Lactobacillus murinus and application thereof in treatment of non-alcoholic fatty liver induced by choline and tryptophan deficiency

Abstract

The application discloses a Ligilactobacillus murinus and application thereof in treatment of choline and tryptophan deficiency-induced non-alcoholic fatty liver disease, and verifies a Ligilactobacillus murinus with a preservation number of CCTCC M 20242901. The Ligilactobacillus murinus can reduce tissue fat deposition, significantly reduce serum ALT and AST levels, and reduce liver tissue triglyceride (TG) content, and can be used for preventing or treating choline and tryptophan deficiency-induced non-alcoholic fatty liver disease, and provides a new choice for prevention and treatment drugs of choline and tryptophan deficiency-induced non-alcoholic fatty liver disease.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology for the treatment of non-alcoholic fatty liver disease, specifically to a strain of *Lactobacillus murineis* and its application in the treatment of choline tryptophan deficiency-induced non-alcoholic fatty liver disease. Background Technology

[0002] Choline is an essential nutrient for the human body, belonging to the B vitamins. It participates in the synthesis of phospholipids and cholesterol, and is crucial for maintaining normal liver function. Choline promotes 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 excessive fat accumulation. Studies have shown that choline deficiency affects the liver's processing of fat, leading to fat accumulation in the liver and thus causing non-alcoholic fatty liver disease (NAFLD). Choline supplementation can help improve lipid metabolism in the liver and reduce fat accumulation.

[0003] Choline plays a vital role not only in liver health but is also closely related to the balance of the gut microbiota. Adequate dietary choline intake can promote the growth of beneficial bacteria, thus helping to maintain gut health and immune function. Gut bacteria metabolize choline into trimethylamine (TMA), which is then converted into trimethylamine oxide (TMAO) by the liver. TMAO is associated with an increased risk of cardiovascular disease. Furthermore, choline intake may affect the diversity and abundance of gut bacteria, thereby influencing host metabolism and health. Improving choline metabolism through bacterial regulation will be an important approach in the treatment of NAFLD. Summary of the Invention

[0004] This invention provides a strain of *Lactobacillus assemblica* and its application in treating choline-tryptophan deficiency-induced non-alcoholic fatty liver disease, achieved by specifically knocking out Fut2 from intestinal epithelial cells (Fut2...). △IEC A strain of *Lactobacillus assemblica* that can improve choline metabolism was isolated from mouse feces, and its efficacy in treating non-alcoholic fatty liver disease induced by choline tryptophan deficiency (MCD) was verified, providing a new option for the prevention and treatment of 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] The first aspect of the present invention is to provide a strain of *Ligilactobacillus murinus*, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC M20242901.

[0007] A second aspect of the invention is to provide a medicament for the prevention or treatment of a disease comprising the *Lactobacillus murineis* strain or its culture as described in the first aspect, wherein the disease is choline tryptophan deficiency-induced non-alcoholic fatty liver disease.

[0008] Furthermore, 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 Mouse-associated lactobacillus with a CFU or higher.

[0009] Furthermore, the drug is in the form of lyophilized powder or tablets.

[0010] Furthermore, the drug contains pharmaceutically acceptable excipients.

[0011] Preferably, the excipients include a carrier, and / or a diluent, and / or a formaldehyde excipient.

[0012] A third aspect of the invention is the application of the strain of *Lactobacillus murineis* or its culture as described in the first aspect, said application being the preparation of a drug for the prevention or treatment of a disease, said disease being choline tryptophan deficiency-induced non-alcoholic fatty liver disease.

[0013] In the above applications, the drug is used orally.

[0014] In the above applications, the drug includes at least one of the following uses:

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

[0016] b) Reduce hepatocellular damage;

[0017] c) Reduce serum ALT and AST levels;

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

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

[0020] The present invention provides a strain of *Ligilactobacillus murinus* with accession number CCTCC M 20242901, which has been verified to reduce tissue fat deposition, significantly lower serum ALT and AST levels, and reduce liver tissue triglyceride (TG) content in choline tryptophan deficiency-induced non-alcoholic fatty liver disease. It can be used as a drug for the prevention or treatment of choline tryptophan deficiency-induced non-alcoholic fatty liver disease, providing a new option for the prevention and treatment of choline tryptophan deficiency-induced non-alcoholic fatty liver disease. Attached Figure Description

[0021] Figure 1 The figures show the experimental results of mice fed with control and MCD diets in Example 2 of this invention after being administered PBS and mouse combined with Lactobacillus by gavage, respectively; Figure A shows the HE staining of liver tissue; Figure B shows the changes in serum ALT and AST levels, P < 0.05; Figure C shows the changes in triglyceride (TG) content in liver tissue, P < 0.05. Detailed Implementation

[0022] The technical solution of the present invention will now be clearly and completely described in conjunction with preferred embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] Example 1

[0024] Our preliminary research found Fut2 △IEC Mice are resistant to MCD-induced non-alcoholic fatty liver disease, therefore we collected Fut2 mice. △IEC Mouse fecal matter was anaerobically cultured in Schaedler broth to screen for a potential probiotic strain that minimally consumes choline in the culture medium. The 16S rDNA fragment of this strain was amplified using universal primers 27F / 1492R, and the target gene sequence was obtained by sequencing, as shown in SEQ ID NO: 1.

[0025]

[0026] The sequencing sequence was compared with the NCBI nt database and the NCBI taxonomy database was searched to identify the strain as *Ligilactobacillus murinus* CH2022. This strain was deposited at the China Center for Type Culture Collection (CCTCC) on December 25, 2025, with accession number CCTCC M20242901.

[0027] Example 2

[0028] The activated second-generation *Lactobacillus murineis* bacterial culture was inoculated into Schaedler broth at a 1% inoculum size and anaerobically cultured at 37°C for 36 hours. After counting, the bacterial cells were centrifuged at 3000 rpm for 15 minutes, and the precipitate was retained. The precipitate was diluted with a cryopreservation solution prepared with 30% glycerol to a concentration of 2 × 10⁻⁶ cells / mL. 9 Concentration: 200 μL per tube, frozen at -89°C. Prepare bacterial suspension weekly, and warm the suspension in a 37°C water bath before gavage.

[0029] Ten-week-old male C57BL / 6 mice were fed either a standard diet or an MCD diet, and were then administered either mouse-derived Lactobacillus simulans or an equal volume of PBS via gavage. The mice were divided into four groups: control diet + PBS, MCD + PBS, control diet + mouse-derived Lactobacillus simulans, and MCD + mouse-derived Lactobacillus simulans. Eight mice were in each group. The mice were fed the MCD diet for 30 days, and every 3 days were administered either PBS (200 μL) or mouse-derived Lactobacillus simulans (200 μL) via gavage. Liver tissue was stained with hematoxylin and eosin (HE) to detect changes in liver fat deposition and stem cell damage. Serum ALT and AST levels, and liver triglyceride (TG) content were also measured. Results are as follows: Figure 1 As shown, compared with the MCD+PBS group, the MCD+mouse combined with Lactobacillus group showed significantly reduced fat deposition and hepatocyte damage in the liver tissue of mice (Figure A, P < 0.05); compared with the MCD+PBS group, the serum ALT and AST levels in mice in the MCD+mouse combined with Lactobacillus group were significantly reduced (Figure B, P < 0.05); compared with the MCD+PBS group, the triglyceride (TG) content in the liver tissue of mice in the MCD+mouse combined with Lactobacillus group was decreased (Figure C, P < 0.05).

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

Claims

1. A strain of *Lactobacillus assemblica* ( Ligilactobacillus murinus It is deposited at the China Center for Type Culture Collection, with accession number CCTCC M 20242901.

2. A medicine for the prevention or treatment of disease, characterized in that, The drug comprises the *Lactobacillus murineis* strain or its culture as described in claim 1, wherein the disease is non-alcoholic fatty liver disease induced by choline methionine deficiency.

3. The drug according to claim 2, characterized in that, The unit dose of the drug contains 1×10 9 Mouse-associated lactobacillus with a CFU or higher.

4. The drug according to claim 2, characterized in that, The drug is a lyophilized powder or tablet.

5. The drug according to claim 2, characterized in that, The drug contains pharmaceutically acceptable excipients.

6. The drug according to claim 5, characterized in that, The excipients include carriers, and / or diluents, and / or excipients.

7. The application of the *Lactobacillus murineis* as described in claim 1, characterized in that, The application is for the preparation of drugs to prevent or treat a disease, namely, non-alcoholic fatty liver disease induced by choline methionine deficiency.

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

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

Images