a preservation address of the General Microbiology Center of the China Culture Collection Administration.

[0009] Second, the present invention provides the use of the above-mentioned Lactobacillus gasseri in preparing a composition for alleviating symptoms of diabetes.

[0010] The composition comprises Lactobacillus gasseri FLG-219, xylooligosaccharide and linoleic acid.

[0011] Preferably, the composition further comprises whey protein isolate.

[0012] Preferably, in the composition, the concentration of xylooligosaccharide is 3-7% (w / v).

[0013] Preferably, in the composition, the final usage concentration of linoleic acid is 0.5-1.2 g / kg.

[0014] Preferably, in the composition, the concentration of whey protein isolate is 4-9% (w / v).

[0015] Preferably, in the composition, the concentration of xylo-oligosaccharide is 5% (w / v).

[0016] Preferably, in the composition, the final usage concentration of linoleic acid is 0.5 g / kg.

[0017] Preferably, in the composition, the concentration of whey protein isolate is 5% (w / v).

[0018] In the above concentrations, (w / v) means g / mL.

[0019] In the above composition, the concentration of Lactobacillus gasseri FLG-219 is 9 x 10 7 -2x10 10 CFU.

[0020] Beneficial Effects

[0021] The beneficial effects of the present invention are:

[0022] The present invention provides a new Lactobacillus gasseri for relieving diabetes symptoms, which has a relatively good relieving effect on various case indicator bacteria of type 2 diabetes. In addition, we found that the Lactobacillus gasseri was prepared as a composition, and the synergistic effect of the composition (xylo-oligosaccharides and linoleic acid) on the metabolism of lactobacillus can greatly improve the overall hypoglycemic effect of the composition; through whey protein isolate, the effect of the probiotic composition on oxidative stress indicators is further enhanced. The probiotic composition finally provided by the present invention can greatly improve the relieving effect of probiotics on the two core indicators of type 2 diabetes, making the effect of the probiotic preparation closer to the efficacy of drugs, which is of great significance to the clinical treatment of type 2 diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is a graph showing the changes in body weight of mice in each group over time;

[0024] Figure 2 The fasting blood glucose results of each group of mice were monitored once a week for comparison;

[0025] Figure 3 It is the function of the body's pancreatic β cells and its ability to regulate blood sugar;

[0026] Figure 4-7 The insulin, insulin resistance index, β-cell function and insulin sensitivity detection index of each group of mice were respectively;

[0027] Figure 8 are the pancreatic sections of mice in each group;

[0028] Figure 9-14 They are the levels of triglyceride, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, free fatty acids in the serum of each group of mice and the calculated atherosclerosis index;

[0029] Figure 15-18 They are the contents of superoxide dismutase, reduced glutathione, glucose-6-phosphatase, and malondialdehyde in the liver of mice in each group (WPI refers to whey protein isolate);

[0030] Fig.19 is the calculated liver index of each group of mice;

[0031] Fig. 20 are the HE-stained sections of the liver of mice in each group;

[0032] Fig.21 are Oil Red O-stained sections of the liver of mice in each group;

[0033] Figure 22-23 These are the results of determination of interleukin-6 and tumor necrosis factor-α in the serum of mice in each group;

[0034] Fig.24 These are the results of HE staining of colon tissues of mice in each group. DETAILED DESCRIPTION

[0035] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0036] Unless otherwise specified, the following examples and comparative examples are parallel experiments, using the same processing steps and parameters.

[0037] Example 1 Preparation of Lactobacillus gasseri composition A:

[0038] Weigh xylooligosaccharides and dissolve them in the probiotic solution to be intragastrically administered, so that the concentration of xylooligosaccharides in the system is 5% (w / v).

[0039] Example 2 Preparation of Lactobacillus gasseri composition B:

[0040] Weigh linoleic acid and dissolve it in the probiotic solution to be intragastrically administered, so that the final intragastrically administered concentration of linoleic acid is 0.5 g / kg.

[0041] Example 3 Preparation of Lactobacillus gasseri composition C:

[0042] Weigh xylooligosaccharides and linoleic acid and dissolve them in sterile water, so that the concentration of xylooligosaccharides in the system is 5% (w / v) and the intragastric concentration of linoleic acid is 0.5 g / kg.

[0043] Example 4 Preparation of Lactobacillus gasseri composition D:

[0044] Weigh xylooligosaccharides and linoleic acid and dissolve them in the probiotic solution to be intragastrically administered, so that the concentration of xylooligosaccharides in the system is 5% (w / v) and the intragastrically administered concentration of linoleic acid is 0.5 g / kg.

[0045] Example 5 Preparation of Lactobacillus gasseri composition E:

[0046] Weigh whey protein isolate and dissolve it in the probiotic solution to be intragastrically administered, so that the concentration of whey protein isolate in the system is 5% (w / v).

[0047] In the above examples, the concentration of Lactobacillus gasseri FLG-219 is 1 x 10 9 In the following effect verification test, the FLG-219 group refers to the single feeding concentration of 1x 10 9 CFU of Lactobacillus gasseri bacterial suspension.

[0048] Effect verification:

[0049] 1. Establishment of diabetic mouse model and experimental design

[0050] Experimental animal groups:

[0051] Healthy male C57BL / 6 mice aged 6 weeks, weighing 18-20 g, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. (Beijing, China). During the experiment, the animals were given adequate food, drinking water and free movement. The animal laboratory breeding environment met the SPF grade hygiene standards (temperature 25±2℃, humidity 70%-80%), and a 12-h light-dark cycle was met during the experiment. Ordinary feed and high-sugar and high-fat feed were purchased from Jiangsu Collaborative Biotechnology Co., Ltd. (Jiangsu, China). The formula of high-sugar and high-fat feed is as follows: ordinary feed 38%, lard 28%, sucrose 5.6%, whole milk powder: 10.8%, casein 11.5%, microcrystalline cellulose: 1.9%, experimental animal premix: 2%, calcium bicarbonate 1.8%, and stone powder 0.4%.

[0052] In this experiment, a high-fat diet combined with multiple injections of low-dose STZ was used to establish a model of type 2 diabetes mice. After one week of adaptive feeding, 56 mice were randomly divided into a normal group (n=8), a model group (n=8), a FLG-219 prevention group (n=8), a composition group A, a composition group B, a composition group C, a composition group D, and a composition group E. The normal group was fed with a basic feed, and the diabetes prevention group was fed with a high-sugar and high-fat feed and given probiotics and their combination by gavage every day (1×10 9 CFU, dissolved in 300 μL of normal saline) and continued for 6 weeks. The concentration of citric acid buffer was 0, 1 mol / L and pH = 4.5. 230 mg STZ was dissolved in 100 ml of citric acid buffer. After 6 weeks, except for the normal group, the other groups were injected with STZ intraperitoneally at a dose of 45 mg / kg, once a day, for 3 consecutive days; the normal group was injected with the same volume of citric acid buffer according to body weight for control treatment. One week after STZ injection, fasting for more than 6 hours, 2 to 3 consecutive measurements of fasting blood glucose in model mice were greater than 11.1 mmol / L sugar level accompanied by polyphagia, polydipsia, polyuria and significant weight loss. Symptoms were considered to be successful diabetes modeling. Food intake, water intake, body weight and fasting blood glucose were monitored weekly during the experiment.

[0053] Probiotic intervention was continued until the day before the end of the experiment. The feces of each mouse was collected into a 2 ml sterile centrifuge tube and immediately placed in a -80 °C refrigerator for subsequent fecal metagenomic sequencing and metabolic pathway analysis; all mice were prohibited from eating feed but were free to drink sterile distilled water. After euthanasia, all mice were blooded from their orbits, left at room temperature for 1 hour to allow blood coagulation, centrifuged at 4 °C and 3500 rpm for 45 minutes, and the mouse serum was carefully collected using a pipette and stored in aliquots at -80 °C for subsequent index analysis. After dissection, the liver, kidney, intestine, and pancreas of the mice were collected and weighed, and the organ index was expressed as organ weight / body weight (mg / g). Part of the liver was placed in a 4% paraformaldehyde solution for tissue fixation, and the remaining part was quickly frozen in liquid nitrogen and stored in a -80°C refrigerator; the contents of the colon were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator, and part of the colon was placed in a 4% paraformaldehyde solution for tissue fixation, and the remaining part was quickly frozen in liquid nitrogen and stored in a -80°C refrigerator; the pancreas was placed in a 4% paraformaldehyde solution for tissue fixation.

[0054] II. Experimental study on the effects of Lactobacillus gasseri FLG-219 and its combination on glucose and lipid metabolism in diabetic mice

[0055] 1. Weight and blood sugar

[0056] During the experiment, mice were allowed to drink water and eat freely, and their food and water intake were recorded every day, and their body weight was recorded every week. After the first feeding every weekend, mice were fasted but not watered for 12 hours, and their fasting blood sugar was measured with a blood glucose meter.

[0057] 2. Oral glucose tolerance test

[0058] After the last feeding, the rats were fasted but not watered for 12 h, and then gavaged with 2 g / kg glucose solution. Blood was collected from the tail vein at 0, 15, 30, 60, and 120 min after gavage to measure blood glucose levels at that moment, and a blood glucose fluctuation curve was drawn.

[0059] The area under the glucose curve (AUC) was calculated according to the formula:

[0060] G-AUC(h*mmol / L)=(G0+G120) / 2+G15+G30+G60

[0061] 3. Sugar metabolism index

[0062] Serum insulin, glycosylated hemoglobin, glycosylated serum protein and glucagon-like peptide-1 levels were measured according to the instructions of ELISA kits, and the models for evaluating HOMA-IR and QUICKI were calculated according to the following formulas.

[0063] HOMA-IR=FBG×FINS / 22.51

[0064] QUICKI=1 / (lg FBG+lg FINS)

[0065] 4. Determination of liver gluconeogenesis and other indicators

[0066] (1) The liver was weighed and photographed and divided into two parts. The first part was sliced ​​and stained with H&E and Oil Red O for analysis and evaluation; the second part was used to measure liver cell injury indicators, including (2) in 1.5.4 and (3) in 1.5.5.

[0067] (2) The levels of HK (hexokinase), PK (pyruvate kinase), G6 (glycogen phosphorylase), and G6Pase (glucose-6-phosphatase) in the liver were determined according to the kit instructions.

[0068] 5. Lipid metabolism index

[0069] (1) Serum levels of FFA (free fatty acids), TC (total cholesterol), TG (triglycerides), LDL-C (low-density lipoprotein), and HDL-C (high-density lipoprotein) were determined according to the kit instructions.

[0070] (2) Serum levels of TNF-α, IL-6, adiponectin, leptin, and resistin were determined according to the instructions of the ELISA kit.

[0071] (3) The levels of MDA (malondialdehyde), SOD (superoxide dismutase), CAT (catalase), GSH (glutathione), and glutathione peroxidase (GSH-Px) in the liver were determined according to the kit instructions.

[0072] 3. Effect

[0073] 1. Effects of Lactobacillus gasseri FLG-219 on body weight, fasting blood glucose and OGTT in type 2 diabetic mice

[0074] Continuous weight loss is one of the typical symptoms of type 2 diabetes. Figure 1As shown in the figure, from week 0 to week 6, the Control group was fed with ordinary feed, and the Model group and FLG-219 group were fed with high-fat feed. It can be found that the weight of mice in the Control group increased the slowest, the weight of the Model group increased the most, and the weight of the FLG-219 group increased less than that of the Model group but greater than that of the normal group, which may be related to the intake of probiotics, indicating that the intake of probiotics may slow down the increase in the weight of mice caused by eating high-fat feed; after three consecutive injections of STZ in the sixth week, the weight of mice in each group decreased except for the control group; by comparing the weight values ​​of the model group at week 6 and week 7, there was a very significant difference (P < 0.0001), indicating that the diabetes modeling was successful in terms of weight. Compared with the model group, the weight loss of the FLG-219 group was less than that of the model group. The experimental results show that FLG-219 can effectively prevent the weight loss of mice with type 2 diabetes and prevent diabetic symptoms.

[0075] A high fasting blood glucose level is the most representative characteristic of type 2 diabetic mice, and the level of fasting blood glucose is also the gold standard for judging the success of the mouse diabetes model. To explore the effect of FLG-219 on blood glucose levels, fasting blood glucose was monitored once a week from the beginning of the experiment ( Figure 2 ). After STZ injection at week 6, the blood glucose level of the Control group at week 7 was significantly higher than that of the Model group (p < 0.0001), and the blood glucose levels of the FLG-219 group, oligoxylose, composition A, composition B, composition C, and composition D were significantly lower than those of the Model group (p < 0.0001), and the hypoglycemic effect of composition D was better than that of the FLG-219 group. The results showed that FLG-219 and its combination can significantly reduce the level of diabetic mice. And, from Figure 2 It can be seen that adding xylooligosaccharides alone to Lactobacillus gasseri FLG-219 will inhibit the hypoglycemic effect of the probiotics, while adding linoleic acid alone will have a slight enhancing effect on the hypoglycemic effect of the probiotics. Only when xylooligosaccharides and linoleic acid are added at the same time, the two work synergistically to greatly improve the hypoglycemic effect of the probiotic composition.

[0076] OGTT is used to evaluate the function of pancreatic β cells and their ability to regulate blood sugar ( Figure 3). After each group of mice was gavaged with glucose solution, the fasting blood glucose values ​​of each group of mice fluctuated dramatically, with a blood glucose peak at 30 minutes. From 30 minutes to 120 minutes, the blood glucose of mice in the FLG-219 group decreased fastest; at the end of the OGTT experiment, the blood glucose of mice in the FLG-219 group was significantly lower than that in the Model group (p<0.001). At the same time, compared with the Model group, the area under the OGTT curve (AUC) of the FLG-219 group was significantly reduced (p<0.05, p<0.01, p<0.05). The experimental results show that FLG-219 can significantly improve the glucose tolerance of diabetic mice.

[0077] 2. Preventive effect of Lactobacillus gasseri FLG-219 on insulin resistance in type 2 diabetic mice and pancreatic HE section analysis

[0078] Insulin is the only hormone known to the body that can lower blood sugar. Insulin resistance is a prominent feature of type 2 diabetes. To further evaluate insulin resistance, we used insulin, insulin resistance index, beta cell function, and insulin sensitivity test index to evaluate ( Figure 4-7 ). The insulin level in the Model group was significantly higher than that in the Control group (p<0.0001) and the FLG-219 group (p<0.0001). After 6 weeks of probiotic prevention, the insulin resistance index in the Model group was significantly higher than that in the Control group (p<0.0001); in terms of β-cell function, the Model group was significantly lower than the Control group (p<0.001) and the FLG-219 group (p<0.01); from the perspective of insulin sensitivity test index, the Model group was significantly lower than the Control group (p<0.0001) and the FLG-219 group (p<0.001). These results indicate that FLG-219 plays an important role in slowing down the increase in insulin in diabetic mice, slowing down the increase in insulin resistance, preventing the decline in β-cell function, and preventing the decline in the insulin sensitivity test index.

[0079] In order to further explore the changes in pancreatic islet tissue in diabetic mice, we performed HE section analysis on the pancreas of mice ( Figure 8 ). The pancreatic islet cells of the mice in the Control group were in the form of clusters, with clear edges between the islets and the surrounding tissues, dense cells in the islets, and regular arrangement. The pancreatic islet structure of the mice in the Model group was significantly damaged, the number of cells in the islets was significantly reduced and irregularly arranged, the islet tissue was atrophied, and there were vacuoles and inflammatory cell infiltration inside. After 6 weeks of probiotic intervention, the FLG-219 group effectively delayed the damage to the pancreatic islet structure, slowed down the atrophy of the islet tissue, and slowed down the phenomenon of islet cell vacuoles in diabetic mice.

[0080] 3. Effect of Lactobacillus gasseri FLG-219 on lipid metabolism in type 2 diabetic mice

[0081] Previous studies have shown that lipid metabolism is more disordered in patients with type 2 diabetes. Therefore, we measured the levels of serum triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and free fatty acids, and calculated the atherosclerosis index ( Figure 9-14 ). Compared with the Control group, the total cholesterol, triglycerides, low-density lipoprotein cholesterol and free fatty acids in the serum of the Model group were significantly increased, while the high-density lipoprotein cholesterol was significantly decreased (p < 0.0001), indicating that after 6 months of high-fat diet, the lipid metabolism ability of the Model group mice was significantly decreased. Compared with the Model group, the total cholesterol, triglycerides, low-density lipoprotein cholesterol and free fatty acids of FLG-219 were significantly reduced, while the high-density lipoprotein cholesterol was significantly increased (p < 0.01). The study showed that FLG-219 can effectively prevent lipid metabolism in diabetic mice.

[0082] 4. Effects of Lactobacillus gasseri FLG-219 on oxidative stress, HE staining and Oil Red O staining in liver tissue of type 2 diabetic mice

[0083] Long-term high-fat diet can induce fatty liver in mice, leading to liver damage and oxidative stress in the body. The mouse liver was ground and the contents of superoxide dismutase, reduced glutathione, glucose-6-phosphatase and malondialdehyde in the mouse liver were determined according to the instructions of the ELISA kit ( Figure 15-18). Superoxide dismutase is an antioxidant enzyme with free radicals as specific substrates. It removes toxins from the body by combining with free radicals and reduces the risk of free radical damage to the liver. The activity of superoxide dismutase in the liver is closely related to the health of the body. Compared with the mice in the Control group, the superoxide dismutase activity of the mice in the Model group showed a significant decrease (P < 0.0001). After 6 months of probiotic prevention, the decreasing trend of superoxide dismutase activity in the FLG-219 group was effectively controlled, and the activity was close to the normal level (p < 0.001). Glutathione can help maintain normal immune system function and has antioxidant and integrated detoxification effects. Compared with the mice in the Control group, the glutathione activity of the mice in the Model group showed a significant decrease (P < 0.0001). After 6 months of probiotic prevention, there was no significant difference between the FLG-219 group and the model group (p > 0.05). Glucose-6-phosphatase releases glucose into the blood by hydrolyzing glucose-6-phosphate in the liver tissue. Therefore, when hungry, liver glycogen can replenish blood sugar and maintain blood sugar balance. This shows that the enzyme is a key enzyme for gluconeogenesis. Compared with the control group mice, the G 6p enzyme activity of the model group mice showed a significant decrease (P < 0.0001). After 6 months of probiotic prevention, the decreasing trend of G 6p enzyme activity in the FLG-219 group was effectively controlled, and the activity was close to the normal level (P < 0.001). Compared with the control group mice, the malondialdehyde content in the liver of the Model group mice was significantly increased (P < 0.0001), and the MDA content in the FLG-219 group was significantly reduced compared with the Model group (P < 0.000). In addition, it was found that the combination E of whey protein isolate and FLG-219 can also significantly affect oxidative stress indicators. In summary, FLG-219 and its combination E can alleviate oxidative stress damage in T2DM mice.

[0084] At the same time, we calculated the liver index ( Fig.19 ), which is the weight of mouse liver (mg) / body weight (g) × 100%. The normal liver index is between 0 and 40, and a high liver index usually indicates liver damage. Compared with the Control group, the liver index of the Model group mice was significantly increased and there was a significant difference (P < 0.01); compared with the Model group, the liver index of FLG-219 was significantly reduced (P < 0.05).

[0085] The liver plays an important role in fat metabolism. To further explore the fat accumulation and liver damage in diabetic mice, we performed HE staining on the mouse liver sections ( Fig. 20 ) and Oil Red O staining analysis ( Fig.21Compared with the Control group, the liver cells of the Model group mice showed edema and necrosis, severe fat vacuoles and inflammatory cell infiltration, and disordered arrangement of liver cells; Oil Red O staining sections further showed that there was a large amount of fat accumulation in the liver of the model mice, and the area of ​​red lipid droplets increased significantly; compared with the Model group, the liver fat accumulation and red lipid droplet area of ​​the FLG-219 group were significantly reduced.

[0086] 5. Effects of Lactobacillus gasseri FLG-219 on inflammatory response and colon tissue HE staining in type 2 diabetic mice

[0087] To further explore the preventive effect of probiotics on inflammation in diabetic mice, we measured interleukin-6 and tumor necrosis factor-α in mouse serum ( Figure 22-23 ). Compared with the Control group, the levels of interleukin-6 and tumor necrosis factor-α in the serum of the Model group mice were significantly increased (****P<0.0001), while those in the FLG-219 group were significantly lower than those in the Model group (**P<0.01).

[0088] HE staining results showed that ( Fig.24 ), compared with the Control group, the Model group showed goblet cell deficiency and inflammatory cell infiltration. However, the pathological state of colon tissue in the FLG-219 group was normal, the phenomenon of goblet cell reduction was alleviated, and no inflammatory response occurred. This shows that FLG-219 can improve the pathological state of type 2 diabetic mice, reduce inflammatory response, and reduce the phenomenon of goblet cell deficiency.

[0089] The preferred specific implementation modes and embodiments of the present invention are described in detail above, but the present invention is not limited to the above implementation modes and embodiments, and various changes can be made within the knowledge scope of those skilled in the art without departing from the concept of the present invention.

Claims

1. An application of Lactobacillus gasseri for relieving diabetic symptoms in preparing a composition for relieving diabetic symptoms, characterized in that: The composition comprises Lactobacillus gasseri FLG-219, xylooligosaccharide and linoleic acid; wherein the Lactobacillus gasseri for relieving diabetes symptoms is named as Lactobacillus gasseri FLG-219, and its preservation number is CGMCC No. 28338; and the diabetes symptoms are symptoms of type II diabetes.

2. The use of Lactobacillus gasseri according to claim 1 in preparing a composition for alleviating diabetes symptoms, characterized in that: Whey protein isolate is also included in the composition.

3. The use of Lactobacillus gasseri according to claim 1 in preparing a composition for alleviating diabetes symptoms, characterized in that: In the composition, the concentration of xylo-oligosaccharide is 3-7% (w / v).

4. The use of Lactobacillus gasseri in preparing a composition for alleviating diabetes symptoms according to claim 1, characterized in that: In the composition, the final concentration of linoleic acid is 0.5-1.2 g / kg.

5. The use of Lactobacillus gasseri in preparing a composition for alleviating diabetes symptoms according to claim 2, characterized in that: In the composition, the concentration of whey protein isolate is 4-9% (w / v).

6. The use of Lactobacillus gasseri in preparing a composition for alleviating diabetes symptoms according to claim 1, characterized in that: In the composition, the concentration of xylo-oligosaccharide is 5% (w / v).

7. The use of Lactobacillus gasseri in preparing a composition for alleviating diabetes symptoms according to claim 1, characterized in that: In the composition, the final concentration of linoleic acid is 0.5 g / kg.

8. The use of Lactobacillus gasseri in preparing a composition for alleviating diabetes symptoms according to claim 2, characterized in that: In the composition, the concentration of whey protein isolate is 5% (w / v).

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