Murine combined lactobacillus and application thereof in treating non-alcoholic fatty liver disease induced by choline tryptophan deficiency

Abstract

The invention discloses a strain of lactobacillus murinus and application thereof in treating non-alcoholic fatty liver disease induced by choline tryptophan deficiency, through verification of the strain of lactobacillus murinus with the preservation number of CCTCC M 20242901, the strain of lactobacillus murinus has the effects of reducing tissue fat deposition on the non-alcoholic fatty liver disease induced by choline tryptophan deficiency, remarkably reducing the levels of ALT and AST in serum, and improving the treatment effect on the non-alcoholic fatty liver disease induced by choline tryptophan deficiency. The content of triglyceride (TG) in liver tissue is reduced, the compound can be used for preventing or treating the non-alcoholic fatty liver disease induced by choline tryptophan deficiency, and a new choice is provided for the medicine for preventing and treating the non-alcoholic fatty liver disease induced by choline tryptophan deficiency.

Description

Technical Field

[0001] The present invention relates to the technical field of drugs for the treatment of non - alcoholic fatty liver disease, and specifically to a strain of Ligilactobacillus murinus and its application in the treatment of non - alcoholic fatty liver disease induced by choline - tryptophan deficiency. Background Art

[0002] Choline is an essential nutrient for the human body, belonging to the B - group vitamins. It participates in the synthesis of phospholipids and cholesterol and is crucial for maintaining the normal function of the liver. Choline can promote the excretion of fat from the liver in the form of phospholipids; it can improve the utilization of fatty acids in the liver and prevent the excessive accumulation of fat in the liver. Research shows that the lack of choline will affect the liver's handling of fat, causing fat to accumulate in the liver, thus triggering non - alcoholic fatty liver disease (NAFLD). Supplementing choline can help improve the lipid metabolism of the liver and reduce the accumulation of liver fat.

[0003] Choline not only plays an important role in liver health but is also closely related to the balance of the gut microbiota. Sufficient intake of choline in the diet can promote the growth of beneficial bacteria, thus contributing to maintaining gut health and immune function. Gut bacteria can metabolize choline into trimethylamine (TMA), which is then converted by the liver into trimethylamine N - oxide (TMAO). TMAO is associated with an increased risk of cardiovascular disease. In addition, choline intake may affect the diversity and abundance of gut bacteria, and thus have an impact on the metabolism and health of the host. Improving the metabolism of choline by modulating bacteria will be an important means for the treatment of NAFLD. Summary of the Invention

[0004] The present invention provides a strain of Ligilactobacillus murinus and its application in the treatment of non - alcoholic fatty liver disease induced by choline - tryptophan deficiency. By isolating a strain of Ligilactobacillus murinus that can improve choline metabolism from the feces of intestinal epithelial cell - specific Fut2 knockout (Fut2 △IEC ) mice, and verifying its efficacy in treating non - alcoholic fatty liver disease induced by choline - tryptophan deficiency (MCD), it provides a new option for the prevention and treatment of drugs for non - alcoholic fatty liver disease induced by choline - tryptophan deficiency.

[0005] In view of this, the solution of the present invention is as follows:

[0006] In the first aspect of the present invention, a strain of Ligilactobacillus murinus is proposed, which is preserved in the China Center for Type Culture Collection with the preservation number CCTCC M20242901.

[0007] The second aspect of the present invention lies in providing a drug for preventing or treating a disease, wherein the drug contains the Lactobacillus rhamnosus strain or its bacterial solution described in the first aspect, and the disease is non-alcoholic fatty liver induced by choline tryptophan deficiency.

[0008] Further, the unit dose of the drug contains 1×10 9 、2×10 9 、3×10 9 、4×10 9 、5×10 9 、6×10 9 、7×10 9 、8×10 9 or 9×10 9 CFU or more of Lactobacillus rhamnosus.

[0009] Further, the drug is in the form of a freeze-dried powder or tablets.

[0010] Further, the drug contains pharmaceutically acceptable adjuvants.

[0011] Preferably, the adjuvants include carriers, and / or diluents, and / or excipients.

[0012] The third aspect of the present invention lies in providing the application of the Lactobacillus rhamnosus strain or its bacterial solution described in the first aspect, and the application is for preparing a drug for preventing or treating a disease, wherein the disease is non-alcoholic fatty liver induced by choline tryptophan deficiency.

[0013] In the above application, the drug is for oral administration.

[0014] In the above application, the drug has at least one of the following uses:

[0015] a) Reducing fat deposition in liver tissue;

[0016] b) Reducing hepatocyte damage;

[0017] c) Lowering the levels of serum ALT and AST;

[0018] d) Lowering the content of triglyceride in liver tissue.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] A strain of Ligilactobacillus murinus with the preservation number of CCTCC M 20242901 provided by the present invention has been verified to have the effects of reducing tissue fat deposition, significantly reducing the levels of serum ALT and AST, and reducing the content of triglyceride (TG) in liver tissue for non-alcoholic fatty liver induced by choline tryptophan deficiency. It can be used as a drug for preventing or treating non-alcoholic fatty liver induced by choline tryptophan deficiency, providing a new option for the prevention and treatment drugs of non-alcoholic fatty liver induced by choline tryptophan deficiency. Brief Description of the Drawings

[0021] Figure 1 It is the experimental results after PBS and Ligilactobacillus murinus were respectively intragastrically administered to the control and MCD-fed mice in Example 2 of the present invention; wherein Figure A is the HE staining of liver tissue; Figure B is the change of serum ALT and AST levels, P < 0.05; Figure C is the change of triglyceride (TG) content in liver tissue, P < 0.05. Detailed Description of the Invention

[0022] The technical solutions of the present invention will be clearly and completely described below in conjunction with the preferred embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0023] Example 1

[0024] Our previous research found that Fut2 △IEC mice can resist MCD-induced non-alcoholic fatty liver. Therefore, we collected the feces of Fut2 △IEC mice and carried out anaerobic culture using Schaedler broth medium to screen out a potential probiotic strain that rarely consumes choline in the medium. The 16S rDNA fragment of this strain was amplified using the 27F / 1492R universal primer, and the target gene sequence was obtained by sequencing, as shown in SEQ ID NO: 1, specifically:

[0025]

[0026] The sequencing sequence was aligned with the nt database of NCBI, and the NCBI taxonomy database was retrieved to determine that the strain was Ligilactobacillus murinus CH2022. This strain was deposited at the China Center for Type Culture Collection (CCTCC) on December 25, 2025, with the deposit number CCTCC M20242901.

[0027] Example 2

[0028] The activated Ligilactobacillus murinus bacterial solution after 2 generations was inoculated into Schaedler broth medium at an inoculation amount of 1%, cultured anaerobically and statically at 37 °C for 36 h, then counted, and centrifuged at 3000 rpm / min for 15 min to retain the precipitate. It was diluted to a concentration of 2×10 9 with a cryopreservation solution prepared with 30% glycerol bacterial solution, and stored at -89 °C at 200 μL per tube. The bacterial solution was prepared once a week and rewarmed in a 37 °C water bath before gavage.

[0029] Ten-week-old male C57BL / 6 mice were fed normal diet and MCD diet respectively, and were gavaged with Ligilactobacillus murinus and an equal amount of PBS respectively. They were divided into a control diet + PBS group, an MCD + PBS group, a control diet + Ligilactobacillus murinus group, and an MCD + Ligilactobacillus murinus group. There were 8 mice in each group. The MCD diet was fed for 30 days, and PBS (200 μL) or Ligilactobacillus murinus (200 μL) was gavaged every 3 days. The hepatic tissue fat deposition and stem cell damage changes were detected by HE staining of the liver tissue, and the serum ALT, AST levels, and the content of triglyceride (TG) in the liver tissue were detected. The results were as follows Figure 1 shown. Compared with the MCD + PBS group, the hepatic tissue fat deposition and hepatocyte damage in the MCD + Ligilactobacillus murinus group were significantly reduced (Figure A, P < 0.05); compared with the MCD + PBS group, the serum ALT and AST levels in the MCD + Ligilactobacillus murinus group were significantly decreased (Figure B, P < 0.05); compared with the MCD + PBS group, the content of triglyceride (TG) in the liver tissue of the MCD + Ligilactobacillus murinus group decreased (Figure C, P < 0.05).

[0030] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A strain of Lactobacillus murinus, which is deposited in China Center for Type Culture Collection with a deposit number of CCTCC M 20242901.

2. A drug for preventing or treating a disease, characterized in that: The medicine comprises the mouse Lactobacillus strain or its bacterial liquid according to claim 1, and the disease is non-alcoholic fatty liver induced by choline tryptophan deficiency.

3. The drug according to claim 2, characterized in that A unit dose of the drug contains 1×10 9 , 2×10 9 , 3×10 9 , 4×10 9 , 5×10 9 , 6×10 9 ,7×10 9 , 8×10 9 or 9×10 9 CFU or above of Lactobacillus murineus.

4. The drug according to claim 2, characterized in that The drug is in the form of freeze-dried powder or tablet.

5. The drug according to claim 2, characterized in that The medicament contains a pharmaceutically acceptable adjuvant.

6. The drug according to claim 5, characterized in that The auxiliary agent includes a carrier, and / or a diluent, and / or an excipient.

7. The use of Lactobacillus murineus according to claim 1, characterized in that: The application is to prepare medicine for preventing or treating diseases, wherein the disease is non-alcoholic fatty liver induced by choline tryptophan deficiency.

8. The use according to claim 7, characterized in that: The drug is for oral administration.

9. The use according to claim 7, characterized in that: The drug includes at least one of the following uses: a) Reduce fat deposition in liver tissue; b) Reduce liver cell damage; c) Reduce serum ALT and AST levels; d) Reduce the triglyceride content in liver tissue.

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