A strain of animal-derived Lactobacillus LA-1, microbial preparations and their applications

Abstract

The present invention relates to the field of microbial technology, and in particular to a strain of animal-associated Lactobacillus LA-1, a microbial preparation and an application thereof. Animal Lactobacillus ) LA‑1 has been deposited in the General Microbiology Center of China National Committee for the Preservation of Microbiological Cultures on September 27, 2024, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 32098. Animal-associated Lactobacillus LA‑1 has the effect of significantly reducing cholesterol content and bile salt hydrolase activity, and can inhibit α-glucosidase activity, has a high fatty acid absorption rate, is safe and sensitive to commonly used antibiotics, can relieve obesity, reduce blood lipids and liver lipid accumulation, relieve liver lipid degeneration and fat hypertrophy, has the effect of protecting the liver and enhancing liver function, and has good application prospects in the preparation of functional products.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to an animal-associated lactobacillus LA-1, a microbial preparation and applications thereof. Background Art

[0002] Lipid metabolism plays a vital role in maintaining animal life and ensuring normal physiological functions. Lipid metabolism disorders in overweight or obese people can easily cause liver damage, such as metabolic associated fatty liver disease (MAFLD), which can further develop into hepatitis, cirrhosis, and related physiological diseases such as hyperlipidemia, cardiovascular disease, diabetes, etc.

[0003] Statins are currently commonly used to treat hypercholesterolemia, but these drugs have certain side effects on the human body. Therefore, seeking safe and effective natural substances for preventing or treating lipid metabolism disorders is a hot topic of research. Probiotics are safe and have no toxic side effects. They can affect the intestinal flora and intestinal flora products through the regulation of intestinal flora, and then have a beneficial effect on the body. Chinese invention patent CN116970539B discloses a mouse combined lactobacillus, which has the effect of lowering blood lipids or inhibiting weight gain caused by high fat. However, there are large differences between strains. If more probiotics with the effects of alleviating obesity and lowering cholesterol can be discovered, it will have important research value and application prospects for preventing or treating lipid metabolism disorders.

[0004] There is a lack of research on animal-associated lactobacilli with probiotic properties such as regulating lipid metabolism. Summary of the invention

[0005] In view of the technical problem that there is still a lack of research on animal-associated Lactobacillus with probiotic properties such as regulating lipid metabolism, the present invention provides an animal-associated Lactobacillus LA-1, a microbial preparation and an application thereof.

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

[0007] In a first aspect, the present invention provides an animal-associated Lactobacillus ( Ligilactobacillus animalis )LA-1, the strain was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on September 27, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.32098.

[0008] In a second aspect, the present invention provides a microbial preparation comprising the above-mentioned animal-associated Lactobacillus LA-1.

[0009] Furthermore, the microbial preparation is a liquid preparation or a solid preparation.

[0010] Furthermore, the content of animal combined lactobacillus LA-1 in the liquid preparation is 1.0×10 7 ~1.0×10 10 CFU / mL; the content of animal-associated Lactobacillus LA-1 in the solid preparation is 1.0×10 7 ~1.0×10 10 CFU / g.

[0011] In a third aspect, the present invention provides an application of the above-mentioned animal combined Lactobacillus LA-1 or the above-mentioned microbial preparation in the preparation of a functional product, and the functional product can be used for (1) preventing, alleviating or treating obesity; (2) preventing, alleviating or treating metabolic syndrome; (3) preventing, alleviating or treating hyperlipidemia.

[0012] Furthermore, animal-associated Lactobacillus LA-1 has the following functions:

[0013] (a) Reduce triglyceride content in the liver and serum; (b) Reduce total cholesterol content in the liver, reduce low-density lipoprotein cholesterol, and increase low-density lipoprotein cholesterol; (c) Reduce liver and serum alanine aminotransferase and aspartate aminotransferase levels, enhance liver function, and protect the liver; (d) Reduce serum lactate dehydrogenase; (e) Improve liver and fat pathological characteristics; (f) Regulate intestinal flora.

[0014] Furthermore, the functional product may be food or medicine.

[0015] The beneficial effects of the present invention are:

[0016] The present invention separates an animal-associated lactobacillus LA-1 from healthy dog ​​feces, which can proliferate in the gastrointestinal fluid environment, reach the gastrointestinal tract of animals to exert its effects, has the effect of significantly reducing cholesterol content and bile salt hydrolase activity, and can inhibit α-glucosidase activity, has a high fatty acid absorption rate, is safe and sensitive to commonly used antibiotics, relieves obesity, reduces blood lipids and liver lipid accumulation, relieves liver lipid degeneration and fat hypertrophy, has the effect of protecting the liver and enhancing liver function, and has good application prospects in the preparation of functional products (such as food, food additives, feed additives, medicines and health products for human or animal consumption or drinking). BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0018] Figure 1This is the colony morphology of animal-associated Lactobacillus LA-1.

[0019] Figure 2 This is a microscopic examination picture of animal-associated Lactobacillus LA-1.

[0020] Figure 3 It is a bar graph of bacterial survival rate of animal combined Lactobacillus LA-1 in Example 1 at different time periods in artificial gastrointestinal fluid.

[0021] Figure 4 These are photos of the results of the bile salt hydrolase activity test in Example 1, (A) is a photo of the filter paper on the MRS plate, (B) is a photo of the filter paper on the sodium glycocholate plate, and (C) is a photo of the filter paper on the sodium taurocholate plate.

[0022] Figure 5 These are photos of the results of the hemolytic test in Example 1, (A) is a photo of the results of the hemolytic test on animals and Lactobacillus LA-1, and (B) is a photo of the results of the hemolytic test on Escherichia coli K88.

[0023] Figure 6 2 is a graph showing the weight changes of mice in different treatment groups in Example 2.

[0024] Figure 7 It is a bar graph of organ indexes of mice in different treatment groups in Example 2.

[0025] Figure 8 This is a staining image of the liver tissue of mice in different treatment groups in Example 2.

[0026] Fig. 9 This is the staining image of the adipose tissue of mice in different treatment groups in Example 2.

[0027] Fig.10 The figures are histograms of serum indicators of mice in different treatment groups in Example 2, (A) is a histogram of serum total cholesterol, (B) is a histogram of serum triglycerides, (C) is a histogram of serum low-density lipoprotein cholesterol, (D) is a histogram of serum alanine aminotransferase, (E) is a histogram of serum aspartate aminotransferase, and (F) is a histogram of serum lactate dehydrogenase.

[0028] Fig.11 The figures are the bar graphs of liver indexes of mice in different treatment groups in Example 2, (A) is a bar graph of liver total cholesterol, (B) is a bar graph of liver triglycerides, (C) is a bar graph of liver high-density lipoprotein cholesterol, (D) is a bar graph of liver low-density lipoprotein cholesterol, (E) is a bar graph of liver very-low-density lipoprotein cholesterol, (F) is a bar graph of liver glucose, (G) is a bar graph of liver alanine aminotransferase, and (H) is a bar graph of liver aspartate aminotransferase.

[0029] Fig.12 This is the PCoA analysis diagram of the feces of mice in different treatment groups in Example 2.

[0030] Fig.13 It is a bar graph of the intestinal flora composition of mice in different treatment groups in Example 2.

[0031] Fig.14 It is a bar graph of the relative abundance percentage and ratio of Firmicutes and Bacteroidetes in the intestines of mice in different treatment groups in Example 2.

[0032] Fig.15 This is a heat map of the intestinal flora composition of mice in different treatment groups in Example 2. DETAILED DESCRIPTION

[0033] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0034] Example 1

[0035] (1) Isolation, identification and preservation of strains:

[0036] In November 2023, a new probiotic strain was isolated from the feces of healthy dogs in Beijing. Figure 1 , Figure 2 ) and 16S rRNA gene sequence (shown in sequence 1), the strain was identified as animal-associated Lactobacillus ( Ligilactobacillus animalis ), and named it animal-associated Lactobacillus LA-1.

[0037] On September 27, 2024, the animal-associated Lactobacillus LA-1 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No. 32098 and the classification name as animal-associated Lactobacillus Ligilactobacillus animalis .

[0038] (2) Artificial gastrointestinal fluid test:

[0039] After activating and culturing the animal combined Lactobacillus LA-1 for the second generation, the bacterial solution concentration was adjusted to OD 600 nm=0.7, added to artificial gastric juice (3 g pepsin dissolved in 1 L of pH=3, 0.2 M PBS buffer) at a 10% (v / v) inoculum, incubated at 37°C, and sampled and diluted to 10 at 0 h, 1.5 h, and 3 h. -6 , 10 -7 , 10 -8 Plate and count three replicates for each dilution. Transfer 1 mL of artificial gastric juice containing animal-associated Lactobacillus LA-1 into 100 mL of artificial intestinal juice (1 g of trypsin dissolved in 1 L of pH=8, 0.2 M PBS buffer), incubate at 37°C, and dilute to 10% at 0 h, 2 h, 4 h, and 8 h. -6 , 10 -7 , 10 -8 Plate and count 3 replicates for each dilution.

[0040] The results are as follows Figure 3 As shown, W1.5h and W3h represent the bacterial survival rates of the strains after being treated in artificial gastric juice for 1.5 h and 3 h, respectively, C0h, C2h, C4h, and C8h represent the bacterial survival rates of the strains after being treated in artificial intestinal juice for 0 h, 2 h, 4 h, and 8 h, respectively. The survival rates of Animal-associated Lactobacillus LA-1 in each time period were all above 100%, indicating that Animal-associated Lactobacillus LA-1 proliferated in artificial gastric juice and artificial intestinal juice, and therefore Animal-associated Lactobacillus LA-1 can fully play its role in gastrointestinal juice.

[0041] (3) Cholesterol-lowering trial:

[0042] The second generation culture of animal combined Lactobacillus LA-1 was inoculated into MRS-CHOL medium (1L MRS-CHOL medium containing 0.1 g cholesterol and 0.2 g ox bile salt) at 2% (v / v) and cultured in a constant temperature incubator at 37°C. After 48 h of cultivation, 0.50 mL of bacterial suspension was aspirated and mixed with 4.50 mL of anhydrous ethanol. After standing for 10 min, the mixture was centrifuged at 3000 r / min for 15 min. 0.50 mL of supernatant, 0.20 mL of o-phthalaldehyde solution, and 4.30 mL of mixed acid (concentrated sulfuric acid and glacial acetic acid = 1:1) were taken and shaken to prepare the experimental group. MRS-CHOL medium without inoculation of animal-associated Lactobacillus LA-1 was used instead of bacterial suspension and mixed with 4.50 mL of anhydrous ethanol. After standing for 10 min, the mixture was centrifuged at 3000 r / min for 15 min. 0.50 mL of supernatant, 0.20 mL of o-phthalaldehyde solution, and 4.30 mL of mixed acid were shaken to prepare the control group.

[0043] After the experimental and control groups were left to stand for 30 min, the 550 nmThe absorbance was measured at 40 °C and the cholesterol degradation rate was calculated. Cholesterol degradation rate = (CA) / C × 100%, where C is the OD of the control group. 550 nm The cholesterol content corresponding to the value, A is the OD of the experimental group 550 nm The cholesterol content corresponding to the value.

[0044] It was calculated that the cholesterol degradation rate of animal-associated Lactobacillus LA-1 was 50.03%.

[0045] (4) Bile salt hydrolase activity test:

[0046] The excretion of bile salts is the main way to eliminate cholesterol in the body and is also an important way of cholesterol metabolism.

[0047] Sterile filter papers were placed on MRS plates, sodium glycocholate plates (MRS plates + 0.35% (w / v) CaCl2 and 0.5% (w / v) sodium glycocholate hydrate) and sodium taurocholate plates (MRS plates + 0.35% (w / v) CaCl2 and 0.5% (w / v) sodium taurocholate hydrate), and then 20 μL of activated animal-associated Lactobacillus LA-1 culture liquid was inoculated on the sterile filter papers of the sodium glycocholate plates and sodium taurocholate plates, respectively, and anaerobically cultured at 37°C for 72 h.

[0048] Figure 4 The figure shows the filter paper photos of three plates after anaerobic culture for 72 hours. It can be found that white precipitate produced by the reaction of CaCl2 and free bile acid appeared around the filter paper inoculated with the animal-associated Lactobacillus LA-1 bacterial liquid, indicating that the animal-associated Lactobacillus LA-1 can decompose the bound bile salts into free bile salts when growing in the sodium taurocholate plate and the sodium glycocholate plate. Therefore, it can be concluded that the animal-associated Lactobacillus LA-1 has bile salt hydrolase activity, and also has sodium taurocholate and sodium glycocholate hydrolase activity.

[0049] (5) α-glucosidase inhibition test:

[0050] Inhibiting the activity of α-glucosidase can significantly delay the conversion of glucose into blood sugar, achieving the effect of lowering blood sugar.

[0051] The second generation of activated culture of animal-combined Lactobacillus LA-1 was inoculated in MRS medium at an inoculum of 2.0% (v / v) and cultured at 37°C for 24 h. 50 μL of the cultured bacterial solution was inoculated into 100 μL of 0.1 M phosphate buffer (pH=6.9) containing α-glucosidase and pre-incubated in a 96-well plate at 25°C for 10 min. After pre-incubation, 50 μL of 5 mM p-nitrophenyl-α-D-pyranoglucoside solution was added to each well. The reaction mixture was incubated at 25°C for 5 min, and the absorbance reading at 405 nm was recorded by a microplate reader. At the same time, 50 μL of 0.1 M phosphate buffer (pH=6.9) was used instead of the bacterial solution for the experiment as a control group.

[0052] It was calculated that the inhibition rate of animal combined Lactobacillus LA-1 on α-glucosidase was 54.54%.

[0053] (6) Fatty acid absorption test:

[0054] After activating and culturing the animal combined Lactobacillus LA-1 for the second generation, the bacterial solution concentration was adjusted to OD 600 nm =0.7, inoculated with an inoculum of 2.0% (v / v) in a medium containing 0.5% Brij 58 and 0.25 mmol / L sodium palmitate, and cultured at 37°C for 24 h. The concentration of fatty acids in the medium was measured using a free fatty acid determination kit, and the free fatty acid absorption rate of animal-associated Lactobacillus LA-1 was calculated to be 67.61%.

[0055] (7) Hydrophobicity test:

[0056] Surface hydrophobicity is an important factor affecting bacterial adhesion. Chloroform was used to evaluate cell surface hydrophobicity. 600 nm =0.7 (A0), 1 mL of bacterial solution was contacted with 0.25 mL of chloroform, the mixture was shaken for 2 min, and then allowed to stand at 37 °C for 1 h, and the absorbance of the aqueous phase was measured at 600 nm (A1).

[0057] According to the formula: surface hydrophobicity (%) = (1-A1 / A0) × 100, the surface hydrophobicity of animal-associated Lactobacillus LA-1 was calculated to be 75.85%.

[0058] (8) Hemolytic test:

[0059] Hemolysis is the rupture of red blood cells and the release of hemoglobin, which is called hemolysis of red blood cells, or hemolysis for short. It can be divided into three types: type A (α) hemolysis, type B (β) hemolysis, and type C (γ) hemolysis. Type A hemolysis: There is a grass-green hemolysis ring around the colony; type B hemolysis: There is a 2-4 mm transparent hemolysis ring around the colony; type C hemolysis: No hemolysis occurs.

[0060] The second generation of activated culture of animal combined Lactobacillus LA-1 was inoculated on Columbia blood agar plates and cultured at 37°C for 48 h. The changes around the colonies were observed, and Escherichia coli K88 was used as a positive control strain.

[0061] The results are as follows Figure 5 As shown, there is no hemolytic ring around the animal-associated Lactobacillus LA-1, which is type C (γ) hemolysis, that is, non-hemolysis, indicating that the animal-associated Lactobacillus LA-1 is safe.

[0062] (9) Antibiotic susceptibility test:

[0063] The sensitivity of animal Lactobacillus LA-1 to antibiotics was tested by the drug sensitivity paper agar diffusion method. The specific steps are as follows:

[0064] The second generation of activated culture of animal-associated Lactobacillus LA-1 was inoculated in MRS medium at an inoculum of 2.0% (v / v), cultured at 37℃ for 6 h, and 100 μL of bacterial solution was aspirated to plate. The drug-sensitive paper containing different antibiotics (tetracycline 30 μg / tablet, clindamycin 2 μg / tablet, gentamicin 10 μg / tablet, erythromycin 15 μg / tablet, chloramphenicol 30 μg / tablet, amcarcillin 10 μg / tablet) and positive drug-sensitive sheets were placed on the plate and cultured at 37℃ for 24 h to observe the sensitivity of animal-associated Lactobacillus LA-1 to drugs. The results are shown in Table 1. "S" indicates that there is an antibiotic sensitivity circle, and the bacteria are sensitive to antibiotics; "R" indicates that there is no antibiotic sensitivity circle, and the bacteria are not sensitive to antibiotics and have drug resistance. Animal-associated Lactobacillus LA-1 is sensitive to the common antibiotics tetracycline, clindamycin, gentamicin, erythromycin, chloramphenicol and amcarcillin tested, and is safe.

[0065] Table 1 Antibiotic sensitivity test results of animal combined lactobacillus LA-1

[0066]

[0067] Example 2

[0068] An obese mouse model was established by a high-fat diet to evaluate the obesity-alleviating and lipid-lowering effects of animal combined with Lactobacillus LA-1.

[0069] (1) Animal combined with Lactobacillus LA-1 culture:

[0070] After animal-associated Lactobacillus LA-1 was activated for two generations by MRS culture, it was inoculated in MRS medium at a 2% (v / v) inoculation volume and cultured at 37°C for 24 h. After the culture was completed, the bacteria were collected by centrifugation at 4°C and 5000 r / min for 10 min, washed twice with 0.9% saline, and the number of live bacteria of animal-associated Lactobacillus LA-1 was adjusted to 1×10 by plate live bacteria counting method. 10 CFU / mL.

[0071] (2) Experimental feeding and grouping:

[0072] Thirty-two healthy 7-week-old C57BL / 6J male mice were randomly divided into CT group, HFD group, positive control (PC group) and LA-1 group, with 8 mice in each group and 2 mice in each cage. After 7 days of adaptation, the CT group was fed with maintenance diet, and the HFD group, PC group and LA-1 group were fed with high-fat diet (D12492). The CT group and HFD group were gavaged with 200 μL of sterile 0.9% saline every day, the PC group was fed with the same volume of atorvastatin calcium tablets (30 mg / kg) every day, and the LA-1 group was gavaged with 200 μL of the animal combined lactobacillus LA-1 in step (1) every day for 16 consecutive weeks. The mice were kept at a temperature of 22±2℃, and they had free access to water and food during this period. At the end of the experiment, the mice were starved for 16 hours, weighed, and their weight changes were analyzed. After blood sampling, they were killed by cervical dislocation and their organs were separated.

[0073] (3) Weight changes:

[0074] The changes in mouse body weight Figure 6 As shown in the figure, compared with the CT group, the weight of the mice in the HFD group fed with a high-fat diet increased significantly, and the weight gain of the mice in the LA-1 group and the PC group slowed down significantly after treatment; compared with the PC group, the weight gain rate of the LA-1 mice slowed down further, especially from the 6th week to the 16th week, the weight gain rate of the mice in the LA-1 group was significantly lower than that in the PC group, indicating that the intervention effect of LA-1 is better than that of atorvastatin calcium tablets.

[0075] (4) Mouse organ index:

[0076] After autopsy, the liver, white fat, kidney and spleen were removed from the mice, rinsed with saline, dried with filter paper and weighed. The organ index was calculated according to the formula: organ index = (organ weight / body weight) × 100%.

[0077] The results are as follows Figure 7As shown, compared with the CT group, the liver and kidney weights of the HFD group increased slightly, the abdominal white fat weight increased significantly, and there was no significant difference in spleen weight among the groups; after animal combined Lactobacillus LA-1 intervention, the liver weight of mice decreased, and the white fat weight and kidney weight decreased significantly (P<0.05), and the degree of white fat reduction was greater than that of mice in the PC group. These results indicate that animal combined Lactobacillus LA-1 intervention can help alleviate fat and kidney organ hypertrophy, and the effect can replace atorvastatin calcium tablets.

[0078] (5) Liver lesions:

[0079] The liver tissues of mice in each treatment group were fixed with 4% tissue fixative, dehydrated, transparent, wax-impregnated, embedded, and sectioned, and then stained with Oil Red O (ORO) and HE. Finally, the changes in tissue morphology were observed under an optical microscope.

[0080] The results are as follows Figure 8 As shown, Oil Red O staining showed that compared with the CT group, the lipid enrichment in the HFD group was significantly increased, and the lipid enrichment in the LA-1 group mice was reduced, and the lipid enrichment was lower than that in the PC group; HE staining showed that compared with the CT group, the liver tissue of the HFD group mice had vesicular fatty degeneration, the fat content of the liver tissue increased, there were a large number of fat vacuoles, the cell contents flowed out, the cells were swollen, and the integrity of the cell wall was destroyed; after the intervention of the animal combined Lactobacillus LA-1 and PC group, the degree of fatty degeneration of the liver tissue of the mice was significantly alleviated, the fat vacuoles of the hepatocytes were smaller and fewer, and the morphology was similar to that of the CT group. At the same time, the results show that the LA-1 group can replace atorvastatin calcium tablets.

[0081] (6) Fatty lesions:

[0082] The diameter of fat vesicles can directly reflect the obesity degree of mice. The abdominal fat of mice in each treatment group was fixed with 4% tissue fixative, and pathological HE staining sections were made to observe tissue changes.

[0083] like Fig. 9 As shown in the figure, compared with the CT group, the diameter of the fat vesicles in the adipose tissue of the HFD group mice increased, and the diameter of the fat vesicles in the LA-1 group decreased, and the degree of reduction was better than that in the PC group. Compared with the PC group, the morphology of the effect in the LA-1 group was more similar to that in the CT group.

[0084] (7) Serum and liver index testing:

[0085] When the body consumes a high-fat diet for a long time, the body's fat decomposition will accelerate, inhibit lipid synthesis, produce excessive fatty acids, and increase serum triglyceride (TG) and total cholesterol (TC) levels. At the same time, when the body is obese, the low-density lipoprotein cholesterol (LDL-C) content increases. The liver is the largest detoxification organ and lipid metabolism center in the human body. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are important indicators for liver function diagnosis and can reflect the degree of liver abnormalities. Alanine aminotransferase and aspartate aminotransferase are mainly distributed in hepatocytes. When hepatocytes are necrotic, alanine aminotransferase and aspartate aminotransferase will be released into the blood circulation, causing serum enzymes to rise. The level of increase is positively correlated with the degree of liver tissue abnormalities. Lactate dehydrogenase (LDH) is an enzyme widely present in the cytoplasm of all tissue cells in the body. When tissues and organs are damaged, a large amount of lactate dehydrogenase will be released into the blood. At the same time, elevated lactate dehydrogenase is related to an increase in intestinal toxins, because intestinal toxins can damage the intestinal mucosal barrier function and increase the release of lactate dehydrogenase.

[0086] The levels of triglyceride, total cholesterol, low-density lipoprotein cholesterol, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase in the serum of mice in each treatment group were measured according to the kit instructions.

[0087] The results are as follows Fig.10 As shown in the figure, compared with the CT group, the levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, and alanine aminotransferase in the HFD group were significantly increased (P<0.05), and the levels of aspartate aminotransferase and lactate dehydrogenase were increased. Compared with the HFD group, the levels of triglyceride, alanine aminotransferase, and lactate dehydrogenase in the LA-1 group were significantly decreased (P<0.05), and the levels of total cholesterol, low-density lipoprotein cholesterol, and aspartate aminotransferase were decreased; compared with the PC group, the levels of alanine aminotransferase and lactate dehydrogenase in the LA-1 group were significantly decreased (P<0.05), and the levels of triglyceride, total cholesterol, low-density lipoprotein cholesterol, and aspartate aminotransferase were decreased. The results showed that animal combined with Lactobacillus LA-1 intervention has the effect of regulating serum lipids and protecting liver function, and its effect of reducing endotoxin is better than that of atorvastatin calcium tablets.

[0088] According to the instructions of the kit, triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, glucose, alanine aminotransferase, and aspartate aminotransferase in the liver of mice in each treatment group were measured.

[0089] The results are as follows Fig.11As shown, compared with the CT group, the levels of triglycerides and alanine aminotransferase in the mice in the HFD group were significantly increased (P<0.05); the content of very low-density lipoprotein cholesterol was significantly decreased (P<0.05), the high-density lipoprotein cholesterol was decreased, and the levels of total cholesterol, low-density lipoprotein cholesterol, glucose, and aspartate aminotransferase were increased. Compared with the HFD group, the levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and aspartate aminotransferase in the mice in the LA-1 group after intervention were significantly decreased (P<0.05); the levels of glucose and alanine aminotransferase were decreased; the levels of high-density lipoprotein cholesterol and very low-density lipoprotein cholesterol were increased. Compared with the PC group, the low-density lipoprotein cholesterol and alanine aminotransferase in the mice in the LA-1 group were significantly decreased (P<0.05); the total cholesterol, triglycerides, glucose, and aspartate aminotransferase were decreased, and the content of very low-density lipoprotein cholesterol was increased. These results indicate that the comprehensive effect of animal combined with Lactobacillus LA-1 in regulating liver lipids and protecting liver function is better than atorvastatin calcium tablets.

[0090] (9) Changes in fecal flora:

[0091] The PCoA analysis results of mice in each treatment group are shown in Fig.12 As shown in the figure, the HFD group was separated from the CT group, indicating that feeding a high-fat diet caused changes in the intestinal flora. Compared with the HFD group, there were differences between the LA-1 group and the PC group, indicating that animal-associated Lactobacillus LA-1 had a significant regulatory effect on the intestinal flora.

[0092] Analyze the intestinal flora structure, such as Fig.13 As shown, the stacked columns are the top 10 taxa at the phylum level in the intestinal tract of mice in each treatment group. Fig.14 As shown in Table 2 below, the ratio of Firmicutes / Bacteroidetes in the intestines of mice in the CT group was 37%, and the ratio of Firmicutes / Bacteroidetes in the intestines of mice in the HFD group was significantly increased to 280%. The increase in the ratio of Firmicutes / Bacteroidetes is a major feature of the intestinal flora structure of obesity and cardiovascular diseases; the ratio of Firmicutes / Bacteroidetes in the intestines of mice in the LA-1 group was significantly decreased to 189%, indicating that animal combined with Lactobacillus LA-1 can alleviate the disorder of intestinal flora structure caused by a high-fat diet by regulating the intestinal flora, and the effect is better than that of atorvastatin calcium tablets (198%).

[0093] Table 2 Relative abundance and ratio of Firmicutes and Bacteroidetes in the intestines of mice in each group

[0094]

[0095] At the genus level, the structure of the intestinal flora was analyzed, e.g. Fig.15As shown, the color gradient from blue to red indicates that the abundance of bacterial genera in different samples ranges from low to high, and the colors correspond to the legend. At the genus level, compared with the CT group, in the HFD group, the abundance of Bacteroides ( Bacteroides ), another genus Mycobacterium ( Alistipes Compared with the HFD group, the LA-1 group had an increase in the number of Bacteroides ( Bacteroides ), Faecalibacterium ( Faecalibaculum )、Desulfovibrio spp.( Desulfovibrio ) and Lachnospiraceae ( Lachnoclostridium ), and the intestinal flora structure was similar to that of the PC group. This indicates that animal-combined Lactobacillus LA-1 can regulate intestinal flora by increasing the relative abundance of bacteria that produce short-chain fatty acids, thereby reducing body weight and regulating lipid metabolism. At the same time, the effect of LA-1 group in regulating intestinal flora structure was similar to that of the PC group, and animal-combined Lactobacillus LA-1 can replace atorvastatin calcium tablets.

[0096] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A strain of animal-associated Lactobacillus ( Ligilactobacillus animalis ) LA-1, characterized in that It was deposited in the General Microbiology Center of China Microbiological Culture Collection on September 27, 2024. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.32098.

2. A microbial preparation comprising the animal-associated Lactobacillus LA-1 as claimed in claim 1.

3. The microbial preparation according to claim 2, characterized in that The microbial preparation is a liquid preparation or a solid preparation.

4. The microbial preparation according to claim 3, characterized in that The content of animal combined lactobacillus LA-1 in the liquid preparation is 1.0×10 7 ~1.0×10 10 CFU / mL.

5. The microbial preparation according to claim 3, characterized in that The content of animal combined lactobacillus LA-1 in the solid preparation is 1.0×10 7 ~1.0×10 10 CFU / g.

6. Use of the animal-associated Lactobacillus LA-1 as claimed in claim 1 or the microbial preparation as claimed in any one of claims 2 to 5 in the preparation of medicines, characterized in that: The drug can be used to (1) prevent, alleviate or treat obesity; (2) prevent, alleviate or treat metabolic syndrome; (3) prevent, alleviate or treat hyperlipidemia.

7. The use according to claim 6, characterized in that Animal-associated Lactobacillus LA-1 has the following functions: (a) Reduce triglyceride content in the liver and serum; (b) Reduce total cholesterol content in the liver, reduce low-density lipoprotein cholesterol, and increase very low-density lipoprotein cholesterol; (c) Reduce liver and serum alanine aminotransferase and aspartate aminotransferase levels, enhance liver function, and protect the liver; (d) Reduce serum lactate dehydrogenase; (e) Improve liver and fat pathological characteristics; (f) Regulate intestinal flora.

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