Lactobacillus acidophilus and its application in the preparation of drugs for preventing and / or treating hyperlipidemia and hyperglycemia

Abstract

The present invention relates to the field of microorganisms, and discloses a Lactobacillus acidophilus and its application in the preparation of drugs for preventing and / or treating hyperlipidemia and hyperglycemia. The preservation number of the Lactobacillus acidophilus is CGMCC No. 33155. This strain can produce lactic acid, be acid-tolerant and bile-salt tolerant, and has strong auto-aggregation ability and hydrophobic ability. The strain provided by the present invention can promote the decomposition of fat and sugar by degrading cholesterol and inhibiting the activities of α-glucosidase and α-amylase, and can reduce the blood glucose, blood lipid and inflammatory factor expression levels of hyperlipidemic and hyperglycemic mice, providing a new idea for the preparation of drugs for preventing and / or treating hyperglycemia and hyperlipidemia.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, and particularly to a strain of Lactobacillus acidophilus and its application in the preparation of drugs for preventing and / or treating hyperlipidemia and hyperglycemia. Background Art

[0002] With the improvement of people's living standards, high-sugar and high-fat diets have become common. However, due to reasons such as irregular and unrestrained diets, cardiovascular diseases such as hyperglycemia, hyperlipidemia, and hypertension have followed. It is reported that the mortality rate of cardiovascular diseases ranks first, higher than that of tumors and other diseases. Now, the group of people with "three highs" diseases has gradually become younger, and the prevention and treatment of "three highs" diseases are imminent.

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

[0004] Therefore, there is an urgent need to select and breed strains with excellent fermentation characteristics, high viable counts, certain probiotic functions, and that can stably survive in the human body and can be used to prepare drugs for preventing and / or treating 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 provide a strain of Lactobacillus acidophilus and its application in the preparation of drugs for preventing and / or treating hyperlipidemia and hyperglycemia.

[0006] The first aspect of the present invention provides a strain of Lactobacillus acidophilus Lactobacillus acidophilus , and the preservation number of the Lactobacillus acidophilus is CGMCC No. 33155.

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

[0008] The third aspect of the present invention provides the application of the Lactobacillus acidophilus as described above in the preparation of drugs for decomposing cholesterol.

[0009] The fourth aspect of the present invention provides the application of the Lactobacillus acidophilus as described above in the preparation of drugs for inhibiting the activity of α-glucosidase.

[0010] The fifth aspect of the present invention provides the application of the Lactobacillus acidophilus as described above in the preparation of drugs for preventing and / or treating hyperlipidemia.

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

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

[0013] Through the above technical solutions, the beneficial effects of the present invention at least include:

[0014] The Lactobacillus acidophilus of the present invention has the ability to tolerate gastric acid and bile salts, can decompose saccharide substances into high levels of lactic acid, and has high inhibition rates of α-glucosidase and α-amylase and high cholesterol clearance rate. It can reduce blood glucose, blood lipids, and inflammatory factors in hyperlipidemic and hyperglycemic mice, and has the prospect of being used to prepare medicaments for preventing and / or treating hyperglycemia and hyperlipidemia.

[0015] Biological deposit

[0016] The strain provided by the present invention is taxonomically named Lactobacillus acidophilus Lactobacillus acidophilus , and was deposited at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms (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.

[0017] The strain provided by the present invention is taxonomically named Lactobacillus plantarum Lactobacillus plantarum , and was deposited at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms (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. Description of the drawings

[0018] 图1 is a morphological diagram of the colony of Lactobacillus acidophilus provided by the present invention;

[0019] 图2 is a Gram staining microscopic examination diagram of the colony of Lactobacillus acidophilus provided by the present invention. Detailed implementation manners

[0020] In the ranges disclosed herein, the endpoints and any values are not limited to the exact ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0021] The inventor of the present invention accidentally isolated a strain of Lactobacillus acidophilus, as 图1 and 图2 As shown, the colonies of this strain are punctate, white and opaque, with a plump, smooth and moist morphology, and the bacterial lawn grows along the streak. It is determined to be Lactobacillus acidophilus and named Lactobacillus acidophilus CCNH655.

[0022] Based on the above findings, in the first aspect of the present invention, a strain of Lactobacillus acidophilus is provided Lactobacillus acidophilus , and the preservation number of the Lactobacillus acidophilus is CGMCC No. 33155.

[0023] In the second aspect of the present invention, a bacterial agent is provided, and the bacterial agent contains the Lactobacillus acidophilus as described above.

[0024] The present invention has no particular limitation on the type of the bacterial agent, and it can be any existing bacterial agent type 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.

[0025] In the present invention, the bacterial agent may further contain excipients, and the excipients may be protective agents (such as lyophilization protectants, such as skim milk powder, maltodextrin, trehalose, dextran, corn oil and glycerol, etc.) and / or buffering agents (such as the buffer solution remaining in the process of preparing the solid bacterial agent, etc.). Preferably, the excipients may be selected from glycerol and / or corn oil.

[0026] In the present invention, the viable count of Lactobacillus acidophilus in the bacterial agent may be not less than 10 4 CFU / mL.

[0027] In the third aspect of the present invention, the application of the Lactobacillus acidophilus as described above in the preparation of a drug for decomposing cholesterol is provided.

[0028] Preferably, the cholesterol is glycocholic acid and / or taurocholic acid.

[0029] In the fourth aspect of the present invention, the application of the Lactobacillus acidophilus as described above in the preparation of a drug for inhibiting the activity of α-glucosidase is provided.

[0030] In the fifth aspect of the present invention, the application of the Lactobacillus acidophilus as described above in the preparation of a drug for preventing and / or treating hyperlipidemia is provided.

[0031] In the sixth aspect of the present invention, the application of the Lactobacillus acidophilus as described above in the preparation of a drug for preventing and / or treating hyperglycemia is provided.

[0032] According to the present invention, the hyperglycemia is caused by type II diabetes.

[0033] According to an embodiment of the present invention, the strain provided by the present invention can improve the physiological indexes of mice already suffering from diabetes. Therefore, the strain provided by the present invention can be used in the application of preparing a drug for treating hyperglycemia caused by diabetes.

[0034] The seventh aspect of the present invention provides the application of Lactobacillus acidophilus as described above in the preparation of a drug for reducing inflammatory factors.

[0035] Preferably, the inflammatory factors include tumor necrosis factor-α (TNF-α) and / or interleukin-6 (IL-6).

[0036] In the present invention, the drug may contain pharmaceutically acceptable excipients, including corn oil and / or glycerol.

[0037] The present invention also provides the application of Lactobacillus plantarum as described above in the preparation of a drug for inhibiting the reproduction of pathogenic bacteria.

[0038] According to the present invention, the pathogenic bacteria may include Escherichia coli ( Escherichia coli ), Staphylococcus aureus ( Staphylococcus aureus ), Salmonella ( Salmonella ), and Candida albicans ( Candida albicans ) at least one of them.

[0039] According to some embodiments of the present invention, the Salmonella preferably includes Salmonella typhimurium ( Salmonella typhimurium ) and Salmonella enteritidis ( Salmonella enteritidis ).

[0040] The present invention also provides a compound bacterium agent, which includes Lactobacillus acidophilus with the preservation number of CGMCC No. 33155 provided by the present invention and Lactobacillus plantarum with the preservation number of CCGMCC No. 33152 Lactobacillus plantarum 。

[0041] As shown in the records of the application document with the application number of 202510222543.6, Lactobacillus plantarum with the preservation number of CCGMCC No. 33152 has the effects of antibacterial, inhibiting the reproduction of pathogenic bacteria, and reducing the expression of pro-inflammatory factors, and also has biological safety and antibiotic sensitivity. Therefore, when using Lactobacillus plantarum with the preservation number of CCGMCC No. 33152 to prepare a drug for preventing and / or treating vaginitis, a drug prepared from Lactobacillus acidophilus with the preservation number of CGMCC No. 33155 provided by the present application can be used in combination to further enhance the antibacterial effect and anti-inflammatory effect. That is, Lactobacillus plantarum with the preservation number of CCGMCC No. 33152 and Lactobacillus acidophilus provided by the present application can be compounded into a compound bacterium agent to prepare a drug for preventing and / or treating vaginitis.

[0042] In the above-mentioned composite bacterial agent provided by the present invention, the two strains can be independently packaged or mixedly packaged. However, in order to maintain the activity of the strains to the greatest extent and exert the effect, it is preferably independently packaged for each.

[0043] When a patient uses a drug containing the above-mentioned composite bacterial agent provided by the present invention, a drug containing the two strains can be used simultaneously or successively. However, in order to exert the effect to the greatest extent, it is preferably used successively, but the order of successive use is not limited.

[0044] The present invention will be described in detail below through examples.

[0045] In the following examples, without special instructions, the chemical / biological reagents used are all obtained by purchasing from regular chemical / biological reagent suppliers.

[0046] Lactobacillus rhamnosus Lactobacillus rhamnosus ) LGG, numbered ATCC 53103, was purchased from the American Type Culture Collection;

[0047] The MRS liquid medium was purchased from Qingdao Haibo Biotechnology, with the product number HB0384-1;

[0048] The PBS buffer solution was purchased from Solarbio Science & Technology Co., Ltd., with the product number P1003;

[0049] Escherichia coli ( Escherichia coli ), numbered (1) ATCC 25922, was purchased from the American Type Culture Collection, and (2) CICC 10421, was purchased from the China Center for Industrial Culture Collection;

[0050] Staphylococcus aureus ( Staphylococcus aureus ), numbered CMCC(B)26001 and CMCC(B)26003, were both purchased from the China National Center for Medical Culture Collections;

[0051] Salmonella, (1) Salmonella typhimurium ( Salmonella typhimurium ), numbered ATCC 14028, was purchased from the American Type Culture Collection; (2) Salmonella enteritidis ( Salmonella enteritidis ), numbered CVCC 3378, was purchased from the National Veterinary Culture Collection Center;

[0052] Candida albicans ( Candida albicans ), numbered (1) CICC 1965, was purchased from the China Center for Industrial Culture Collection, and (2) ATCC 10231, was purchased from the American Type Culture Collection.

[0053] Example 1

[0054] Inoculate the glycerol tubes of Lactobacillus acidophilus CCNH655 and LGG into fresh MRS liquid medium at 2 vol% respectively, and culture them overnight at 37 °C for 18 h. Measure the OD of the two strains 600 value. Add MRS liquid medium to the medium to make the OD of the two strains 600 equal, obtaining an activated bacterial solution. Inoculate it into fresh MRS liquid medium at an inoculation amount of 2 vol%, and culture it at 37 °C and 200 rpm for 18 h to obtain an expanded culture solution of Lactobacillus acidophilus CCNH655 and an expanded culture solution of LGG.

[0055] Measure the OD of the expanded culture solution of Lactobacillus acidophilus CCNH655 and the expanded culture solution of LGG 600 which are 8.71 and 8.13 respectively. The growth performance of Lactobacillus acidophilus CCNH655 is better than that of LGG.

[0056] Example 2

[0057] Take the expanded culture solutions of Lactobacillus acidophilus CCNH655 and LGG, and refer to T / CNHFA435-2024 "Detection Method for Gastric Juice Tolerance of Probiotic Agents" to evaluate the gastric acid tolerance and bile salt tolerance of the strains.

[0058] In the gastric acid tolerance experiment, the pH is 3 and the incubation time is 2 h; in the bile salt tolerance experiment, the bile salt content is 0.2 wt% and the incubation time is 4 h.

[0059] The survival rates of Lactobacillus acidophilus CCNH655 and LGG in the gastric acid tolerance experiment are 95.48% and 93.33% respectively, and the survival rates in the bile salt tolerance experiment are 75.21% and 53.13% respectively. It can be found that the gastric acid tolerance and bile salt tolerance of Lactobacillus acidophilus CCNH655 and LGG are both better than those of LGG.

[0060] Example 3

[0061] Inoculate 600 μL of the activated bacterial solution into 30 mL of fresh MRS liquid medium. After normalizing the OD of the two strains in the manner of Example 1 600 and culturing it statically at 37 °C for 18 h, centrifuge to obtain the supernatant, and perform membrane filtration treatment (membrane pore size is 0.22 μm). Use high performance liquid chromatography HPLC to detect the lactic acid production.

[0062] The lactic acid production of Lactobacillus acidophilus CCNH655 is 18.67 g / L, while the lactic acid production of LGG is 15.1 g / L. Under the same activation conditions, the lactic acid production performance of strain CCNH655 is better than that of LGG, indicating that Lactobacillus acidophilus CCNH655 can metabolize and decompose more sugars during the culture process, thereby reducing the sugar content in the environment.

[0063] Example 4

[0064] Take the expanded culture broth, wash it twice with physiological saline, and resuspend it with 0.1M PBS to make the OD of the bacterial suspension 600 =1. Pipette 100 μL of the bacterial suspension into an ELISA plate and measure the initial OD 600 value, denoted as A0; let the bacterial solution stand for 4 h, take 100 μL of the upper-layer bacterial solution and transfer it to an ELISA plate, and measure its OD after standing 600 value, denoted as A1; pipette 900 μL of the above-mentioned bacterial suspension, add 180 μL of dodecane, vortex and oscillate until evenly mixed, let it stand for 20 min to separate layers, and measure the OD of the lower-layer aqueous phase 600 value, denoted as A2, and calculate the self-aggregation rate or hydrophobicity rate according to the following formula:

[0065] Self-aggregation rate or hydrophobicity rate = (A0 - A1 or A2) / A0 × 100%.

[0066] The self-aggregation rates of Lactobacillus acidophilus CCNH655 and LGG are 59.41% and 54.55% respectively, and the hydrophobicity rates are 50.21% and 43.85% respectively. The two sets of data comprehensively show that compared with LGG, Lactobacillus acidophilus CCNH655 has a better adhesion effect, is more likely to bind to intestinal epithelial cells and colonize in the intestine, thereby better inhibiting the colonization of pathogenic bacteria in the gastrointestinal tract.

[0067] Example 5

[0068] Weigh 44 g of Columbia blood agar medium powder (purchased from Qingdao Hope Bio-Technology Co., Ltd., product number HB9295), add 1000 mL of water, sterilize at 121 °C for 15 min, and then add 10 vol% of sterile defibrinated sheep blood to obtain a Columbia blood agar plate.

[0069] Inoculate 10 μL of the expanded culture broth on the above-mentioned Columbia blood agar plate. In addition, Escherichia coli ATCC 25922 and Staphylococcus aureus CMCC(B)26001 with the same viable bacteria count are used as control strains.

[0070] The results show that Lactobacillus acidophilus CCNH655 and Escherichia coli ATCC 25922 do not cause hemolysis, while Staphylococcus aureus CMCC 26001 causes complete hemolysis, with a β-hemolysis zone around the white colonies. This indicates that Lactobacillus acidophilus CCNH655 provided by the present invention does not cause hemolysis, thus having the possibility of in vivo application.

[0071] Example 6

[0072] Centrifuge the expanded bacterial solution and take the supernatant. Add 25 μL of nitro phenyl-β-D-glucoside (PNPG, 20 mmol / L) and 25 μL of the supernatant to 25 μL of PBS buffer (0.1 mol / L, pH = 6.8) as the sample group. After reacting at 37 °C for 10 min, add 50 μL of α-glucosidase (0.2 U / mL, purchased from Sigma, catalog number G5003), react at 37 °C for 15 min, and finally add 100 μL of 0.1 mol / L sodium carbonate solution to terminate the reaction. In addition, set up a sample control group (without adding α-glucosidase), a blank group (without adding the supernatant), a blank control group (without adding α-glucosidase and the supernatant), and a positive control group (acarbose, an α-glucosidase inhibitor, 1 μg / μL).

[0073] The absorbances of the samples were detected at 405 nm as A 样品 , A 样品对照 , A 空白 and A 空白对照 , respectively, and 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%, with the unit of %.

[0074] 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%. These results indicate that the strain Lactobacillus acidophilus CCNH655 can reduce blood glucose by inhibiting the activity of α-glucosidase.

[0075] Example 7

[0076] Take 1 mL of the expanded bacterial solution, centrifuge to remove the supernatant, wash the bacterial cells twice with physiological saline, and resuspend with 360 μL of PBS solution to obtain a bacterial suspension.

[0077] Catalytic system: Sodium glycocholate and sodium taurocholate are used as substrates for simulating cholesterol decomposition, and glycine and taurine are generated respectively after their decomposition. Add 20 μL of sodium glycocholate solution and sodium taurocholate solution (both with a concentration of 200 mM and the solvent is water) to two portions of 180 μL of the bacterial suspension respectively, react at 37 °C for 3 h, and centrifuge to collect the degradation supernatant.

[0078] 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 the standard curves of glycine and taurine. Prepare a liquid containing 20 μL of the degradation supernatant according to the requirements of the kit, boil the liquid in a water bath for 15 min, aspirate 200 μL, and use the amino acid detection kit AA-W96-N (1620) to detect the absorbance at 570 nm to determine the concentrations of sodium glycochenodeoxycholate and sodium taurocholate. Bile salt hydrolase activity (U / mL bacterial suspension): the amount of enzyme that releases amino acids from the substrate per 1 mL of bacterial suspension in 1 min.

[0079] In the experiments of Lactobacillus acidophilus CCNH655 group and LGG group, the bile salt hydrolase activities using sodium glycochenodeoxycholate as the substrate were 3.89 U / mL and 2.27 U / mL respectively, and the bile salt hydrolase activities using sodium taurocholate as the substrate were 6.91 U / mL and 4.98 U / mL respectively.

[0080] Example 8

[0081] Ordinary feed Chowdiet, with a fat content of 10 wt%;

[0082] High-fat feed HFD, with a fat content of 40 wt%;

[0083] Acarbose, purchased from Bayer Healthcare Co., Ltd.;

[0084] Streptozotocin STZ, purchased from Merck KGaA, Darmstadt, Germany.

[0085] Select 50 SPF-grade male C57BL / 6J mice, 6 weeks old, and raise them in an SPF-grade experimental animal center at a room temperature of 23 - 25 °C with appropriate humidity and a 12 h day-night cycle. After 1 week of adaptive feeding of the mice, randomly divide the mice into 5 groups, with 10 mice in each group, namely:

[0086] (1) Normal control group: Fed with ordinary feed, not injected with STZ, intraperitoneally injected with 0.1 mmol / L citrate buffer (pH = 4.4) on the 1st day of the 5th week, and gavaged with 300 μL of PBS every day from the 6th to the 13th week;

[0087] (2) Model group: Fed with high-fat feed until the 5th week, injected with STZ on the 1st day, and gavaged with 300 μL of PBS every day from the 6th to the 13th week;

[0088] (3) Positive drug group: Fed with high-fat feed for 5 weeks, injected with STZ on the 1st day, and treated with acarbose from the 6th to the 13th week at a dosage of 100 mg / kg body weight;

[0089] (4)LGG group: After feeding with high-fat diet for 5 weeks, STZ was injected on the 1st day, and the mice were intervened with strain LGG from the 6th to the 13th week;

[0090] (5)CCNH655 group: After feeding with high-fat diet for 5 weeks, STZ was injected on the 1st day, and the mice were intervened with strain CCNH655 from the 6th to the 13th week.

[0091] The intervention method of the strain was intragastric administration, and probiotics were given intragastrically every day (1×10 9 CFU, dissolved in 300 μL of PBS buffer); the injection method of STZ was intraperitoneal injection, and the injection volume was 50 mg / kg body weight. Seven days after the injection of STZ, the blood glucose of all mice was measured. If the fasting blood glucose of the mice was ≥7 mmol / L or the postprandial blood glucose was ≥11.1 mmol / L, it was considered that the diabetes model was successfully established.

[0092] From the 6th to the 13th week, all groups of mice received normal diet, and at the same time were given corresponding drugs intragastrically every day for a total of 8 weeks. During the experiment, the food intake, water intake, and body weight were monitored weekly.

[0093] The detection methods of blood glucose and blood lipid-related indicators are as follows:

[0094] (1)Fasting blood glucose: After the mice were fasted for 16 h, blood was taken from the tip of the tail, and the blood glucose was measured with a blood glucose meter;

[0095] (2)Insulin: The insulin level in plasma was detected with an ELISA detection kit;

[0096] (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 was as follows: After the mice were fasted for 16 h, a glucose solution (1.0 g / kg body weight) was intraperitoneally injected, and blood was taken from the tip of the tail at 0, 30, 60, 90, and 120 minutes after the injection of glucose, and the 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 size of the AUC value directly reflected the glucose tolerance of the mice;

[0097] (4)Blood lipid-related indicators: The contents of triglyceride, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol in the serum of mice were measured with corresponding detection kits for triglyceride, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol;

[0098] (5)Other indicators: Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and glucagon-like peptide-1 (GLP-1) in the serum of mice were detected by ELISA according to the kit instructions, and their contents were measured by an enzyme-linked immunosorbent assay reader.

[0099] After 8 weeks of intervention, the average body weight, fasting blood glucose, glucose tolerance (represented by the AUC index), and GLP-1 test results of the mice are shown in Table 1.

[0100] Table 1

[0101]

[0102] Note: Among the different groups of data under the same parameters, data with the same letter indicate no significant difference between groups ( P > 0.05), and different letters indicate a significant difference between groups ( P < 0.05).

[0103] 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 the mice.

[0104] The GLP-1 of the model group was significantly lower than that of the control group, and the GLP-1 level of the CCNH655 group was significantly increased, indicating that CCNH655 can promote the synthesis and secretion of insulin and regulate glucose metabolism homeostasis.

[0105] After 8 weeks of intervention, the levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in the mice are shown in Table 2 (unit: mmol / L).

[0106] Table 2

[0107]

[0108] Note: Among the different groups of data under the same parameters, data with the same letter indicate no significant difference between groups ( P > 0.05), and different letters indicate a significant difference between groups ( P < 0.05).

[0109] Compared with the normal control group, the contents of total cholesterol, triglyceride, and low-density lipoprotein cholesterol in the serum of the model group were significantly increased, while the content of high-density lipoprotein cholesterol was significantly decreased. Compared with the model group, the contents of total cholesterol, triglyceride, and low-density lipoprotein cholesterol in the CCNH655 group were significantly decreased, while the content of high-density lipoprotein cholesterol was significantly increased, indicating that CCNH655 can effectively inhibit lipid metabolism disorders in diabetic mice.

[0110] After 8 weeks of intervention, the inflammatory factor conditions in the serum of the mice are shown in Table 3.

[0111] Table 3

[0112]

[0113] Note: In different groups of data under the same parameters, the same letter indicates no significant difference between groups ( P > 0.05), and different letters indicate a significant difference between groups ( P < 0.05).

[0114] 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, while interleukin-6 (IL-6) can be toxic to pancreatic islet cells, thereby accelerating the progression of diabetes. Compared with the normal control group, the levels of interleukin-6 and tumor necrosis factor-α in the serum of the model group mice were significantly increased, while those in the CCNH655 group were significantly lower than those in the model group.

[0115] Example 9

[0116] After centrifuging the expanded culture broth, take the supernatant. Add 25 μL of PBS buffer (0.1 mol / L, pH = 6.8), 50 μL of α-amylase solution, 50 μL of 1 wt% soluble starch solution, and 25 μL of the centrifuged supernatant of the fermentation broth as the sample group. After reacting at 37 °C for 30 min, add 1 μL of iodine solution and measure the absorbance at 660 nm. Respectively 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 absorbances of the samples measured at 660 nm are A_sample, A_sample control, A_blank, and A_blank control, respectively, and calculate the inhibition rate according to the following formula. In addition, set acarbose (1 μg / μL) 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%.

[0117] 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 reduce blood glucose by inhibiting the activity of α-amylase.

[0118] Example 10

[0119] Cholesterol solution: 0.1 g of cholesterol, 0.2 g of bile salt, 0.11 g of sucrose ester, 1 mL of Tween 80, 5 mL of absolute ethanol, add to a 15 mL centrifuge tube. Ultrasonic treatment, after a hot water bath at 95 ± 5 °C, ultrasonic at 80 W for 20 min, vortex and mix evenly at intervals of 4 - 8 min until all cholesterol is dissolved, and filter and sterilize while it is hot (membrane pore size is 0.22 μm).

[0120] Add 360 μL of the above cholesterol solution to 30 mL of MRS liquid medium while it is still hot, and stir magnetically at 800 rpm for 15 min. Inoculate Lactobacillus acidophilus CCNH655 and LGG glycerol tubes into MRS liquid medium, normalize the OD, and transfer 2 vol% to the above MRS liquid medium containing cholesterol. Incubate at 37 °C and 180 rpm for 24 h, using the blank medium as a blank control. Use the cholesterol detection kit TC-W48-N (1031) to detect the cholesterol content in the supernatant of the fermentation broth at 500 nm. Calculate the cholesterol degradation rate according to the following formula: Degradation rate = (A 空白对照 -A 样品 ) / A 空白对照 × 100%.

[0121] The calculated cholesterol clearance rates are 79.39% and 34.06% respectively, indicating that the cholesterol degradation ability of Lactobacillus acidophilus CCNH655 is stronger than that of LGG.

[0122] Example 11

[0123] Inoculate Escherichia coli (CICC 10421 and ATCC 25922), Staphylococcus aureus (CMCC(B)26001 and CMCC(B)26003), Salmonella (ATCC 14028) and CVCC 3378) into LB liquid medium respectively, and inoculate Candida albicans (CICC 1965 and ATCC 10231) into YPD solid medium. Incubate aerobically at 37 °C for 18 h for activation, and then transfer to the corresponding fresh medium and culture until the viable cell count reaches 10 5 -10 7 CFU / mL, and dilute with the corresponding medium to 10 5 CFU / mL as the indicator bacterial solution. After centrifuging the expanded bacterial solution, take the supernatant and mix it with the bacterial solution of 10 5 CFU / mL pathogenic bacteria at a volume ratio of 1:1. Replace the supernatant with an equal volume of sterile MRS liquid medium as the negative control group, and replace the fermentation supernatant with 10 mL of kanamycin (content: 100 μg / mL) as the positive control group. Measure the OD 600 values of the experimental group and the negative control group, which are A and A 0， respectively. Calculate the inhibition rate according to the following formula, and the results are shown in Table 4.

[0124] Inhibition rate (%) = (A0 - A) / A0 × 100%.

[0125] Table 4

[0126]

[0127] The results showed that Lactobacillus acidophilus CCNH655 had certain inhibitory ability against Escherichia coli, Staphylococcus aureus and Salmonella.

[0128] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited thereto. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, including any other suitable combination of each technical feature. These simple modifications and combinations should also be regarded as the content disclosed by the present invention and fall within 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 preventing and / or treating hyperlipidemia.

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

8. The use according to any one of claims 5 to 7, wherein: The medicament may also contain corn oil and / or glycerin.

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