A fermented mucus lactobacillus YDJ-6 and its application

Abstract

The present invention provides a fermented mucus lactobacillus YDJ-6 and an application thereof, wherein the fermented mucus lactobacillus YDJ-6 is deposited in the General Microbiological Center of China Microbiological Culture Collection Committee, with a deposit number of CGMCC No.28036, a deposit time of July 27, 2023, and an address located at the Institute of Microbiology, Chinese Academy of Sciences. The fermented mucus lactobacillus described in the present invention not only has good stability under normal temperature and freezing conditions, maintains stable activity, has a long shelf life, has high adsorption to intestinal cells, has a high survival rate in a simulated gastric acid bile salt environment, has good application safety, and can reduce the uric acid concentration in the blood, regulate blood sugar and blood lipids, improve hyperlipidemia, liver damage, and kidney damage, and has certain antioxidant and anti-inflammatory functions.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to a fermented mucus lactobacillus YDJ-6 and an application thereof. Background Art

[0002] With the acceleration of modern life and changes in eating habits, hyperuricemia, liver disease, metabolic syndrome and the accompanying oxidative stress and inflammatory response have become important challenges in the field of global public health. Hyperuricemia is not only directly related to gout, but also significantly increases the risk of chronic diseases such as cardiovascular disease, diabetes and non-alcoholic fatty liver disease. In addition, liver fat accumulation, liver function damage and systemic metabolic abnormalities caused by a high-fat diet are also the focus of current medical attention.

[0003] In the process of exploring these disease prevention and treatment strategies, probiotics have gradually gained favor among researchers and consumers due to their unique physiological functions and safety. In particular, Lactobacillus fermentum, as an important member of the probiotic family, has been proven to have a variety of health benefits, including regulating the balance of intestinal flora, enhancing immunity, and promoting nutrient absorption. However, most probiotic products on the market currently have problems such as single function, unstable activity, short shelf life, and unverified application safety, which makes it difficult to meet the growing health needs.

[0004] In view of the above technical background and the deficiencies of the prior art, the present invention aims to provide a fermented lactobacillus with multiple functions such as lowering uric acid, protecting the liver, regulating blood sugar and blood lipids, anti-oxidation and anti-inflammatory, which can not only effectively reduce blood uric acid levels, relieve hyperuricemia and its complications, but also significantly relieve liver damage and improve renal damage caused by hyperuricemia. At the same time, the fermented lactobacillus has excellent stability, can maintain stable activity under normal temperature and freezing conditions, and has a long shelf life. Summary of the invention

[0005] The purpose of the present invention is to provide a fermented mucus lactobacillus YDJ-6 and an application thereof. The fermented mucus lactobacillus not only has good stability under normal temperature and freezing conditions, maintains stable activity, has a long shelf life, has high adsorption to intestinal cells, has a high survival rate in a simulated gastric acid and bile salt environment, has good application safety, but also can reduce the uric acid concentration in the blood, regulate blood sugar and blood lipids, improve hyperlipidemia, liver damage, and kidney damage, and has certain antioxidant and anti-inflammatory functions.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The fermented Lactobacillus mucilaginosus ( limosilactobacillus fermentum)YDJ-6, deposited in the General Microbiology Center of China Microbiological Culture Collection, with the deposit number: CGMCC No.28036, and the deposit date is July 27, 2023. The deposit address is located at Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0008] The fermented mucus lactobacillus YDJ-6 is obtained from a healthy intestine.

[0009] The fermented mucus lactobacillus YDJ-6 has the ability to lower blood lipids.

[0010] The fermented mucus lactobacillus YDJ-6 has the ability to reduce uric acid.

[0011] The fermented Lactobacillus mucilaginosus YDJ-6 has the ability to improve liver damage.

[0012] The fermented Lactobacillus mucilaginosus YDJ-6 has the ability to improve kidney damage.

[0013] The fermented mucus lactobacillus YDJ-6 has the ability of resisting oxidation.

[0014] The fermented Lactobacillus mucilaginosus YDJ-6 has anti-inflammatory ability.

[0015] The fermented mucus lactobacillus YDJ-6 has good stability under normal temperature and freezing conditions, and its activity is maintained stably.

[0016] The fermented mucus lactobacillus YDJ-6 has high adsorption to intestinal cells and high survival rate in a simulated gastric acid and bile salt environment.

[0017] The fermented mucus lactobacillus YDJ-6 can be used in preparing food and can be used as a food additive and added to various foods, such as yogurt, lactic acid bacteria beverages, juices, bread, biscuits, and the like.

[0018] The fermented mucus lactobacillus YDJ-6 can be used to prepare medicines or health products, and can be prepared into dosage forms such as oral liquid, powder, gel, suppository, hard capsule, soft capsule and the like.

[0019] The fermented mucus lactobacillus YDJ-6 can be used in the preparation of a lipid-lowering composition, a uric acid-lowering composition, a liver injury-improving composition, a kidney injury-improving composition, and an antioxidant or anti-inflammatory composition.

[0020] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0021] 1. The fermented mucus lactobacillus YDJ-6 prepared by the present invention has good stability at room temperature and freezing conditions, maintains stable activity, and has a long shelf life; the fermented mucus lactobacillus YDJ-6 has high adsorption to intestinal cells, has a high survival rate in a simulated gastric acid and bile salt environment, and has good application safety.

[0022] 2. The fermented mucus lactobacillus YDJ-6 prepared by the present invention can not only reduce the uric acid concentration in the blood, regulate blood sugar and blood lipids, but also improve hyperlipidemia, liver damage and kidney damage in mice, and at the same time has certain antioxidant and anti-inflammatory functions.

[0023] 3. The present invention reduces hyperlipidemia in mice by reducing triglycerides and low-density lipoprotein cholesterol in mouse serum.

[0024] 4. The present invention improves liver damage in mice by reducing alanine aminotransferase, aspartate aminotransferase and malondialdehyde in mouse serum, and has a certain effect of reducing lipid peroxidation and protecting the liver.

[0025] 5. The present invention significantly reduces the levels of uric acid and creatinine in the serum of mice, alleviates kidney damage, reduces the swelling of renal tubular epithelial cells in mice, reduces the infiltration of renal inflammatory cells, improves the thickening of the glomerular basement membrane, mesangial cell proliferation, and increased extracellular matrix in mice, and improves renal function damage caused by high uric acid.

[0026] 6. The fermented mucus lactobacillus YDJ-6 described in the present invention is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.28036 and the deposit date July 27, 2023, at the Institute of Microbiology, Chinese Academy of Sciences. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art.

[0028] Figure 1 It is the survival rate of the fermented mucus lactobacillus YDJ-6 described in the present invention in a simulated gastric acid and bile salt environment.

[0029] Figure 2 These are photos and data of the adsorption of the fermented mucus lactobacillus YDJ-6 described in the present invention on intestinal cells.

[0030] Figure 3 This is the test data of reducing the concentration of uric acid in blood by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention.

[0031] Figure 4 This is the test data of regulating blood sugar and blood lipids by the fermented mucus Lactobacillus YDJ-6 described in the present invention.

[0032] Figure 5 These are the stability test data of the fermented mucus Lactobacillus YDJ-6 described in the present invention.

[0033] Figure 6 These are the data related to the improvement of hyperlipidemia in mice by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention.

[0034] Figure 7 This is a photo of HE staining of mouse liver by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention.

[0035] Figure 8 These are data related to the improvement of mouse liver damage by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention.

[0036] Fig. 9 This is a photo of HE staining of mouse kidney by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention.

[0037] Fig.10 This is a photo of PAS staining of mouse kidney by the fermented mucus lactobacillus YDJ-6 described in the present invention.

[0038] Fig.11 These are the data related to the improvement of mouse kidney damage by the fermented Lactobacillus mucilaginosus YDJ-6 described in the present invention. DETAILED DESCRIPTION

[0039] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] Example 1: Survival rate test of fermented mucus lactobacillus YDJ-6 in a simulated gastric acid and bile salt environment

[0041] After the fermented Lactobacillus mucosa YDJ-6 was placed in a simulated gastric acid bile salt environment (pH 3.5, bile concentration 0.3%) for 7 hours, its viable bacteria count was tested. The results are shown in Figure 1 .

[0042] from Figure 1 It can be seen that the fermented mucus Lactobacillus YDJ-6 still has a high number of viable bacteria and a high survival rate of viable bacteria after 7 hours in a simulated gastric acid and bile salt environment.

[0043] Example 2: Adsorption test of fermented Lactobacillus mucilaginosus YDJ-6 on intestinal cells

[0044] Grasp the coverslip with tweezers, blanch both sides, and place it in a 6-well plate for later use; cut the Caco-2 cells and adjust the cells, then transfer them to the 6-well plate for culture, and wait until the plate is full (referring to a full monolayer) before testing; replace the minimum essential culture medium (containing 10% fetal bovine serum) again 1 hour before the test; during the test, remove the culture medium, wash twice with PBS buffer, and add 1.5mL of bacterial solution (1×10 9 CFU / mL) and 1.5 mL of minimum essential medium (containing 10% fetal bovine serum) were mixed and cultured in a constant temperature incubator at 37°C and 5% CO 2 , 1~4h; after the incubation, wash twice with sterile PBS buffer, add methanol to fix the cells, remove the cover glass, wait until it is dry, and then dry the liquid on the fire; after the cover glass is gram-stained, count the number of bacteria under a microscope. The test results show the average of three repeated tests.

[0045] from Figure 2 It can be seen that under the 1500-fold microscopic field of view, the amount of bacterial adsorption of fermented mucus lactobacillus YDJ-6 on intestinal cells Caco-2 cells is greater than 100, indicating that fermented mucus lactobacillus YDJ-6 has a strong adsorption ability on intestinal cells.

[0046] Example 3: Test on reducing blood uric acid concentration by fermentation of Lactobacillus mucilaginosus YDJ-6

[0047] 53 patients with hyperuricemia, 9 in the placebo group, 9 in the YDJ-6 group, 10 in the TSR332 group, and 25 in the YDJ-6+TSR332 group, took the medicine three times a day, with a daily intake of 2×10 10 CFU, of which TSR332 is Lactobacillus reuteri TSR332 produced by Hangzhou Target Food Technology Co., Ltd. The trial period was 60 days. Blood was drawn on the 0th, 30th and 60th days to analyze the uric acid content and calculate the change rate of uric acid. The results are shown in Figure 3 .

[0048] from Figure 3 It can be seen that after 60 days of consumption, the concentration of uric acid in the blood can be significantly reduced.

[0049] Example 4: Fermentation of Lactobacillus mucilaginosus YDJ-6 to regulate blood sugar and blood lipids

[0050] 25 patients with hyperuricemia were compared before and after taking YDJ-6. They took it 3 times a day, with a daily intake of 2×10 10CFU, of which TSR332 is Lactobacillus reuteri TSR332 produced by Hangzhou Target Food Technology Co., Ltd. The trial period was 60 days, and blood was drawn on the 0th, 30th and 60th days to test the uric acid, blood sugar and cholesterol content, and calculate their change rates. Figure 4 .

[0051] from Figure 4 It can be seen that consuming YDJ-6 for 60 days can significantly reduce the levels of uric acid, blood sugar and cholesterol in the blood.

[0052] Example 5: Stability test of fermented Lactobacillus mucilaginosus YDJ-6

[0053] The fermented Lactobacillus mucilaginosus YDJ-6 was stored at 25℃ and -25℃ for 2 years, and the number of viable bacteria was tested every month. The results are shown in Figure 5 .

[0054] from Figure 5 It can be seen that the fermented mucus Lactobacillus YDJ-6 has good stability at room temperature and low temperature, and its activity remains stable during long-term storage.

[0055] Example 6: In vivo testing of fermented Lactobacillus mucilaginosus YDJ-6 in mice

[0056] Experimental methods: Mice were divided into 3 groups: normal, model, and fermented Lactobacillus YDJ-6, with 10 mice in each group. Except for the normal group, which was given a control diet (D09100304, Research Diets, USA), the other 2 groups of mice were fed with a high-fat, high-fructose, high-cholesterol diet (D09100310, Research Diets, USA) supplemented with yeast powder and adenine for 90 days to induce a hyperlipidemia and hyperuricemia model.

[0057] After the model was successfully established, the mice were gavaged with drugs every day, with a dose of 1.2×10 9 CFU count. The normal group and the model group were gavaged with pure water every day as a blank control. After 28 days of administration, the blood was collected from the inner canthus of the eye on an empty stomach, placed at room temperature for 2 h, centrifuged at 4 ° C and 4500 r / min for 10 min, and serum was collected. The uric acid (UA), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), blood glucose, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and other indicators in the serum of mice were determined using a veterinary automatic biochemical analyzer.

[0058] The mice were killed, and the left lobe of the liver and the left kidney (longitudinal section) were taken and immersed in formalin for preparation of pathological sections and stained with hematoxylin-eosin (HE). Renal pathology was also stained with periodic acid-Schiff (PAS). About 0.1 g of liver and kidney tissues were taken and homogenized with about 0.9 mL of normal saline for determination of malondialdehyde (MDA). The experimental results are shown in Figure 6~Figure 11 .

[0059] from Figure 6 It can be seen that YDJ-6 significantly reduces triglycerides and low-density lipoprotein cholesterol in mouse serum, indicating that YDJ-6 has a significant lipid-lowering effect.

[0060] from Figure 7 It can be seen that compared with the normal group, a large number of lipid droplets appeared in the liver of the model group, while fermented Lactobacillus YDJ-6 significantly reduced the deposition of lipid droplets in the liver. Figure 8 It can be seen that the levels of alanine aminotransferase and aspartate aminotransferase in YDJ-6 were significantly decreased compared with the model group, showing a significant effect in improving liver damage. The malondialdehyde level in the model group was significantly increased compared with the normal group, while YDJ-6 significantly reduced the malondialdehyde level in the liver, indicating that the liver protection effect of YDJ-6 is closely related to reducing lipid peroxidation.

[0061] from Fig. 9 It can be seen that the renal cells of mice in the normal control group were closely arranged, the renal tubules were clearly outlined, the cells were not swollen, and there were no desquamated cells or granular substances in the tubular lumen; the renal tubular epithelial cells of mice in the model group were severely swollen or granular vacuolar degeneration, the tubular lumen was significantly dilated, granules or cell casts were visible in the tubular lumen, and a large number of inflammatory cells were infiltrated; the renal tubular epithelial cells of mice in the fermentation Lactobacillus YDJ-6 group were slightly to moderately swollen, and the inflammatory cell infiltration was reduced. Fig.10 It can be seen that compared with the kidneys of the normal group of mice, the glomerular basement membrane of the model group of mice was thickened, the mesangial cells proliferated, and the extracellular matrix increased. The administration of fermented lactobacillus YDJ-6 significantly improved the above pathological conditions. Fig.11 It can be seen that compared with the normal control group, the serum uric acid and creatinine levels in the model group increased significantly, and the administration of fermented Lactobacillus YDJ-6 significantly reduced the uric acid and creatinine levels. Therefore, fermented Lactobacillus YDJ-6 can significantly reduce the levels of uric acid and creatinine and alleviate kidney damage.

[0062] The fermented mucus lactobacillus YDJ-6 prepared by the method described in the present application not only has good stability at room temperature and frozen conditions, maintains stable activity, has a long shelf life, has high adsorption to intestinal cells, has a high survival rate in a simulated gastric acid and bile salt environment, has good application safety, but also can reduce the uric acid concentration in the blood, regulate blood sugar and blood lipids, improve hyperlipidemia, liver damage, and kidney damage, and has certain antioxidant and anti-inflammatory functions.

[0063] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A fermented mucus lactobacillus ( limosilactobacillus fermentum )YDJ-6, characterized in that: The fermented mucus lactobacillus YDJ-6 is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.28036 and the deposit time being July 27, 2023.

2. The fermented mucus lactobacillus YDJ-6 according to claim 1, characterized in that The fermented mucus lactobacillus YDJ-6 is obtained from the intestinal tract.

3. Use of the fermented mucus Lactobacillus YDJ-6 according to any one of claims 1 or 2 in preparing food.

4. Use of the fermented mucus lactobacillus YDJ-6 according to any one of claims 1 or 2 in the preparation of health products.

5. The use according to claim 4, characterized in that: The dosage form of health care products is oral liquid, powder, gel, suppository, hard capsule or soft capsule.

6. Use of the fermented mucinous Lactobacillus YDJ-6 according to any one of claims 1 or 2 in preparing a lipid-lowering composition.

7. Use of the fermented mucus Lactobacillus YDJ-6 according to any one of claims 1 or 2 in the preparation of a uric acid-lowering composition.

8. A food prepared by fermenting Lactobacillus mucilaginosus YDJ-6 according to any one of claims 1 or 2.

9. A health product or medicine prepared by fermenting Lactobacillus mucilaginosus YDJ-6 according to any one of claims 1 or 2.

10. The health product or medicine according to claim 9, characterized in that: The dosage form of health products or medicines is oral liquid, powder, gel, suppository, hard capsule or soft capsule.

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