CN114292795B

CN114292795B - Lactobacillus probiotics CGMCC No.1.13855 and application thereof in preparation of lipid-lowering drugs

Info

Publication number: CN114292795B
Application number: CN202210068351.0A
Authority: CN
Inventors: 徐建国; 刘丽云; 杨晶; 任志鸿
Original assignee: National Institute for Communicable Disease Control and Prevention of Chinese Center For Disease Control and Prevention
Current assignee: National Institute for Communicable Disease Control and Prevention of Chinese Center For Disease Control and Prevention
Priority date: 2022-01-20
Filing date: 2022-01-20
Publication date: 2023-07-25
Anticipated expiration: 2042-01-20
Also published as: CN114292795A

Abstract

The invention discloses a lactobacillus probiotic strain, the preservation number of the strain is CGMCC No.1.13855, the preservation date is 2021, 12 months and 16 days, the preservation classification is Xu Jianguo lactobacillus (Lactobacillus xujianguonis), and the preservation unit is China general microbiological culture Collection center (China Committee for culture Collection). The invention also discloses application of the strain in preparation of lipid-lowering foods and health-care products, and the strain is harmless to animals and has the efficacy of lowering blood lipid.

Description

Lactobacillus probiotics CGMCC No.1.13855 and application thereof in preparation of lipid-lowering drugs

Technical Field

The invention relates to probiotics and application thereof, and belongs to the field of microorganisms.

Background

Lactobacillus belongs to lactic acid bacteria, and is named because the lactobacillus ferments carbohydrate to generate lactic acid products, and the appearance of the lactobacillus is in a short rod shape. The Lactobacillus is a genus of bacteria belonging to the genus Lactobacillus of the order Lactobacillus, belonging to the genus Lactobacillus of the genus Thick-walled bacteria. The first strain of Lactobacillus delbrueckii (L.delbrueckii subsp. Delbrueckii) of this genus was isolated from fermented cereal by Leichmann in 1896. The genus 2019-2020, which has a total of 48 new species, is now composed of 238 species. Lactobacillus is widely distributed in nature, and can be separated from plants, animals such as pigs, foods such as beer and pickled Chinese cabbage. Members of the genus lactobacillus are gram-positive bacteria, do not produce spores, have low tolerance to oxygen, are mostly anaerobic and unpowered, and are partly microaerophilic and powered. Under the condition of strict fermentation, lactobacillus can ferment from carbohydrate to produce lactic acid as main final product, so that the strain has better acid tolerance and partial strain has acidophilic property. Because of this property of lactobacillus, they are often used in the food fermentation industry, such as in the manufacture of cheese, yogurt, and the like. In addition, lactobacillus is considered to be a non-pathogenic and food-grade safe genus, and many strains play an important role in food microbiology and human nutrition, and are used as probiotics.

Existing studies indicate that lactobacillus has the beneficial effects on host health of regulating various biochemical reactions and immune responses of the host, such as antagonizing food-borne pathogenic bacteria by producing antibacterial complexes and antibiotics; reducing the abundance level of pathogenic and opportunistic pathogens by competitively adhering host cells to the pathogen; regulating host immune response by cell regulators such as teichoic acid; the cholesterol absorption rate and serum cholesterol level are reduced by the hydrolysis of bile salts and the absorption of cholesterol, so that the cholesterol absorption rate and serum cholesterol level are reduced, and the cholesterol absorption rate and serum cholesterol level are reduced.

Hyperlipidemia in humans can directly cause diseases that are serious harm to human health, such as atherosclerosis, coronary heart disease, pancreatitis, etc. Currently, hyperlipidemia is mainly reduced by diet and exercise to control body weight, while medication is being performed, including statins and resins, which are mainly lowering serum total cholesterol and LDL cholesterol, and fibrates and nicotinic acids, which are mainly lowering serum triacylglycerols. Although lipid-lowering drugs can function to lower blood lipid, side effects of drugs are increasingly being focused by clinicians, probiotics, particularly food-grade lactic acid bacteria, are increasingly being recognized by researchers as lipid-lowering products, and lipid-lowering efficacy of some lactic acid bacteria is being researched and developed.

The invention aims to separate and screen novel lactobacillus species with good cholesterol-reducing function from the excrement of Qinghai wild woodchuck which only ingests plants, and further develop the application of the novel lactobacillus species in preparing lipid-lowering medicaments.

Disclosure of Invention

Based on the above objects, the present invention provides a lactobacillus probiotic strain, wherein the preservation number of the probiotic strain is CGMCC No.1.13855, the preservation date is 2021, 12 and 16 days, the preservation classification is Xu Jianguo lactobacillus (Lactobacillus xujianguonis), the preservation unit is China general microbiological culture Collection center, and the address is post code 100101 of national institute of Chinese academy of sciences, north Chen West road 1, beijing, the Korean region.

In a preferred embodiment, the 16S rDNA of the strain has the sequence shown in SEQ ID NO: 1.

The invention further provides application of the strain in preparation of lipid-lowering medicaments.

In a preferred embodiment, the lipid-lowering drug is a drug that lowers human serum total cholesterol, triglycerides, low density lipoproteins, and/or raises high density lipoproteins.

In another preferred embodiment, the lipid-lowering drug is a drug that reduces the fat content of the liver.

In a preferred embodiment of the use of the strain described above in the preparation of a lipid-lowering medicament, the lipid-lowering medicament may also be used as a medicament for the treatment of liver diseases. The liver disease may alternatively be liver dysfunction due to lipid metabolism disorders, or hepatitis and/or liver fibrosis caused by inflammatory factors including, but not limited to, interleukin 1-beta, serum monocyte chemotactic protein-1, and the like.

In a preferred embodiment of the use of the strain described above for the preparation of a lipid-lowering medicament, the lipid-lowering medicament may also be used as a medicament for the treatment of diabetes. The diabetes treatment agent may alternatively be an agent that reduces the glucose level in human serum, or an agent that reduces human serum insulin, or an agent that reduces inflammatory factors in human serum that are associated with insulin resistance, including, but not limited to, tumor necrosis factor-alpha and/or IL-6.

Finally, the invention also provides a composition of the strain.

In a preferred embodiment, the composition is prepared as a capsule, a lyophilized powder or a bacterial liquid formulation.

The invention separates and purifies lactobacillus with probiotic property from wild woodchuck faeces, and experiments prove that the separated lactobacillus is harmless to animals, and the lactobacillus has the efficacy of regulating blood fat through animal experiments. The CGMCC 1.13855 strain can obviously reduce low-density lipoprotein, total cholesterol and triglyceride, raise the function of high-density lipoprotein and has similar blood fat regulating effect with simvastatin. The physical sign of the rat which ingests the lactobacillus is normal, and the gastrointestinal liver and spleen are taken as pathology after the experiment is finished after five weeks of ingestion, so that the abnormality is not seen. The bacteria detection of the aseptic tissue does not see the ectopic field planting of the lactobacillus.

Drawings

FIG. 1.CGMCC No.1.13855 strain is based on phylogenetic tree analysis of 16S rDNA genes (maximum likelihood method);

FIG. 2.CGMCC No.1.13855 effect of strain on serum Total Cholesterol (TC) in high fat model rats;

FIG. 3.CGMCC No.1.13855 effect of strain on serum Triglycerides (TG) of high-fat model rats;

FIG. 4.CGMCC No.1.13855 effect of strain on serum Low Density Lipoprotein (LDL) in high fat model rats;

FIG. 5.CGMCC No.1.13855 effect of strain on serum High Density Lipoprotein (HDL) of high fat model rats;

FIG. 6.CGMCC No.1.13855 effect of strain on liver fat droplet content of high fat model rats (oil red O staining);

FIG. 7.CGMCC No.1.13855 effect of strain on serum glutamic pyruvic transaminase (ALT) of high-fat model rats;

FIG. 8.CGMCC No.1.13855 effect of strain on serum glutamate oxaloacetic transaminase (AST) of high-fat model rats;

FIG. 9.CGMCC No.1.13855 effect of strain on hepatic Malondialdehyde (MDA) in high-fat model rats;

FIG. 10.CGMCC No.1.13855 effect of strain on liver superoxide dismutase (SOD) in high lipid model rats;

FIG. 11.CGMCC No.1.13855 effect of strain on liver glutathione peroxidase (GSH-Px) in high fat model rats;

FIG. 12.CGMCC No.1.13855 effect of strain on serum interleukin 1-beta (IL-1 beta) in high-fat model rats;

FIG. 13.CGMCC No.1.13855 effect of strain on serum monocyte chemotactic protein-1 (MCP-1) of high-fat model rats;

FIG. 14.CGMCC No.1.13855 effect of strain on serum Endotoxin (ET) of high fat model rats;

FIG. 15.CGMCC No.1.13855 effect of strain on fasting blood glucose in high lipid model rats;

FIG. 16.CGMCC No.1.13855 effect of strain on fasting serum insulin in high fat model rats;

FIG. 17.CGMCC No.1.13855 effect of strain on serum tumor necrosis factor- α (TNF- α) in high fat model rats;

FIG. 18.CGMCC No.1.13855 effect of strain on serum interleukin 6 (IL-6) in high fat model rats.

Detailed Description

The invention will be further described with reference to specific embodiments, and advantages and features of the invention will become apparent from the description. These examples are merely exemplary and do not limit the scope of the invention in any way.

EXAMPLE 1 isolation, screening and identification of lactic acid bacteria

1. Isolation of lactic acid bacteria

1) Taking 100 μl of sample from the bacteria-retaining tube, adding into EP tube preloaded with 900 μl of sterile PBS, sequentially performing gradient dilution on the sample, and diluting the woodchuck faeces sample to 10 ^-6 Doubling;

2) 100 mu L of samples with different dilutions are coated on MRS culture medium and put into an incubator;

3) At 37℃0.5% CO ₂ Culturing in the environment for 48h;

4) Taking out the culture dish, picking colonies with different forms of characteristics by using a sterile inoculating loop, transferring to a new MRS solid culture medium for purification, performing anaerobic culture at 37 ℃ for 48 hours, continuously transferring for 3 times, culturing the purified strain in liquid MRS with pH of=3.5, and screening strains with excellent acid resistance growth for experiment or freezing preservation.

2. Preservation of bacterial species

The laboratory uses MRS culture medium containing 25% glycerol as bacteria-preserving liquid to carry out the freezing preservation of strains, and the method is as follows:

1) Sterilizing 2mL of bacteria-retaining tube at 121deg.C for 15 min;

2) After lactobacillus is continuously transferred on MRS solid culture medium for 3 times, 1.5ml of sterile bacteria-preserving liquid is added on a culture dish;

3) Scraping and coating the culture dish by using an L rod to fully integrate bacterial colonies into the bacteria-retaining liquid;

4) Transferring the bacterial liquid into a bacteria-preserving tube, uniformly mixing and preserving at-80 ℃.

3. Colony appearance and bacterial morphology observations

Lactobacillus HT111-2 with the preservation number of CGMCC No.1.13855 belongs to anaerobic bacteria, grows well under anaerobic conditions, forms light yellow colonies with round shape, irregular edge, raised middle and rough surface on a Columbia blood culture medium, and has the diameter of about 1-1.2mm; does not grow under aerobic conditions. Under an optical microscope, the thalli are light purple after gram staining, and can be arranged end to end. The thallus is rod-shaped under a transmission electron microscope, has no spore, no flagellum, no bacteria, no power, and has a diameter of about 0.7-1×2.5-4.28 μm.

4. Extraction of total DNA from bacteria

Single colonies were inoculated on MRS medium, cultured overnight at 37℃and DNA was extracted according to the bacterial genomic DNA extraction kit (TIANGEN) protocol.

5. Biochemical identification method of strain

The present study uses the "BioMerieux" bacterial System Biochemical identification cards API 50CH and API ZYM, manufactured by Merieux, france, to perform carbohydrate glycolysis and enzymatic biochemical identification of strain HT111-2 as well as of the reference strain.

Results: according to biochemical identification, a strain with biochemical characteristics of lactobacillus is obtained, the strain preservation number is CGMCC No.1.13855, the preservation date is 2021, 12 months and 16 days, the preservation classification is named Xu Jianguo lactobacillus (Lactobacillus xujianguonis), the preservation unit is China general microbiological culture Collection center, and the address is post code 100101 of China academy of sciences of national academy of sciences of North Chen West road 1, beijing, korea, and the preservation classification is Xu Jianguo lactobacillus.

6. Bacterial universal primer 16S rRNA PCR amplification

Identification of bacterial 16S rRNA: extracting bacterial genome DNA, amplifying a lactobacillus universal primer 16S rDNA PCR product, sequencing, and performing BLAST comparison on the sequence on NCBI for preliminary identification.

The primers for PCR amplification of bacterial 16S rRNA and the PCR reaction conditions used in this experiment were as follows, and 50. Mu.L of PCR system was used for all experiments. And (3) carrying out BLAST comparison on the sequencing result of the lactobacillus universal primer 16S rRNA PCR product to be tested on NCBI, and identifying. The result suggests that the CGMCC No.1.13855 strain is a suspected new species of lactobacillus.

Universal primer 16S rRNA PCR amplification conditions

Primer sequence:

27F，5′-AGAGTTTGATCMTGGCTCAG-3′；

1492R 5′-TACGGYTACCTTGTTACGACTT-3′；

reaction system (50 μl):

Premix Taq	25μl
		dd H ₂ O	19μl
27F/1492R	2/2μl
		DNA	2μl

；

amplification conditions:

7. phylogenetic analysis

According to the strain CGMCC No.1.1385516S rDNA GenBank 31 model strain 16S rDNA gene as a reference sequence, using an Enterococcus ATCC 19434 16S rDNA gene sequence as an outer group, using MEGA 6.0 software, adopting CLUSTAL_W to carry out multi-sequence alignment, and constructing a system evolution tree based on a maximum likelihood method MaximumLikelihood (ML). The phylogenetic tree constructed based on the 16S rDNA gene sequence shows that the strain CGMCC No.1.13855 is clustered with adjacent lactobacillus, and an independent branch is formed on the phylogenetic tree, which is most similar to the evolutionary relationship of L.amylolyticus DSM 11664 and L.armteri DSM 5661, and is shown in the figure 1 (maximum likelihood method). The full genome frame map sequence of the strain CGMCC No.1.13855 is obtained by second generation sequencing, and hybridized with the genome of the reference strain L.amylolyticus DSM 11664 and L.hamsteri DSM 5661 on line, the DDH values are respectively 20.6 percent and 21.1 percent, and the DDH values are respectively hybridized with the genome sequences of 175 mode strains in the lactobacillus genus recorded in a GenBank database on line, and the result shows that the DDH value range is 17.2-42.5 percent and is lower than 70 percent, thereby meeting the judging standard of new species.

In conclusion, genome and phylogenetic analysis suggest that the strain CGMCC No.1.13855 is a novel species of Lactobacillus.

Example 2 hypolipidemic function of Lactobacillus CGMCC No.1.13855 on rat serum

32 female SD rats of about 200 g body weight were randomly divided into four groups, one group was given normal maintenance feed, and three groups were given high fat feed (cholesterol 1%, lard 10%,0.2 cholate, 10% yolk powder). Three groups of high fat diet groups were orally administered 10 a week after one week on alternate days ⁹ CFU CGMCC No.1.13855 lactobacillus solution, physiological saline solution and simvastatin with weight of 20mg/kg are administered every three weeks before the termination of the experimentThe experiment is terminated for five weeks, and the blood lipid level of serum is detected to evaluate the preventive intervention effect of lactobacillus CGMCC No.1.13855 on the lipid metabolism disorder model.

FIG. 2 shows CGMCC No.1.13855 ^T Effects on serum Total Cholesterol (TC) in high-fat model rats. The average content of serum TC in CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and simvastatin intervention group is 1.79+/-0.18 mmol/L and 1.77+/-0.08 mmol/L which are obviously lower than that in high-fat model group (p)<0.001 A) is provided; the average content of serum TC in the high-fat model group is 2.48+/-0.23 mmol/L, which is obviously higher than that in the normal control group by 1.40+/-0.10 mmol/L (p)<0.001)。

FIG. 3 shows the effect of CGMCC No.1.13855 on serum Triglyceride (TG) of a high-fat model rat. The average content of serum TG in the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 2.55+/-0.17 mmol/L and 2.55+/-0.08 mmol/L, which are obviously lower than that in the high-fat model group (p < 0.001); the average content of serum TG in the high-fat model group is 3.07+/-0.15 mmol/L, which is obviously higher than that in the normal control group by 1.87+/-0.09 mmol/L (p < 0.001).

FIG. 4 shows the effect of CGMCC No.1.13855 on serum Low Density Lipoprotein (LDL) of high fat model rats. The average content of serum LDL in the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 2.60+/-0.15 mmol/L and 2.17+/-0.32 mmol/L, which are obviously lower than that of the high-fat model group (p <0.01 and p < 0.001); the average content of serum LDL in the high-fat model group is 3.15+/-0.24 mmol/L, which is obviously higher than that in the normal control group by 1.79+/-0.26 mmol/L (p < 0.001).

FIG. 5 shows the effect of CGMCC No.1.13855 on serum High Density Lipoprotein (HDL) of a high lipid model rat. The average serum HDL content of the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 2.10 plus or minus 0.22mmol/L and 2.33 plus or minus 0.17mmol/L, which are obviously higher than that of the high-fat model group by 1.51 plus or minus 0.08mmol/L (p <0.01 and p < 0.001); the average serum HDL content of the high-fat model group is 1.51+/-0.08 mmol/L, which is obviously lower than that of the normal control group by 3.10+/-0.41 mmol/L (p < 0.001).

FIG. 6 shows the effect of CGMCC No.1.13855 strain on liver fat droplet content (oil red O staining) of high fat model rats. Normal control oil red O staining showed no lipid droplets in the cytoplasm. The high-fat model group oil red O staining showed a large number of lipid droplets within the cytoplasm. The CGMCC No.1.13855 intervention group (i.e. HT111-2 intervention group) and simvastatin intervention group were stained with oil red O showing lipid droplets in the cytoplasm, but significantly less than the high-lipid model group.

The results show that the CGMCC No.1.13855 lactobacillus strain has the functions of obviously reducing serum total cholesterol, triglyceride, low density lipoprotein and increasing high density lipoprotein, can reduce the fat content of liver, and has similar blood fat reducing effect as simvastatin.

Rats ingested with CGMCC No.1.13855 lactobacillus appeared normally, and no abnormality was seen in taking gastrointestinal liver and spleen as pathology after five weeks of ingestion ended the experiment. The bacteria detection of the aseptic tissue does not see the ectopic field planting of the lactobacillus.

Example 3. Protective effect of lactobacillus CGMCC No.1.13855 on liver function of rats 32 female SD rats weighing about 200 grams were randomly divided into four groups, one group being given normal maintenance feed and three groups being given high fat feed (cholesterol 1%, lard 10%,0.2 cholate, 10% egg yolk powder). Three groups of high fat diet groups were orally administered 10 a week after one week on alternate days ⁹ The CFU CGMCC 1.13855 lactobacillus solution is administered with physiological saline at intervals, and with simvastatin with the weight of 20mg/kg at intervals three weeks before the termination of the experiment, the experiment is terminated five weeks in total, and the levels of glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase and inflammatory factors of serum are detected to evaluate the preventive intervention effect of the lactobacillus CGMCC No.1.13855 on liver function injury caused by lipid metabolism disorder models.

FIG. 7 shows the effect of CGMCC No.1.13855 on serum glutamic pyruvic transaminase (ALT) of a high-fat model rat. The average serum ALT content of the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 1.05+/-0.09 ng/mL and 1.34+/-0.04 ng/mL, which are obviously lower than that of the high-fat model group (p < 0.001); serum ALT average content of the high-fat model group is 1.66+/-0.09 ng/mL, which is obviously higher than that of the normal control group by 0.94+/-0.05 ng/mL (p < 0.001).

FIG. 8 shows the effect of CGMCC No.1.13855 on serum glutamic-oxaloacetic transaminase (AST) of a high-fat model rat. The average serum AST content of the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 1.67+/-0.26 ng/mL and 1.91+/-0.34 ng/mL, which are obviously lower than that of the high-fat model group (p < 0.001); serum AST average content of the high-fat model group is 2.70+/-0.12 ng/mL, which is obviously higher than that of the normal control group by 1.51+/-0.27 ng/mL (p < 0.001).

Figure 9 shows the effect of CGMCC No.1.13855 on hepatic Malondialdehyde (MDA) in high lipid model rats. The average content of liver MDA in CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and simvastatin intervention group is 3.12 plus or minus 0.38nmol/mL and 3.45 plus or minus 0.49nmol/mL, which are obviously lower than that of high-fat model group 4.54 plus or minus 0.20nmol/mL (p <0.01 and p < 0.001); the average content of liver MDA in the high-fat model group is 4.54+/-0.20 nmol/mL, which is obviously higher than that in the normal control group by 2.63+/-0.49 nmol/mL (p < 0.001).

FIG. 10 shows the effect of CGMCC No.1.13855 on liver superoxide dismutase (SOD) of a high-lipid model rat. The average content of liver SOD in CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and simvastatin intervention group is 1.28+/-0.07 ng/mL and 1.18+/-0.14 ng/mL, which are both obviously higher than that in high-fat model group by 0.90+/-0.05 ng/mL (p < 0.001); the average liver SOD content of the high-fat model group is 0.90+/-0.05 ng/mL, which is obviously lower than that of the normal control group by 1.32+/-0.06 ng/mL (p < 0.001).

FIG. 11 shows the effect of CGMCC No.1.13855 on liver glutathione peroxidase (GSH-Px) of a high fat model rat. The average content of liver GSH-Px of CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and simvastatin intervention group is 7.71+/-1.08 ng/mL and 7.80+/-0.63 ng/mL, which are obviously higher than that of high-fat model group 4.31+/-0.41 ng/mL (p < 0.001); the average liver GSH-Px content of the high fat model group is 4.31+/-0.41 ng/mL, which is obviously lower than that of the normal control group by 8.90+/-0.48 ng/mL (p < 0.001).

The results show that the CGMCC No.1.13855 lactobacillus strain has the effects of obviously reducing serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase and liver malondialdehyde, and simultaneously increases the effects of liver superoxide dismutase and liver glutathione peroxidase, and has the similar effect of protecting liver function with simvastatin.

EXAMPLE 4 liver inflammation and liver fibrosis-related inflammatory factor reduction function of Lactobacillus CGMCC No.1.13855 on rat serum

FIG. 12 shows the effect of CGMCC No.1.13855 on serum interleukin 1-beta (IL-1 beta) of a high-fat model rat. The average content of serum IL-1 beta in the CGMCC 1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 20.29+/-1.88 pg/mL and 17.98+/-1.16 pg/mL, which are obviously lower than that in the high-fat model group (p < 0.001); the average content of serum IL-1 beta in the high-fat model group is 28.40+/-1.96 pg/mL, which is obviously higher than that in the normal control group, namely 20.06+/-3.81 pg/mL (p < 0.001).

FIG. 13 shows the effect of CGMCC No.1.13855 on serum monocyte chemotactic protein-1 (MCP-1) of a high-fat model rat. The average content of serum MCP-1 in the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) is 536.98 +/-104.31 pg/mL, which is obviously lower than 733.69 +/-40.18 pg/mL (p < 0.001) of the high-fat model group; the average content of serum MCP-1 in the simvastatin intervention group is 628.89 +/-52.98 pg/mL, and has no obvious difference (p is more than 0.05) with 733.69 +/-40.18 pg/mL of the high-fat model group; the average serum MCP-1 content of the high-fat model group is 733.69 +/-40.18 pg/mL, which is obviously higher than that of the normal control group 527.28 +/-72.42 pg/mL (p < 0.001).

The results show that the CGMCC No.1.13855 lactobacillus strain has the effect of obviously reducing serum IL-1 beta and MCP-1 inflammatory factors and has stronger effect of reducing serum inflammatory factors than simvastatin. Liver fibrosis is the final outcome of the continued development of liver injury from a variety of causes, and MCP-l is an important macrophage chemokine in tissue injury repair, which plays an important role in liver injury and repair. IL-1β is another important inflammatory cell that causes fibrosis, is the primary inducer of the pro-inflammatory response, and can enhance the extent of the inflammatory response by coordinating and promoting the expression of other inflammatory factors, such as TNF- α. In summary, MCP-l and IL-1β are important inflammatory factors in liver fibrosis. The CGMCC No.1.13855 lactobacillus obviously reduces the content of serum IL-1 beta and MCP-1 inflammatory factors, thereby reducing the occurrence of liver inflammation and liver fibrosis.

EXAMPLE 5 endotoxin-reducing function of Lactobacillus CGMCC No.1.13855 on rat serum

Figure 14 shows the effect of CGMCC 1.13855 on serum Endotoxin (ET) of high fat model rats. The average content of serum ET in CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) is 36.14 +/-5.59 pg/mL, which is obviously lower than 53.95+/-6.28 pg/mL (p < 0.001) of high-fat model group; the average serum ET content of the simvastatin intervention group is 44.89+/-4.90 pg/mL, and has no obvious difference (p is more than 0.05) with 53.95+/-6.28 pg/mL of the high-fat model group; the average serum ET content of the high fat model group is 53.95+/-6.28 pg/mL, which is obviously higher than that of the normal control group which is 25.25+/-6.26 pg/mL (p < 0.001).

The results show that the CGMCC No.1.13855 lactobacillus strain has the effect of obviously reducing serum Endotoxin (ET), thereby reducing the liver injury degree and having stronger effect of reducing the serum Endotoxin (ET) than simvastatin.

EXAMPLE 6 in vivo evaluation of hypoglycemic Functions of Lactobacillus CGMCC No.1.13855

32 female SD rats of about 200 g body weight were randomly divided into four groups, one group was given normal maintenance feed, and three groups were given high fat feed (cholesterol 1%, lard 10%,0.2 cholate, 10% yolk powder). Three groups of high fat diet groups were orally administered 10 a week after one week on alternate days ⁹ The CFU CGMCC No.1.13855 lactobacillus liquid is administered with physiological saline every other day, and with simvastatin with the weight of 20mg/kg every other day three weeks before the termination of the experiment, and the blood sugar and inflammatory factor level of the fasting serum of the rat are detected to evaluate the preventive intervention effect of the lactobacillus CGMCC No.1.13855 on the metabolic disorder model.

1. The lactobacillus CGMCC No.1.13855 has the function of reducing fasting blood glucose and blood insulin of rats

Figure 15 shows the effect of CGMCC No.1.13855 on fasting blood glucose in a high-fat model rat. The average blood sugar content of the CGMCC 1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 3.30+/-0.78 mmol/L and 2.91+/-0.72 mmol/L, which are obviously lower than that of the high-fat model group (p < 0.001); the average blood sugar content of the high-fat model group is 5.09+/-0.29 mmol/L, which is obviously higher than that of the normal control group by 2.67+/-0.41 mmol/L (p < 0.001).

Figure 16 shows the effect of CGMCC No.1.13855 on fasting serum insulin in a high-fat model rat. The average serum insulin contents of the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group are 31.45 +/-1.77 mU/L and 36.81+/-2.23 mU/L which are obviously lower than that of the high-fat model group 44.84+/-2.23 mU/L (p < 0.001); the average serum insulin content of the high-fat model group is 44.84+/-2.23 mU/L, which is obviously higher than that of the normal control group by 25.33+/-3.04 mU/L (p < 0.001).

The results show that the CGMCC No.1.13855 lactobacillus strain has the effect of obviously reducing blood sugar and serum insulin and has similar blood sugar reducing effect as simvastatin. Lipid metabolism disorders such as hypertriglyceridemia and hyperfree fatty acidemia are closely related to insulin resistance. Insulin resistance reduces the efficiency of insulin in promoting glucose uptake and utilization, and the body's compensatory hypersecretion of insulin produces hyperinsulinemia to maintain blood glucose stability. Insulin resistance is prone to metabolic syndrome and type 2 diabetes. The lactobacillus strain CGMCC No.1.13855 has the function of reducing blood fat, thereby reducing insulin resistance and leading to the reduction of blood sugar and serum insulin.

EXAMPLE 7 insulin resistance action-related inflammatory factor reducing function of Lactobacillus CGMCC No.1.13855 on rat serum

FIG. 17 shows the effect of CGMCC No.1.13855 on serum tumor necrosis factor-alpha (TNF-alpha) of a high-fat model rat. The average content of serum TNF-alpha in the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 112.54 +/-20.53 pg/mL and 159.75 +/-7.95 pg/mL, which are obviously lower than 198.5+/-13.94 pg/mL (p < 0.001) of the high-fat model group; the average content of the serum TNF-alpha in the high-fat model group is 198.5+/-13.94 pg/mL, which is obviously higher than that in the normal control group 105.44 +/-25.22 pg/mL (p < 0.001).

FIG. 18 shows the effect of CGMCC No.1.13855 on serum interleukin 6 (IL-6) of a high-fat model rat. The average serum IL-6 content of the CGMCC No.1.13855 intervention group (namely HT111-2 intervention group) and the simvastatin intervention group is 115.43 +/-9.20 pg/mL and 132.24 +/-9.94 pg/mL, which are obviously lower than 155.93 +/-7.55 pg/mL (p <0.001 and p < 0.01) of the high-fat model group; the average content of serum IL-6 in the high-fat model group is 155.93 +/-7.55 pg/mL, which is obviously higher than 114.42 +/-8.90 pg/mL (p < 0.001) of the normal control group.

The results show that the CGMCC No.1.13855 lactobacillus strain has the effects of obviously reducing serum tumor necrosis factor-alpha and IL-6 inflammatory factors and reducing serum inflammatory factors more than simvastatin. Because the production of insulin resistance is closely related to serum inflammatory factor levels, long-term over-secretion of serum inflammatory factors may be a major factor in impaired insulin secretion by islet beta cells and in the development of insulin resistance. TNF- α and IL-6, as the most representative cytokines, play a central regulatory role in inflammation and immune responses. Elevated serum levels of TNF- α and IL-6 can induce insulin resistance by interfering with insulin signaling. The CGMCC No.1.13855 lactobacillus strain obviously reduces the content of inflammatory factors of TNF-alpha and IL-6 in serum, thereby reducing the insulin resistance effect and causing the blood sugar to be reduced.

Sequence listing

<110> infectious disease prevention and control institute of China center for disease prevention and control

<120> lactobacillus probiotic CGMCC No.1.13855 and application thereof in preparation of lipid-lowering drugs

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1527

<212> DNA

<213> Lactobacillus xujianguonis

<400> 1

agagtttgat catggctcag gacgaacgct ggcggcgtgc ctaatacatg caagtcgagc 60

gagcagaact aacagattta cttcggtaat gacgtttcgg acgcgagcgg cggatgggtg 120

agtaacacgt gggtaacctg cccttaagtc tgggatacca cttggaaaca ggtgctaata 180

ccggataaca actagtgctg catggcacta gcttaaaagg cggcgtaagc tgtcgctaaa 240

ggatggaccc gcggtgcatt agctagttgg taaggtaacg gcttaccaag gcgacgatgc 300

atagccgagt tgagagactg atcggccaca ttgggactga gacacggccc aaactcctac 360

gggaggcagc agtagggaat cttccacaat gggcgaaagc ctgatggagc aacgccgcgt 420

gagtgaagaa ggttttcgga tcgtaaagct ctgttgttgg tgaagaagga tagaggtagt 480

aactggcctt tatttgacgg taatcaacca gaaagtcacg gctaactacg tgccagcagc 540

cgcggtaata cgtaggtggc aagcgttgtc cggatttatt gggcgtaaag cgagcgcagg 600

cggagaaata agtctgatgt gaaagccctc ggcttaaccg aggaagtgca tcagaactgt 660

ttttcttgag tcagaagagg agagtgaact ccatgtgtag cggtggaatg cgtagatata 720

tggaagaaca ccagtggcga aggcggctct ctggtctgta actgacgctg aggctcgaaa 780

gcatgggtag cgaacaggat tagataccct ggtagtccat gccgtaaacg atgagtgcta 840

agtgttggga ggtttccgcc tctcagtgct gcagctaacg cattaagcac tccgcctggg 900

gagtacgacc gcaaggttga aactcaaagg aattgacggg ggcccgcaca agcggtggag 960

catgtggttt aattcgaagc aacgcgaaga accttaccag gtcttgacat ctggtgcaaa 1020

cctaagagat taggcgttcc cttcggggac accaagacag gtggtgcatg gctgtcgtca 1080

gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg ttattagttg 1140

ccagcattaa gttgggcact ctaatgagac tgccggtgac aaaccggagg aaggtgggga 1200

cggcgtcaag tcatcatgcc ccttatgacc tgggctacac acgtgctaca atgggcagta 1260

caacgaggag cgaacctgtg aaggcaagcg aatctctgaa agctgttctc agttcggact 1320

gtaggctgca actcgcctac acgaagctgg aatcgctagt aatcgcggat cagcacgccg 1380

cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatggaa gtctgcaatg 1440

cccaaagccg gtggcctaac cttcgggaag gagccgtcta aggcagggca gatgactggg 1500

gtgaagtcgt aacaaggtaa ccgtaaa 1527

Claims

1. The application of lactobacillus probiotic strain in preparing lipid-lowering medicament is characterized in that the lactobacillus probiotic strain has a preservation number of CGMCC No.1.13855, a preservation date of 2021 and 12 months and 16 days, and a preservation classification of Xu Jianguo lactobacillus @ is namedLactobacillus xujianguonis) The preservation unit is China general microbiological culture Collection center.

2. The use according to claim 1, wherein the lipid lowering drug is a drug that lowers human serum total cholesterol, triglycerides, low density lipoproteins, and/or raises high density lipoproteins.

3. The use according to claim 1, wherein the lipid-lowering drug is a drug that reduces the fat content of the liver.

4. The use according to claim 1, wherein the lipid lowering drug is also a therapeutic drug for liver diseases caused by lipid metabolism disorders.

5. The use according to claim 1, wherein the lipid-lowering drug is simultaneously a diabetes therapeutic drug.