Lactobacillus acidophilus and application thereof in preparation of medicine for preventing and / or treating hyperlipidemia and hyperglycemia

Abstract

The invention relates to the field of microorganisms, and discloses a strain of lactobacillus acidophilus and application of the lactobacillus acidophilus in preparation of drugs for preventing and / or treating hyperlipidemia and hyperglycemia, the preservation number of the lactobacillus acidophilus is CGMCC No.33155, the strain can generate lactic acid, is resistant to acid and cholate, and has strong self-aggregation ability and hydrophobic ability. The strain provided by the invention can promote the decomposition of fat and sugar by degrading cholesterol and inhibiting the activity of alpha-glucosidase and alpha-amylase, can reduce the expression levels of blood sugar, blood fat and inflammatory factors of hyperlipidemia and hyperglycemia mice, and provides a new idea for preparing medicines for preventing and / or treating hyperglycemia and hyperlipidemia.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to a strain of Lactobacillus acidophilus and application thereof in preparing a medicine for preventing and / or treating hyperlipidemia and hyperglycemia. Background Art

[0002] As people's living standards improve, high-sugar and high-fat diets have become commonplace. However, due to irregular and uncontrolled diets, cardiovascular diseases such as high blood sugar, high blood lipids and high blood pressure follow. It is reported that the mortality rate of cardiovascular diseases ranks first, higher than that of tumors and other diseases. Nowadays, the group of people with "three highs" diseases has gradually become younger, and the prevention and treatment of "three highs" diseases is imminent.

[0003] Current research has confirmed that probiotics have significant effects in alleviating and preventing chronic diseases such as hyperlipidemia and hyperglycemia. Compared with traditional drug treatments, probiotics have no side effects on the body and do not cause complications. They can also produce beneficial effects on the human body by regulating the intestinal flora and producing short-chain fatty acids. However, existing microorganisms have disadvantages such as unstable viable counts and cannot stably function in the human body.

[0004] Therefore, there is an urgent need to breed strains with excellent fermentation characteristics, high viable bacterial counts, certain probiotic functions, and at the same time the ability to survive stably in the human body and to prepare drugs for the prevention and / or treatment of hyperlipidemia and hyperglycemia. Summary of the invention

[0005] The purpose of the present invention is to overcome the problems existing in the prior art and to provide a strain of Lactobacillus acidophilus and its use in preparing a drug for preventing and / or treating hyperlipidemia and hyperglycemia.

[0006] The first aspect of the present invention provides a strain of Lactobacillus acidophilus 嗜酸乳杆菌 The preservation number of the Lactobacillus acidophilus is CGMCC No.33155.

[0007] A second aspect of the present invention provides a bacterial agent, wherein the bacterial agent contains the Lactobacillus acidophilus as described above.

[0008] The third aspect of the present invention provides the use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for decomposing cholesterol.

[0009] The fourth aspect of the present invention provides the use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for inhibiting the activity of α-glucosidase.

[0010] A fifth aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for preventing and / or treating hyperlipidemia.

[0011] A sixth aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for preventing and / or treating hyperglycemia.

[0012] A seventh aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for reducing inflammatory factors.

[0013] Through the above technical solution, the beneficial effects of the present invention at least include: The Lactobacillus acidophilus of the present invention is resistant to gastric acid and bile salts, can decompose carbohydrates into high lactic acid, has high α-glucosidase and α-amylase inhibition rates and high cholesterol clearance rates, can reduce blood sugar, blood lipids and inflammatory factors in hyperlipidemia and hyperglycemia mice, and has the prospect of preparing drugs for preventing and / or treating hyperglycemia and hyperlipidemia.

[0014] Biological Deposit The strain provided by the present invention is classified and named as Lactobacillus acidophilus 嗜酸乳杆菌 , was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration (abbreviated as CGMCC) on December 23, 2024. Its deposit number is CGMCC No.33155, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0015] The strain provided by the present invention is classified and named as Lactobacillus plantarum 植物乳杆菌 , was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration (abbreviated as CGMCC) on December 23, 2024. Its deposit number is CGMCC No.33152, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is a morphological diagram of a Lactobacillus acidophilus colony provided by the present invention; Figure 2 It is a Gram microscopic image of a Lactobacillus acidophilus colony provided by the present invention. DETAILED DESCRIPTION

[0017] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0018] The inventor of the present invention accidentally isolated a strain of Lactobacillus acidophilus. Figure 1 and Figure 2As shown, the colony of the strain is dot-shaped, white and opaque, full in shape, smooth and moist, and the bacterial lawn grows along the streaks. It is determined to be Lactobacillus acidophilus and named Lactobacillus acidophilus CCNH655.

[0019] Based on the above findings, the present invention provides a strain of Lactobacillus acidophilus in a first aspect. 乳杆菌属 嗜酸的 The preservation number of the Lactobacillus acidophilus is CGMCC No.33155.

[0020] A second aspect of the present invention provides a bacterial agent, wherein the bacterial agent contains the Lactobacillus acidophilus as described above.

[0021] The present invention has no particular limitation on the type of the bacterial agent, which can be any type of bacterial agent currently available in the art. According to a preferred embodiment of the present invention, the bacterial agent is selected from at least one of a liquid bacterial agent, a concentrated bacterial agent and a solid bacterial agent.

[0022] In the present invention, the bacterial agent may further contain auxiliary materials, which may be protective agents (e.g., freeze-dried protective agents, such as skim milk powder, maltodextrin, trehalose, dextran, corn oil and glycerol, etc.) and / or buffers (e.g., buffers remaining in the process of preparing solid bacterial agents, etc.). Preferably, the auxiliary materials may be selected from glycerol and / or corn oil.

[0023] In the present invention, the number of viable Lactobacillus acidophilus in the bacterial agent may be no less than 10 4 CFU / mL.

[0024] The third aspect of the present invention provides the use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for decomposing cholesterol.

[0025] Preferably, the cholesterol is glycocholic acid and / or taurocholic acid.

[0026] The fourth aspect of the present invention provides the use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for inhibiting the activity of α-glucosidase.

[0027] A fifth aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for preventing and / or treating hyperlipidemia.

[0028] A sixth aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for preventing and / or treating hyperglycemia.

[0029] According to the present invention, the hyperglycemia is caused by type 2 diabetes.

[0030] According to one embodiment of the present invention, the strain provided by the present invention can improve the physiological indicators of mice already suffering from diabetes, so the strain provided by the present invention can be used in the preparation of drugs for treating hyperglycemia caused by diabetes.

[0031] A seventh aspect of the present invention provides use of the aforementioned Lactobacillus acidophilus in the preparation of a medicament for reducing inflammatory factors.

[0032] Preferably, the inflammatory factors include tumor necrosis factor-α (TNF-α) and / or interleukin-6 (IL-6).

[0033] In the present invention, the drug may contain pharmaceutically acceptable excipients, including corn oil and / or glycerol.

[0034] The present invention also provides the use of the aforementioned Lactobacillus plantarum in the preparation of a medicine for inhibiting the reproduction of pathogenic bacteria.

[0035] According to the present invention, the pathogenic bacteria may include Escherichia coli ( 大肠杆菌 ), Staphylococcus aureus ( 金黄色葡萄球菌 ),salmonella( 沙门氏菌属 ) and Candida albicans ( 白色念珠菌 ) at least one of the following.

[0036] According to some embodiments of the present invention, the Salmonella preferably includes Salmonella typhimurium ( 鼠伤寒沙门氏菌 ) and Salmonella Enteritidis ( 肠炎沙门氏菌 ).

[0037] The present invention also provides a composite bacterial agent, which comprises Lactobacillus acidophilus with a preservation number of CGMCC No. 33155 and Lactobacillus plantarum with a preservation number of CCGMCC No. 33152 provided by the present invention. 乳杆菌属 植物的 .

[0038] As shown in the application document with application number 202510222543.6, Lactobacillus plantarum with a deposit number of CCGMCC No. 33152 has the effects of antibacterial, inhibiting the reproduction of pathogens and reducing the expression of pro-inflammatory factors, and also has biosafety and antibiotic sensitivity. Therefore, when Lactobacillus plantarum with a deposit number of CCGMCC No. 33152 is used to prepare a drug for preventing and / or treating vaginitis, it can be used in combination with a drug prepared by Lactobacillus acidophilus with a deposit number of CGMCC No. 33155 provided by the present application to further enhance the antibacterial and anti-inflammatory effects, that is, Lactobacillus plantarum with a deposit number of CCGMCC No. 33152 and Lactobacillus acidophilus provided by the present application can be used to compound a composite bacterial agent to prepare a drug for preventing and / or treating vaginitis.

[0039] In the composite bacterial agent provided by the present invention, the two strains can be packaged independently or mixed, but in order to maintain the activity of the strains to the greatest extent to exert the effect, they are preferably packaged independently.

[0040] When a patient uses a medicine containing the composite bacterial agent provided by the present invention, the medicine containing the two strains can be used simultaneously or sequentially. However, in order to maximize the effect, it is preferably used sequentially, but the order of sequential use is not limited.

[0041] The present invention will be described in detail below through examples.

[0042] In the following examples, unless otherwise specified, the chemical / biological reagents used were purchased from regular chemical / biological reagent suppliers.

[0043] Lactobacillus rhamnosus ( 鼠李糖乳杆菌 )LGG, number ATCC 53103, purchased from the American Type Culture Collection; MRS liquid culture medium was purchased from Qingdao Haibo Biological, catalog number HB0384-1; PBS buffer was purchased from Solebol, catalog number P1003; Escherichia coli ( 大肠杆菌 ), numbered (1) ATCC 25922, purchased from the American Type Culture Collection, (2) CICC 10421, purchased from the China Industrial Microbiological Culture Collection; Staphylococcus aureus ( 金黄色葡萄球菌 ), numbered CMCC(B)26001 and CMCC(B)26003, both purchased from the Chinese Medical Bacteria Collection Center; Salmonella, (1) Salmonella typhimurium ( 鼠伤寒沙门氏菌 ), numbered ATCC 14028, purchased from the American Type Culture Collection; (2) Salmonella Enteritidis ( 肠炎沙门氏菌 ), numbered CVCC 3378, purchased from the National Veterinary Microbiological Bacteria (Virus) Collection Center; Candida albicans ( 白色念珠菌 ), with the numbers (1) CICC 1965, purchased from China Industrial Microbiological Culture Collection, and (2) ATCC 10231, purchased from American Type Culture Collection.

[0044] Example 1 The strains of Lactobacillus acidophilus CCNH655 and LGG glycerol tube were inoculated into fresh MRS liquid medium at 2 vol%, cultured at 37°C overnight for 18 h, and the OD of the two strains was measured. 600 value, add MRS liquid medium to the culture medium, so that the OD 600 The values ​​were equal to obtain an activated bacterial solution, which was inoculated into fresh MRS liquid culture medium at an inoculum size of 2 vol%, and cultured at 37°C, 200 rpm for 18 h to obtain the expanded culture solution of Lactobacillus acidophilus CCNH655 and the expanded culture solution of LGG.

[0045] Measure the OD of Lactobacillus acidophilus CCNH655 and LGG culture solutions 600 They were 8.71 and 8.13 respectively. The growth performance of Lactobacillus acidophilus CCNH655 was better than that of LGG.

[0046] Example 2 Take the culture medium of Lactobacillus acidophilus CCNH655 and LGG, and refer to T / CNHFA435-2024 "Test Method for Gastric Juice Tolerance of Probiotics" to evaluate the strains' resistance to gastric acid and bile salts.

[0047] In the gastric acid resistance test, the pH was 3 and the incubation was 2 h; in the bile salt resistance test, the bile salt content was 0.2 wt % and the incubation was 4 h.

[0048] The survival rates of Lactobacillus acidophilus CCNH655 and LGG in the gastric acid resistance experiment were 95.48% and 93.33%, respectively, and the survival rates in the bile salt resistance experiment were 75.21% and 53.13%, respectively. It can be found that the gastric acid resistance and bile salt resistance of Lactobacillus acidophilus CCNH655 and LGG are better than those of LGG.

[0049] Example 3 600 μL of the activated bacterial solution was inoculated into 30 mL of fresh MRS liquid medium, and the OD values ​​of the two strains were measured in the same manner as in Example 1. 600 After normalization, the culture was incubated at 37°C for 18 h, and the supernatant was collected by centrifugation and passed through a membrane (with a pore size of 0.22 μm). High performance liquid chromatography (HPLC) was used to detect the lactic acid production.

[0050] The lactic acid production of Lactobacillus acidophilus CCNH655 is 18.67 g / L, while that of LGG is 15.1 g / L. Under the same conditions, the lactic acid production performance of strain CCNH655 activated is better than that of LGG, indicating that strain Lactobacillus acidophilus CCNH655 can metabolize and decompose more sugars during the culture process, thereby reducing the sugar content in the environment.

[0051] Example 4 Take the cultured bacteria, wash twice with saline, and resuspend in 0.1M PBS to make the OD of the bacterial suspension 600 =1, pipette 100 μL of bacterial suspension into the ELISA plate and detect the initial OD 600 The bacterial solution was allowed to stand for 4 hours, and 100 μL of the upper bacterial solution was taken to the ELISA plate to detect the OD value after standing. 600 value, recorded as A1; take 900 μL of the above bacterial suspension, add 180 μL of dodecane, vortex until mixed evenly, let stand for 20 minutes to separate the layers, take the lower aqueous phase and measure its OD 600 The value is recorded as A2, and the self-aggregation rate or hydrophobicity is calculated according to the following formula: Self-aggregation rate or hydrophobicity rate = (A0-A1 or A2) / A0×100%.

[0052] The self-aggregation rates of Lactobacillus acidophilus CCNH655 and LGG were 59.41% and 54.55%, and the hydrophobicity rates were 50.21% and 43.85%, respectively. The two data combined show that compared with LGG, Lactobacillus acidophilus CCNH655 has better adhesion, is easier to bind to intestinal epithelial cells and colonize in the intestine, thereby better inhibiting the colonization of pathogens in the gastrointestinal tract.

[0053] Example 5 Weigh 44 g of Columbia blood agar medium powder (purchased from Qingdao Haibo Biological, catalog number HB9295), add 1000 mL of water, sterilize at 121°C for 15 min, and then add 10 vol% sterile defibrinated sheep blood to obtain a Columbia blood agar plate.

[0054] 10 μL of the culture solution was inoculated on the Columbia blood agar plate. In addition, Escherichia coli ATCC 25922 and Staphylococcus aureus CMCC (B) 26001 with the same number of viable cells were used as control strains.

[0055] The results showed that Lactobacillus acidophilus CCNH655 and Escherichia coli ATCC 25922 were not hemolytic, while Staphylococcus aureus CMCC 26001 was completely hemolytic, with β-hemolytic zones around white colonies, indicating that Lactobacillus acidophilus CCNH655 provided by the present invention would not cause hemolysis, and thus has the possibility of in vivo application.

[0056] Example 6 The culture solution was centrifuged and the supernatant was taken. 25 μL of nitrophenyl-β-D-glucoside (PNPG, 20 mmol / L) and 25 μL of supernatant were added to 25 μL of PBS buffer (0.1 mol / L, pH=6.8) as the sample group. After 10 min at 37°C, 50 μL of α-glucosidase (0.2 U / mL, purchased from Sigma, catalog number G5003) was added, and the reaction was carried out at 37°C for 15 min. Finally, 100 μL of 0.1 mol / L sodium carbonate solution was added to terminate the reaction. In addition, a sample control group (no α-glucosidase), a blank group (no supernatant), a blank control group (no α-glucosidase and supernatant) and a positive control group (acarbose, an α-glucosidase inhibitor, 1 μg / μL) were set up respectively.

[0057] The absorbance of the samples detected at 405 nm is A 样品 , A 样品对照 , A 空白 and A 空白对照 The inhibition rate was calculated by the following formula: Inhibition rate = (1-(A 样品 +A 样品对照 )) / (A 空白 -A 空白对照 )×100%, unit is %.

[0058] The α-glucosidase inhibition rates of Lactobacillus acidophilus CCNH655 and LGG were 67.19% and 26.46%, respectively, and the α-glucosidase inhibition rate of acarbose was 78.42%. The results showed that the strain Lactobacillus acidophilus CCNH655 can inhibit the activity of α-glucosidase, and thus can be used to lower blood sugar by inhibiting the activity of α-glucosidase.

[0059] Example 7 Take 1 mL of the culture solution, centrifuge to remove the supernatant, wash the bacteria twice with saline, and resuspend them in 360 μL of PBS solution to obtain a bacterial suspension.

[0060] Catalytic system: Sodium glycocholate and sodium taurocholate are used as substrates to simulate the decomposition of cholesterol. After decomposition, glycine and taurine are generated respectively. 20 μL of sodium glycocholate solution and sodium taurocholate solution (both concentrations are 200 mM, the solvent is water) are added to two 180 μL bacterial suspensions respectively, react at 37°C for 3 hours, and centrifuge to collect the degradation supernatant.

[0061] Prepare glycine and taurine solutions with concentrations of 0.1, 0.2, 0.5, 0.8, 1.0, 2.0, 4.0 and 5.0 mM, respectively, detect the absorbance of the two at different concentrations, and establish standard curves for glycine and taurine. Prepare a liquid containing 20 μL of degradation supernatant according to the requirements of the kit, boil the liquid in a water bath for 15 minutes, draw 200 μL, and use the amino acid detection kit AA-W96-N (1620) to detect the absorbance at 570 nm to determine the concentration of sodium glycocholate and sodium taurocholate. Bile salt hydrolase activity (U / mL bacterial suspension): The amount of enzyme that releases amino acids from the substrate within 1 minute per 1 mL bacterial suspension.

[0062] In the experiments of Lactobacillus acidophilus CCNH655 group and LGG group, the bile salt hydrolase activities with sodium glycocholate as substrate were 3.89U / mL and 2.27U / mL, respectively, and the bile salt hydrolase activities with sodium taurocholate as substrate were 6.91U / mL and 4.98U / mL, respectively.

[0063] Example 8 Ordinary feed Chowdiet, fat content 10wt%; High-fat diet HFD, fat content 40wt%; Acarbose was purchased from Bayer Healthcare Co., Ltd.; Streptozotocin (STZ) was purchased from Merck, Darmstadt, Germany.

[0064] Fifty SPF male C57BL / 6J mice, 6 weeks old, were selected and housed in an SPF laboratory animal center at room temperature of 23-25°C, appropriate humidity, and a 12-h day-night cycle. After one week of adaptive feeding, the mice were randomly divided into 5 groups, 10 mice in each group, namely: (1) Normal control group: fed with normal feed, without STZ injection, intraperitoneal injection of 0.1 mmol / L citrate buffer (pH = 4.4) on the first day of the fifth week, and intragastric administration of 300 uL PBS every day from the sixth to the thirteenth week; (2) Model group: After being fed a high-fat diet for 5 weeks, STZ was injected on the first day, and 300uL of PBS was gavaged daily from the 6th to the 13th week; (3) Positive drug group: After 5 weeks of high-fat diet, STZ was injected on the first day, and acarbose was used for treatment from week 6 to week 13 at a dosage of 100 mg / kg body weight; (4) LGG group: After being fed a high-fat diet for 5 weeks, STZ was injected on the first day, and strain LGG was used for intervention from 6 to 13 weeks; (5) CCNH655 group: After being fed a high-fat diet for 5 weeks, STZ was injected on the first day, and the CCNH655 strain was used for intervention from 6 to 13 weeks.

[0065] The intervention method of strains was oral administration, and probiotics were given by oral administration every day (1×10 9 CFU, dissolved in 300 μL PBS buffer); STZ was injected intraperitoneally at a dose of 50 mg / kg body weight. Blood glucose was measured for all mice 7 days after STZ injection. If the fasting blood glucose of the mice was ≥7 mmol / L or the postprandial blood glucose was ≥11.1 mmol / L, the diabetes model was considered to be successfully established.

[0066] From week 6 to week 13, mice in all groups received normal diet and were gavaged with corresponding drugs every day for a total of 8 weeks. Food intake, water intake, and body weight were monitored weekly during the experiment.

[0067] The detection methods of blood sugar and blood lipid related indicators are as follows: (1) Fasting blood glucose: After the mice were fasted for 16 h, blood was collected from the tail tip and blood glucose was measured using a blood glucose meter. (2) Insulin: Detect plasma insulin levels using an ELISA test kit; (3) Glucose tolerance test (GTT): The oral glucose tolerance of all mice was measured one week after modeling and one week before sacrifice. The specific method is: after fasting for 16 hours, the mice were intraperitoneally injected with glucose solution (1.0 g / kg body weight), and blood was collected from the tail tip at 0, 30, 60, 90, and 120 minutes after the injection of glucose. Blood glucose was measured with a blood glucose meter, and the area under the curve (AUC) of the GTT time-blood glucose curve was calculated. The AUC value directly reflects the glucose tolerance of the mice; (4) Blood lipid related indicators: The corresponding detection kits of triglycerides, total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were used to measure the levels of triglycerides, total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol in mouse serum; (5) Other indicators: Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and glucagon-like peptide-1 (GLP-1) in mouse serum were detected by ELISA according to the instructions of the kit, and their contents were determined by an enzyme-labeled instrument.

[0068] The average body weight, fasting blood glucose, glucose tolerance (expressed as AUC index) and GLP-1 test results of mice after 8 weeks of intervention are shown in Table 1.

[0069] Table 1

[0070] Note: In different groups of data under the same parameter, the same letters indicate no significant difference between the groups ( P>0.05), different letters indicate significant differences among groups ( P <0.05).

[0071] The body weight, fasting blood glucose and AUC index of the model group were significantly higher than those of the control group. Compared with the model group, the CCNH655 group could significantly reduce the body weight, fasting blood glucose and AUC levels of mice.

[0072] The GLP-1 level in the model group was significantly lower than that in the control group, and the GLP-1 level in the CCNH655 group was significantly increased, indicating that CCNH655 can promote the synthesis and secretion of insulin and regulate glucose metabolism homeostasis.

[0073] The levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in mice after 8 weeks of intervention are shown in Table 2 (unit: mmol / L).

[0074] Table 2

[0075] Note: In different groups of data under the same parameter, the same letters indicate no significant difference between the groups ( P >0.05), different letters indicate significant differences among groups ( P <0.05).

[0076] Compared with the normal control group, the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the serum of the model group were significantly increased, while the level of high-density lipoprotein cholesterol was significantly decreased. Compared with the model group, the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the CCNH655 group were significantly decreased, while the level of high-density lipoprotein cholesterol was significantly increased, indicating that CCNH655 can effectively inhibit lipid metabolism disorders in diabetic mice.

[0077] The levels of inflammatory factors in the serum of mice after 8 weeks of intervention are shown in Table 3.

[0078] Table 3

[0079] Note: In different groups of data under the same parameter, the same letters indicate no significant difference between the groups ( P >0.05), different letters indicate significant differences among groups ( P <0.05).

[0080] Inflammatory factors play a very important role in the pathogenesis of type 2 diabetes. Among them, tumor necrosis factor-α (TNF-α) can cause insulin resistance, and interleukin-6 (IL-6) can toxicize pancreatic islet cells, thereby accelerating the progression of diabetes. Compared with the normal control group, the serum levels of interleukin-6 and tumor necrosis factor-α in the model group mice were significantly increased, while those in the CCNH655 group were significantly lower than those in the model group.

[0081] Example 9 After centrifuging the cultured bacterial solution, take the supernatant. Add 25μL of PBS buffer (0.1mol / L, pH=6.8), 50μL of α-amylase solution, 50μL of 1wt% soluble starch solution, and 25μL of fermentation broth centrifugation supernatant as the sample group. After reacting at 37°C for 30 minutes, add 1μL of iodine solution and detect the absorbance at 660nm. Set up a sample control group (without adding α-amylase), a blank group (without adding supernatant) and a blank control group (without adding α-amylase and supernatant). The sample absorbance detected at 660nm is A sample, A sample control, A blank and A blank control, respectively. The inhibition rate is calculated by the following formula. In addition, acarbose (1μg / μL) is set as the positive control group. Inhibition rate = (1-(A 样品 +A 样品对照 )) / (A 空白 -A 空白对照 )×100%.

[0082] The α-amylase inhibition rate of the positive control group was 73.47%, and the α-amylase inhibition rates of Lactobacillus acidophilus CCNH655 and LGG were 86.75% and 81.45%, respectively, indicating that the strain Lactobacillus acidophilus CCNH655 can inhibit the activity of α-amylase and thus can be used to lower blood sugar by inhibiting the activity of α-amylase.

[0083] Example 10 Cholesterol solution: 0.1g cholesterol, 0.2g ox bile salt, 0.11g sucrose ester, 1mL Tween 80, 5mL anhydrous ethanol, add to a 15mL centrifuge tube. Ultrasonic treatment, after 95±5℃ hot water bath, 80W ultrasonic for 20min, vortex mix every 4-8min until the cholesterol is completely dissolved, and sterilize by membrane while hot (membrane pore size is 0.22μm).

[0084] Add 360 μL of the above cholesterol solution to 30 mL of MRS liquid culture medium while hot, stir magnetically at 800 rpm for 15 min. Inoculate MRS liquid culture medium with Lactobacillus acidophilus CCNH655 and LGG glycerol tubes, normalize OD, transfer 2 vol% to the above MRS liquid culture medium containing cholesterol, and culture at 37°C and 180 rpm for 24 h. The blank culture medium is used as a blank control. The cholesterol content in the supernatant of the fermentation broth was detected at 500 nm using the cholesterol detection kit TC-W48-N (1031). The cholesterol degradation rate was calculated according to the following formula: Degradation rate = (A 空白对照 -A 样品 ) / A 空白对照 ×100%.

[0085] The calculated cholesterol clearance rates were 79.39% and 34.06%, respectively, indicating that the cholesterol degradation ability of Lactobacillus acidophilus CCNH655 was stronger than that of LGG.

[0086] Embodiment 11 Escherichia coli (CICC 10421 and ATCC 25922), Staphylococcus aureus (CMCC(B)26001 and CMCC(B)26003), Salmonella (ATCC 14028) and CVCC 3378) were inoculated into LB liquid medium, and Candida albicans (CICC 1965 and ATCC 10231) was inoculated into YPD solid medium. They were cultured aerobically at 37°C and activated for 18 h. They were then transferred to the corresponding fresh medium and cultured until the number of viable cells reached 10 5 -10 7 CFU / mL, dilute to 10 5 CFU / mL was used as the indicator bacterial solution. The culture solution was centrifuged and the supernatant was taken and mixed with 10 5 The bacterial suspension of pathogenic bacteria with a volume ratio of 1:1 was mixed, and the supernatant was replaced with an equal volume of sterile MRS liquid culture medium as a negative control group, and the fermentation supernatant was replaced with 10mL of kanamycin (content 100μg / mL) as a positive control group. Measure the OD of the experimental group and the negative control group 600 The values ​​are A and A respectively. 0， The inhibition rate was calculated according to the following formula and the results are shown in Table 4.

[0087] Inhibition rate (%) = (A0-A) / A0×100%.

[0088] Table 4

[0089] The results showed that Lactobacillus acidophilus CCNH655 had a certain inhibitory ability against Escherichia coli, Staphylococcus aureus and Salmonella.

[0090] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A strain of Lactobacillus acidophilus Lactobacillus acidophilus , characterized in that, The deposit number of the Lactobacillus acidophilus is CGMCC No.33155.

2. A bacterial agent, characterized in that The bacterial agent contains the Lactobacillus acidophilus according to claim 1.

3. The bacterial agent according to claim 2, wherein The bacterial agent contains auxiliary materials, wherein the auxiliary materials are selected from glycerol and / or corn oil.

4. The bacterial agent according to claim 2 or 3, wherein The number of viable Lactobacillus acidophilus in the bacterial agent is not less than 10 4 CFU / mL.

5. Use of the Lactobacillus acidophilus according to claim 1 in the preparation of a medicament for decomposing cholesterol.

6. Use of the Lactobacillus acidophilus according to claim 1 in the preparation of a medicament for inhibiting the activity of α-glucosidase.

7. Use of the Lactobacillus acidophilus according to claim 1 in the preparation of a medicament for preventing and / or treating hyperlipidemia.

8. Use of the Lactobacillus acidophilus according to claim 1 in the preparation of a medicament for preventing and / or treating hyperglycemia.

9. Use of the Lactobacillus acidophilus according to claim 1 in the preparation of a medicament for reducing inflammatory factors.

10. The use according to any one of claims 5 to 9, wherein: The drug further comprises pharmaceutically acceptable excipients, including corn oil and / or glycerol.

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