Phytobacterium plantarum Lp116 with hypoglycemic capacity and application, product and method thereof

Abstract

The invention relates to a plant lactobacillus with hypoglycemic ability, and an application, a product and a method thereof, and belongs to the technical field of microorganisms. The invention provides a lactobacillus plantarum strain Lp116 with the capability of reducing blood sugar, and the preservation number of the lactobacillus plantarum strain Lp116 is CCTCC (China Center for Type Culture Collection) NO: M 20241784. The invention also provides an application of the strain Lp116 in preparation of hypoglycemic drugs or health-care foods with an auxiliary hypoglycemic function or bacteriostatic agents or non-treatment-destination bacteriostasis, and a product and a method based on the strain Lp116. The strain Lp116 disclosed by the invention has a relatively strong inhibition effect on escherichia coli, staphylococcus aureus, salmonella, streptococcus pharyngobuccal inflammation and streptococcus pseudopneumoniae, and can effectively achieve the effects of reducing blood sugar and regulating intestinal flora.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to a Lactobacillus plantarum Lp116 capable of lowering blood sugar, and an application, product and method thereof. Background Art

[0002] Diabetes has become a serious global public health problem, and the prevalence of diabetes in China is particularly severe. Driven by factors such as rapid economic development and accelerated industrialization, changes in lifestyle and aging, the prevalence of diabetes in my country has been increasing year by year, bringing heavy social and economic burdens. Type 2 diabetes is a chronic metabolic disease caused by insufficient or abnormal insulin secretion in insulin-sensitive tissues, which leads to increased blood sugar. Type 2 diabetes accounts for 95% of all diabetic patients.

[0003] Lactobacillus plantarum is a type of Gram-positive lactobacillus that is widely found in nature, especially in various fermented foods such as vegetables, meat, dairy products and wine. At the same time, Lactobacillus plantarum also exists in the human gastrointestinal tract and is a probiotic bacteria in the human gastrointestinal tract, which has a great promoting effect on human health.

[0004] At present, Lactobacillus plantarum with strong α-glucosidase inhibition ability and both hypoglycemic effect and broad-spectrum antibacterial properties is very rare in the art. Therefore, the art is in urgent need of developing new Lactobacillus plantarum strains. Summary of the invention

[0005] In order to solve the technical problem that there is a lack of Lactobacillus plantarum with strong α-glucosidase inhibition ability and both blood sugar lowering effect and broad-spectrum antibacterial properties in the art, the present invention aims to provide a Lactobacillus plantarum Lp116 with blood sugar lowering ability and its application, product and method.

[0006] The technical solution of the present invention is as follows:

[0007] A Lactobacillus plantarum Lp116 having blood sugar lowering ability, characterized in that its preservation number is CCTCC NO:M 20241784.

[0008] The Lactobacillus plantarum Lp116 with the deposit number of CCTCC NO: M 20241784 is used for preparing hypoglycemic drugs or health foods with auxiliary hypoglycemic function or antibacterial agents or antibacterial agents for non-therapeutic purposes.

[0009] The antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis and Streptococcus pseudopneumoniae.

[0010] The hypoglycemic drug uses Lactobacillus plantarum Lp116 as a pharmaceutically active ingredient.

[0011] A hypoglycemic drug, comprising a pharmacologically active ingredient, characterized in that the pharmacologically active ingredient comprises: Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784.

[0012] The hypoglycemic drug further comprises: pharmaceutically acceptable excipients.

[0013] A health food with auxiliary blood sugar lowering function, comprising: Lactobacillus plantarum Lp116 with the preservation number of CCTCC NO: M20241784.

[0014] The health food with auxiliary blood sugar lowering function also includes auxiliary materials.

[0015] A non-therapeutic antibacterial method adopts Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 for antibacterial purpose.

[0016] The antibacterial agent refers to: inhibiting Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis, and Streptococcus pseudopneumoniae.

[0017] The beneficial effects of the present invention are as follows:

[0018] The invention provides a plant lactobacillus (Lactiplantibacillus plantarum) Lp116 with blood sugar lowering ability. The tolerance survival rate of the strain to artificial gastric juice and bile salt is 96.85% and 92.53% respectively, the inhibition rate to alpha-glucosidase is 16.00%, the diet, water intake and body weight of type II diabetic mice have a tendency to return to normal, the sugar metabolism ability of the mice can be effectively improved and enhanced, the blood sugar value of the mice is reduced, and the blood lipid level of the type II diabetic mice can be effectively reduced. The plant lactobacillus (Lactiplantibacillus plantarum) Lp116 of the invention can also simultaneously inhibit pathogens such as Escherichia coli, Salmonella, Staphylococcus aureus, Streptococcus pharyngitis, Streptococcus pseudopneumoniae, etc., and has a broad-spectrum antibacterial property.

[0019] The deposit information of Lactiplantibacillus plantarum Lp116 of the present invention is as follows:

[0020] Deposit number: CCTCC NO:M 20241784;

[0021] Classification and nomenclature: Lactiplantibacillus plantarum Lp116;

[0022] Depository: China Center for Type Culture Collection;

[0023] Address of depository: Wuhan University, Wuhan, China;

[0024] Date of preservation: August 14, 2024. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a graph showing the effect of Lactobacillus plantarum Lp116 of Experimental Example 5 of the present invention on the food intake and water intake of type 2 diabetic mice.

[0026] Figure 2 This is a graph showing the effect of Lactobacillus plantarum Lp116 on the body weight of type 2 diabetic mice in Experimental Example 5 of the present invention.

[0027] Figure 3 This is a graph showing the effect of Lactobacillus plantarum Lp116 of Experimental Example 6 of the present invention on the blood glucose level of type 2 diabetic mice.

[0028] Figure 4 This is a graph showing the effect of Lactobacillus plantarum Lp116 of Experimental Example 6 of the present invention on oral glucose tolerance in type 2 diabetic mice.

[0029] Figure 5 This is a graph showing the effects of Lactobacillus plantarum Lp116 on type 2 diabetic mice and insulin resistance in Experimental Example 6 of the present invention.

[0030] Figure 6 This is a graph showing the effect of Lactobacillus plantarum Lp116 of Experimental Example 8 of the present invention on inflammatory factors in type 2 diabetic mice.

[0031] Figure 7 This is the Chao1 index and Simpson index graph of mouse intestinal microorganisms in Experimental Example 9 of the present invention.

[0032] Figure 8 This is a β-diversity analysis diagram of the mouse intestinal flora in Experimental Example 9 of the present invention.

[0033] Fig. 9 This is a bar chart of the abundance of various types of bacterial flora in the mouse intestine of Experimental Example 9 of the present invention.

[0034] The markings in the figure are listed as follows:

[0035] Figure 1-Figure 9The group numbers are as follows: NC is the normal group mice, MC is the model group mice, MET is the drug treatment group mice, and Lp116 is the probiotic group mice.

[0036] Figure 3 The vertical axis FBG in the right figure refers to the fasting blood glucose value.

[0037] Figure 4 The vertical axis OGTT AUC in the right graph refers to the value of the area under the curve (AUC) of the oral glucose tolerance test (OGTT).

[0038] Figure 5 The vertical axis HOMA-IR refers to the insulin resistance index.

[0039] Figure 8 The horizontal and vertical axes of NMDS1 refer to non-metric multidimensional scaling. DETAILED DESCRIPTION

[0040] The specific contents of the present invention are further described in detail below in conjunction with specific embodiments, experimental examples and drawings, but the protection scope of the present invention is not limited thereto.

[0041] Sources of biological materials

[0042] 1. The Escherichia coli, Staphylococcus aureus and Salmonella used in Experimental Example 3 were purchased from Guangdong Microbiological Culture Collection Center. Streptococcus pharyngitis was Streptococcus pharyngitis strain 4997, which has been registered in NCBI with the accession number MT459365; Streptococcus pseudopneumoniae was Streptococcus pseudopneumoniae strain 5027, which has been registered in NCBI with the accession number MT512127.

[0043] 2. The mice used in Experimental Examples 5-9 are commercially available.

[0044] “Above” and “below” in this article include the original number.

[0045] The “Lactiplantibacillus plantarum strain Lp116”, “Lactiplantibacillus plantarum strain Lp116”, “Lactiplantibacillus plantarum Lp116”, “Lactiplantibacillus plantarum Lp116”, “Lp116”, “Lp116 strain”, and “strain Lp116” in this article all refer to: Lactiplantibacillus plantarum strain Lp116 with a preservation number of CCTCC NO: M 20241784.

[0046] Group 1 Example, Lp116 strain of the present invention

[0047] This group of embodiments provides a plant lactobacillus (Lactiplantibacillus plantarum) Lp116 with blood sugar lowering ability. All embodiments in this group have the following common features: the deposit number of the plant lactobacillus (Lactiplantibacillus plantarum) Lp116 with blood sugar lowering ability is CCTCC NO: M 20241784.

[0048] Any act of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, or promising to sell a strain of Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784, and / or using a strain of Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 in combination with other probiotics, and / or using a strain of Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 to antagonize various pathogens, and / or preparing antibacterial products, and / or preparing drugs for improving constipation or regulating intestinal flora, and / or preparing health foods with laxative function or intestinal flora regulating function all fall within the protection scope of the present invention.

[0049] The other probiotics include but are not limited to: Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii lactic acid subsp., Lactobacillus helveticus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Saccharomyces cerevisiae, Delb's spore-forming torula, Candida, Wickham's yeast, Pichia, Saccharomyces boulardii, Torulopsis spp., Saccharomyces cerevisiae, Rhodotorula, Schizosaccharomyces pombe , Saccharomyces cerevisiae, Bacillus thuringiensis, Bacillus laterosporus, Bacillus megaterium, Bacillus mucilaginosus, Bacillus azotobacterium, Bacillus sphaericus, Clostridium butyricum, Bifidobacterium adolescentis, Bifidobacterium angularis, Bifidobacterium animalis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium bovis, Bifidobacterium breve, Bifidobacterium dentate, Bifidobacterium infantis (i.e., Bifidobacterium longum infantis subsp. infantis), Bifidobacterium lactis (i.e., Bifidobacterium animalis subsp. cremoris), Bifidobacterium longum, Bifidobacterium pseudocatenulatum, as well as Bifidobacterium thermophilum and Bifidobacterium acidothermophilum.

[0050] Those skilled in the art can make conventional selections or adjustments to pharmaceutical excipients according to actual production needs, in combination with conventional technical means or basic common sense of pharmaceutical production processes (for example, "Encyclopedia of Preparation Technology", "Pharmaceutical Preparation Technology", etc.), and then prepare a strain of Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 into products with different dosage forms, different storage conditions, and different shelf lives. This is possible and easy to do without technical barriers for those skilled in the art.

[0051] The second group of examples, use of the Lp116 strain of the present invention

[0052] This group of embodiments provides the use of Lactobacillus plantarum Lp116 with a deposit number of CCTCC NO: M 20241784 in preparing hypoglycemic drugs or health foods with auxiliary hypoglycemic function or antibacterial agents or non-therapeutic antibacterial agents.

[0053] In a specific embodiment, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis, and Streptococcus pseudopneumoniae.

[0054] In some embodiments, the glucose-lowering drug uses Lactobacillus plantarum Lp116 as the active ingredient.

[0055] The third group of examples, the hypoglycemic agent of the present invention

[0056] This group of embodiments provides a hypoglycemic drug. All embodiments in this group have the following common features: the hypoglycemic drug comprises a pharmacologically active ingredient, and the pharmacologically active ingredient comprises: Lactobacillus plantarum Lp116 with a deposit number of CCTCC NO: M 20241784.

[0057] In a further embodiment, the glucose-lowering drug further comprises: a pharmaceutically acceptable excipient.

[0058] In a more specific embodiment, the pharmaceutically acceptable excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0059] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned pharmaceutically acceptable excipients, and prepare Lactobacillus plantarum Lp116 with a deposit number of CCTCC NO: M 20241784 into different dosage forms, such as powders, tablets, gels, sprays, granules, capsules, liquids, etc.

[0060] In a specific embodiment, the dosage form of the drug is selected from: one or more of powder, tablet, gel, spray, granule, capsule, liquid.

[0061] The fourth group of embodiments, the health food with auxiliary hypoglycemic function of the present invention

[0062] This group of embodiments provides a health food with auxiliary blood sugar lowering function. All embodiments in this group have the following common features: the health food with auxiliary blood sugar lowering function comprises: Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M20241784.

[0063] In a further embodiment, the aforementioned health food with auxiliary blood sugar lowering function further includes: auxiliary materials.

[0064] In a more specific embodiment, the excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0065] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Lactobacillus plantarum Lp116 with a deposit number of CCTCC NO: M20241784 into different dosage forms, such as powders, tablets, gels, sprays, granules, capsules, liquids, etc.

[0066] In a specific embodiment, the dosage form of the product is selected from: one or more of powder, tablet, gel, spray, granule, capsule, liquid.

[0067] The fifth group of embodiments, the antibacterial method of the present invention

[0068] This group of embodiments provides a non-therapeutic antibacterial method. All embodiments in this group have the following common features: Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 is used for antibacterial.

[0069] In a specific embodiment, the antibacterial activity refers to: inhibiting Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis, and Streptococcus pseudopneumoniae.

[0070] Experimental Example 1: Isolation, screening and identification of Lactobacillus plantarum

[0071] (1) Isolation and screening of Lactobacillus plantarum

[0072] Collect fecal samples from infants, take 0.5g to 4.5mL sterile saline, shake and disperse the sample thoroughly, take 100μL sample for gradient dilution, select appropriate gradient and evenly spread on MRS agar plate, place under anaerobic conditions, culture at 37℃ for 36-48h, select single colonies, perform Gram staining, observe the morphological characteristics of colonies under a microscope, and preliminarily screen suspected lactobacillus strains. The strains were repeatedly streaked and purified on MRS solid medium to obtain purified lactobacillus strains, which were stored in -80℃ glycerol tubes.

[0073] MRS medium formula: peptone 10g, beef extract powder 5g, yeast powder 5g, glucose 20g, anhydrous sodium acetate 5g, diammonium hydrogen citrate 2g, Tween 80 1mL, K2HPO4 2g, MgSO4 0.2g, MnSO40.05g, 1000ml distilled water, pH 6.5, high pressure sterilization at 121℃ for 20min, add 1.5g agar powder to the solid MRS medium.

[0074] (2) Identification and preservation of Lp116 strains

[0075] The selected target strains were cultured, the cells were collected, the genomic DNA was extracted, and the universal primers 27F and 1492R recorded in paragraph 58 of Chinese invention patent ZL202210478937.4 were used to amplify the 16SrDNA fragment, the PCR amplification products were detected by agarose gel electrophoresis, and the PCR products were sequenced. The PCR reaction system: 10×buffer 10μL, 10mMdNTP 2μL, 1μL of upper and lower primers, 2μL of DNA template, 0.5μL of Taq enzyme, 34.5μL of ddH2O. Pre-denaturation at 95℃ for 10min; then 94℃30s, 60℃30s, 72℃1min for a total of 35 cycles, and then extended at 72℃ for 5min. The PCR product was detected by gel electrophoresis and sent to Wuhan Jinkairui Bioengineering Co., Ltd. for sequencing. The identified gene sequence was compared in the NCBI database using the BLAST tool. Based on the molecular biological identification results, the strain was determined to be Lactobacillus plantarum. The strain was named Lactobacillus plantarum Lp116 and deposited. The deposit information is as follows:

[0076] Deposit number: CCTCC NO:M 20241784;

[0077] Classification and nomenclature: Lactiplantibacillus plantarum Lp116;

[0078] Depository: China Center for Type Culture Collection;

[0079] Address of depository: Wuhan University, Wuhan, China;

[0080] Date of preservation: August 14, 2024.

[0081] Experimental Example 2: Gastric Juice and Bile Salt Tolerance Experiment

[0082] Simulated artificial gastric juice: prepare 0.5% sodium chloride solution, adjust the pH to 2.5 with 1 mol / L HCl, add 0.3% pepsin, and filter with a 0.22 μm microporous filter membrane to sterilize after fully dissolving.

[0083] Bile salt solution: Weigh 0.2% sodium thioglycolate and 0.3% porcine bile salt and add them to L-MRS liquid culture medium. After fully dissolving, sterilize in an autoclave at 115°C for 20 min for later use.

[0084] After the strain to be tested is activated for 2 generations, the concentration of the bacterial solution is adjusted to 10 8CFU / mL. Take 1mL of bacterial suspension and centrifuge it, collect the bacteria, inoculate them into 1mL of prepared artificial gastric juice and bile salt solution, mix them, digest them at 37℃, and take the digestion fluid at 0h and 3h to detect the number of viable bacteria, calculate the survival rate, and the results are shown in Table 1. Among them, strain survival rate (%) = Nt / N0×100%, where N0 represents the number of viable bacteria of the strain at 0h, lgCFU / mL, and Nt represents the number of viable bacteria of the strain at 3h, lgCFU / mL.

[0085] Table 1. Survival rate of Lactobacillus plantarum Lp116 tolerant to artificial gastric juice and bile salt solution

[0086] strain Artificial gastric juice (%) Bile salts (%) Lactobacillus plantarum Lp116 96.85±0.20％ 92.53±0.10％

[0087] The experimental results showed that after 3 hours of gastric juice digestion, the survival rate of Lactobacillus plantarum Lp116 was 96.85%, and after 3 hours of bile salt digestion, the survival rate of Lactobacillus plantarum Lp116 was 92.53%, which indicated that Lactobacillus plantarum Lp116 had good resistance to gastric acid and bile salt.

[0088] Experimental Example 3: Inhibitory effect of Lactobacillus plantarum on pathogenic bacteria

[0089] The MRS broth containing 1.5% agar was cooled to about 55°C and mixed with the suspensions of Escherichia coli, Salmonella, Staphylococcus aureus, Streptococcus pharyngitis and Streptococcus pseudopneumoniae in a certain proportion to make the number of live indicator bacteria reach 10. 6 CFU / mL, and then quickly poured into the plate with the Oxford cup placed in advance. After the culture medium cooled and solidified, the Oxford cup was taken out, and 200 μL of Lactobacillus plantarum Lp116 fermentation liquid was injected into each well. After overnight culture at 37°C, the diameter of the inhibition zone was measured.

[0090] Table 2. Diameter of inhibition zone of Lactobacillus plantarum against pathogens (mm)

[0091]

[0092]

[0093] It can be seen from the above table that Lactobacillus plantarum can effectively inhibit intestinal pathogens, especially Staphylococcus aureus, with an inhibition zone diameter of 27.33 mm, showing good inhibition ability.

[0094] Experimental Example 4: Inhibition of α-glucosidase by Lactobacillus plantarum

[0095] Lactobacillus plantarum was inoculated into liquid L-MRS medium and cultured overnight at 37°C. After the second generation, the culture was cultured for 8 hours. After that, the third generation was cultured for 14 hours. The fermentation liquid was centrifuged, the supernatant was discarded, and the liquid was resuspended with physiological saline. The concentration of the bacterial liquid was adjusted to 108 Reserve at CFU / mL. Add 150 μL of PBS solution, 75 μL of p-nitrophenyl-α-D-glucopyranoside (PNPG) solution and 25 μL of the sample to be tested. Place the mixture on ice for 3 min, add 50 μL of α-glucosidase solution at 0.2 U / mL, react in a water bath at 37 °C for 15 min, add 1 mL of 1 mol / L Na2CO3 as the reaction termination solution, and measure the absorbance of the reaction solution at 405 nm:

[0096] A: Add sample and α-glucosidase;

[0097] B: Add sample without adding α-glucosidase;

[0098] C: Do not add sample but add α-glucosidase;

[0099] D: Do not add sample and do not add α-glucosidase

[0100] Table 3. Inhibition rate of Lactiplantibacillus plantarum Lp116 on α-glucosidase

[0101] strain α-Glucosidase inhibition rate (%) Lactobacillus plantarum Lp116 16.00±1.00

[0102] As can be seen from the above table, Lactiplantibacillus plantarum has the ability to inhibit α-glucosidase, indicating that the strain has potential hypoglycemic ability.

[0103] Experimental example 5. Effects of Lactiplantibacillus plantarum on food intake, water intake and body weight of type 2 diabetic mice

[0104] 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 license number for the use of experimental animals is: SCXK(E)2020 - 0019. The animal experiment was maintained at 20 - 22 °C, humidity 40 - 60%, with a 12 h light / 12 h dark cycle. Mice had free access to food and water. After one week of adaptation, according to the reference F. Yan, N. Li, J. Shi, H. Li, Y. Yue, W. Jiao, N. Wang, Y. Song, G. Huo and B. Li, Lactiplantibacillus plantarum alleviates type 2 diabetes by regulating hepatic glucose and lipid metabolism and gut microbiota in mice, Food Funct, 2019, the mice were induced to develop type 2 diabetes. The mice were divided into 4 groups (n = 12): normal group (NC group), model group (MC group), probiotic group (Lp116, 6*10 9CFU / d), and drug treatment group (MET, 0.2g / kg / d). The normal group and model group were gavaged with sterile saline, the probiotic group was gavaged with Lactobacillus plantarum Lp116 once a day for 28 consecutive days, and the drug treatment group was gavaged with metformin hydrochloride (MET) once a day for 28 consecutive days. The daily food intake, daily water intake, and weekly body weight of the mice were monitored.

[0105] The typical characteristics of type 2 diabetic mice are mainly manifested in increased daily food intake, increased water intake, increased urine output, and decreased body weight. After four weeks of intragastric administration of probiotics, the overall symptoms of diabetes were alleviated and improved. Figure 1 The treatment of the strain effectively controlled the increase in the amount of food and water consumed by the mice, achieving significant improvement compared with the model group. Daily monitoring of the humidity of the litter showed that the urine volume in the cages of the treatment group had a trend of returning to normal. Figure 2 The changes in the weight values ​​of mice indicate that the intervention of probiotics has a good stabilizing or restoring effect on the decreased weight values ​​of mice, and effectively improves the absorption of nutrients by type 2 diabetic mice, which overall show a trend of approaching the healthy group.

[0106] Experimental Example 6: Effect of Lactobacillus plantarum on glucose metabolism in type 2 diabetic mice

[0107] Before treatment and during the four-week treatment period, blood samples were collected from the tail vein of the mice every week to measure the fasting blood glucose level. The fasting blood glucose level (FBG) measurement results are shown in Figure 2. Figure 3 As shown. After five weeks of treatment, the oral glucose tolerance of mice was tested. Each group of mice was given oral glucose, and the blood glucose level was measured at 0min, 15min, 30min, 60min, 90min, and 120min after oral glucose tolerance test. The area below the oral glucose tolerance line in the oral glucose tolerance test was measured. At the end of the experiment, serum was taken to measure the serum insulin concentration of mice.

[0108] Experimental steps for oral glucose tolerance: The amount of glucose used in the mouse glucose tolerance experiment is 2g per kilogram of body weight (2g / kg). Prepare a glucose solution of appropriate concentration according to the experimental requirements, and use normal saline to prepare a 20% glucose solution. The mice were fasted for 16 hours, during which time the mice maintained normal drinking water; before starting the glucose tolerance experiment, weigh each mouse. Blood was collected from the tip of the mouse's tail, and the fasting blood sugar was measured with a blood glucose meter, and the measured value was considered the blood sugar value at 0min. Glucose was gavaged and the timing began from the moment it was completed. The blood sugar value of each mouse was measured at 15, 30, 60, 90 and 120min.

[0109] The results of the weekly blood glucose test of mice are as follows Figure 3As shown, it can be concluded that the strain can significantly reduce the blood sugar level of diabetic mice after four weeks of intervention. As the treatment time increases, the blood sugar lowering effect of mice becomes more obvious. In the fourth week of treatment, the fasting blood sugar levels of mice in the probiotic group and the model group reached a very significant difference, indicating that the Lp116 strain can effectively reduce the blood sugar level of mice and bring it closer to the healthy group. At the end of the four-week treatment, the oral glucose tolerance index of the mice was measured. Figure 4 As shown, the area below the oral glucose tolerance line of the Lp116 treatment group reached a very significant level compared with the model group. The experimental results showed that the Lp116 strain not only lowered the blood sugar of mice, but also had a good effect on improving the glucose absorption and metabolism function of diabetic mice.

[0110] The insulin resistance index (HOMA-IR) is a value calculated based on the serum insulin level and fasting blood glucose value. It is used to determine the degree of insulin resistance. Figure 5 It can be concluded that the insulin resistance of the Lp116 strain treatment group and the drug treatment group was significantly improved and alleviated, and was close to that of the healthy group.

[0111] The above results indicate that the treatment with Lp116 strain can improve and enhance the glucose metabolism ability of mice, thereby lowering the blood sugar levels of mice and thus controlling the progression of the disease in mice.

[0112] Experimental Example 7: Effect of Lactobacillus plantarum on blood lipid levels in type 2 diabetic mice

[0113] The indicators of mouse serum were measured: triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), as shown in Table 4:

[0114] Table 4. Results of total cholesterol, total triglycerides, high-density lipoprotein, and low-density lipoprotein in serum

[0115] Group TG (mmol / L) TC(mmol / L) HDL-C (mmol / L) LDL-C (mmol / L) Lp116 group 1.13±0.14b 8.75±0.99a 1.95±0.33a 3.19±0.30a MET Group 1.37±0.29ab 6.91±0.48a 1.54±0.17a 2.70±0.56a MC Group 1.69±0.30a 9.95±1.05a 2.22±0.11a 3.21±0.25a NC Group 0.87±0.23c 5.26±0.63b 1.60±0.27a 2.46±0.30a

[0116] As shown in Table 4, oral administration of Lp116 strain has a certain effect on reducing the blood lipid level of type 2 diabetic mice, and it is close to the healthy group. The indicators of high-density lipoprotein and total cholesterol showed a positive therapeutic effect, and the total triglyceride concentration of the Lp116 strain treatment group returned to the level of the healthy group (NC) and was within the normal range. There was a significant difference between the Lp116 strain and the model group (MC). In the treatment of diabetic dyslipidemia, the Lp116 strain is particularly effective in reducing serum triglycerides.

[0117] The above results indicate that the Lp116 strain can effectively reduce the blood lipid levels of type 2 diabetic mice and has a very good effect.

[0118] Experimental Example 8: Effects of Lactobacillus plantarum on inflammatory factors in type Ⅱ diabetic mice

[0119] The onset of diabetes will cause systemic inflammation in the body. The occurrence of chronic inflammatory diseases will lead to an increase in tumor necrosis factor in the patient's serum and promote the production of interleukin 6. At the end of the experiment, the whole blood of the mice was centrifuged to obtain serum, and the ELISA kit was used to measure the inflammatory factors in the mouse serum, including the concentration levels of LPS (lipopolysaccharide), IL-6, TNF-α-, and Insulin. Figure 6 As shown in a, 6c, and 6d, compared with the model group (MC), oral administration of Lp116 strains reduced inflammation in mice and slowed down the occurrence of inflammation in the body. Serum insulin level is usually an important indicator for diagnosing type 2 diabetes and is also an important monitoring indicator during treatment. During the onset of the disease, the insulin concentration in the patient's serum will be high, such as Figure 6 As shown in b, oral administration of Lp116 effectively reduced the serum insulin level of mice and brought it closer to the level of the healthy group.

[0120] The above data results show that oral administration of Lp116 strain has a good effect in reducing systemic inflammation in type 2 diabetic mice.

[0121] Experimental Example 9: Effect of Lactobacillus plantarum on intestinal flora of type 2 diabetic mice

[0122] After the experiment, the mice were killed, the abdomen was disinfected and dissected, and the cecal contents of each group of mice were taken for microbial diversity analysis. The 16S rDNA V3-V4 hypervariable regions of all bacteria in the samples were sequenced on the Mi Seq Illumina sequencing platform to determine the characteristics of the intestinal microbiota.

[0123] α-diversity reflects the species richness and diversity of the sample. Chao1 index and Shannon index are commonly used indicators for judging the diversity and abundance of intestinal microbial colonies. Figure 7 As shown in the figure, the abundance and diversity of intestinal flora in type 2 diabetic mice were significantly decreased compared with those in the healthy group (p<0.01). According to the Chao1 index analysis, the treatment with Lp116 strain can increase and improve the abundance of intestinal flora, which is lower than that in the model group but reaches a significant difference. The effect is equivalent to that in the drug treatment group. According to the Simpson index analysis, oral administration of Lp116 strain can significantly increase the diversity of intestinal flora in type 2 diabetic mice and approach that of the healthy group. The results show that Lp116 strain can increase and improve the abundance and diversity of intestinal flora in type 2 diabetic mice.

[0124] β-diversity is used to compare the similarity of different samples in terms of species diversity. NMDS is a commonly used analysis method that can better reflect the nonlinear structure of ecological data. The closer the distance on the coordinate graph, the higher the similarity. Figure 8 As shown, the two sides of the coordinate axis are the model group and the healthy control group, and the drug treatment group and the strain treatment group are between the two groups and gradually approach the healthy group, indicating that the strain effectively improves the intestinal flora of type 2 diabetic mice and tends to be normal.

[0125] Metformin is a first-line drug for the treatment of type 2 diabetes, and its mechanism of action is constantly being revealed by research. Many studies have shown that metformin can have an effect on type 2 diabetic mice by changing the intestinal flora. Fig. 9 Results showed that Oscillospira was significantly increased in the intestinal flora of mice gavaged with Lp116. According to literature reports, Oscillospira belongs to the Firmicutes phylum and can produce short-chain fatty acids such as butyrate, which is closely related to human health. The increase in the abundance of Oscillospira can affect serum metabolites and help regulate glucose homeostasis and blood lipids. In addition, the abundance of beneficial intestinal bacteria such as Blautia, Colidextribacter, unclassified lachnospiraceae, and unclassified Bacteroides increased significantly in the Lp116 treatment group (p<0.01). At the level of Akkermansia, the abundance of the treatment group was significantly higher than that of drug treatment. Harmful intestinal bacteria such as Bacteroides, Helicobacter, Desulfovibrio, and Escherichia showed a significant decrease in the Lp116 treatment group and the drug treatment group (p<0.05).

[0126] The above results show that type 2 diabetic mice have intestinal flora disorders, and the treatment of Lactobacillus plantarum Lp116 can effectively reduce the abundance of harmful intestinal flora in type 2 diabetic mice and significantly increase the abundance of beneficial intestinal flora. And by improving the species diversity and abundance of intestinal flora as a whole, the blood sugar level of type 2 diabetic mice can be effectively controlled and improved.

Claims

1. A Lactobacillus plantarum Lp116 having the ability to lower blood sugar, characterized in that: Its deposit number is CCTCC NO:M 20241784.

2. Use of Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 in preparing hypoglycemic drugs or health foods with auxiliary hypoglycemic function or antibacterial agents or antibacterial agents for non-therapeutic purposes.

3. The use of Lactobacillus plantarum Lp116 with the deposit number of CCTCC NO: M 20241784 according to claim 2 for preparing hypoglycemic drugs or health foods with auxiliary hypoglycemic function or antibacterial agents or non-therapeutic antibacterial agents, characterized in that: The antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis and Streptococcus pseudopneumoniae.

4. The use of Lactobacillus plantarum Lp116 with the deposit number of CCTCC NO: M 20241784 according to claim 2 or 3 for preparing hypoglycemic drugs or health foods with auxiliary hypoglycemic function or antibacterial agents or non-therapeutic antibacterial agents, characterized in that: The hypoglycemic drug uses Lactobacillus plantarum Lp116 as a pharmaceutically active ingredient.

5. A hypoglycemic drug comprising a pharmacologically active ingredient, characterized in that: The pharmacologically active ingredients include: Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784.

6. A hypoglycemic agent according to claim 5, characterized in that: Also includes: Pharmaceutically acceptable excipients.

7. A health food with auxiliary blood sugar lowering function, characterized in that: include: Lactobacillus plantarum Lp116 with the deposit number of CCTCCNO:M 20241784.

8. The blood sugar lowering health food according to claim 7, characterized in that: Also includes: Auxiliary materials.

9. A non-therapeutic antibacterial method, characterized in that: Lactobacillus plantarum Lp116 with a preservation number of CCTCC NO: M 20241784 was used for bacteriostasis.

10. A non-therapeutic destination antibacterial method according to claim 9, characterized in that: The antibacterial agent refers to: inhibiting Escherichia coli, Staphylococcus aureus, Salmonella, Streptococcus pharyngitis, and Streptococcus pseudopneumoniae.

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