Application of agonist targeting LXR alpha in liver macrophages

Abstract

The invention discloses application of an agonist targeting LXRalpha in liver macrophages. The relationship between the LXRalpha in the liver macrophages and dyslipidemia, chronic hepatitis B, liver fibrosis and liver tumor is researched. According to the present invention, the HBV infection with the chronic hepatitis B can cause the abnormal lipid metabolism of the intrahepatic macrophages, the enhancement of the LXR alpha expression can inhibit the abnormal lipid metabolism of the intrahepatic macrophages so as to control the malignant progression of the chronic hepatitis B, and the LXR alpha agonist inhibits the formation of liver fibrosis and tumor stem cells by inhibiting the LXR alpha-mediated abnormal lipid metabolism of the intrahepatic macrophages so as to control the chronic hepatitis B malignant progression; a novel possible treatment means is provided for treatment of chronic hepatitis B, hepatic fibrosis and tumors.

Description

Technical Field

[0001] The present invention relates to the field of biological and medical technology, and in particular to the application of an agonist targeting LXRα in liver macrophages in lipid metabolism abnormalities, liver fibrosis, liver tumor stemness, etc. Background Art

[0002] The harm of hepatitis B virus (HBV) infection lies in that its persistent infection can initiate the dynamic progression of liver fibrosis, cirrhosis, and liver cancer. Currently, there are about 350-400 million chronic hepatitis B virus carriers in the world, and 500,000-1 million patients die from chronic hepatitis B-related cirrhosis, hepatocellular carcinoma (HCC), and liver failure each year. 1,2 . At present, there are as many as 93 million patients with chronic hepatitis B infection in my country. It is predicted that by 2050, even under an ideal model, the number of chronic hepatitis B patients in my country will still exceed 27 million. Lamivudine and adefovir, which are currently used for hepatitis B virus, are mainly designed based on nucleoside analogs to inhibit the replication of the viral genome in the body, aiming to reduce the virus content in the blood to below the detection limit. However, this therapy has limited efficacy, large side effects, and cannot completely eliminate covalently closed circular DNA (cccDNA) in the liver. In addition, there is no specific drug for hepatitis B virus. Therefore, in addition to the hepatitis B virus itself as a target, there is an urgent need to find potential host targets, mobilize the body's immune response, and provide new treatment strategies to assist in achieving functional cure of HBV.

[0003] Clinically, more than half of HCCs are caused by chronic HBV infection. Studies have shown that the initiation of liver cancer is mediated by a small group of stem cell-like liver cancer cells. 3 The presence of some tumor cells with the potential for self-renewal, proliferation and multidirectional differentiation is associated with the initiation, metastasis and recurrence of liver cancer. Yamanaka factors are a group of transcription factors that can induce the generation of pluripotent stem cells and also play an important role in promoting the formation of tumor stem cells. 4 The tumor cell sphere assay (tumor sphere size and number) is an important method for in vitro identification of tumor stem cells and is also the gold standard for measuring the stemness of tumor cells. If the stem cell-like cancer cell population can be targeted and eliminated, then this approach is expected to become one of the reliable strategies for the treatment of liver malignancies.

[0004] Studies have shown that HBV regulates host immune escape to prevent viral clearance. HBV inhibits the production of antiviral cytokines in liver macrophages, thereby establishing persistent HBV infection 5In addition, HBV reprogrammed the metabolic level of macrophages, enhanced oxidative phosphorylation, and inhibited the antiviral response of M1-like macrophages. 6 Studies have shown that HBV proteins inhibit the migration of monocytes and the respiratory burst of neutrophils, thereby suppressing the innate immune response 7 . Therefore, disturbances in metabolic and immune homeostasis in the host liver are important manifestations of HBV infection. Lipid metabolism has been assigned a key role in shaping Kupffer cell function 8–11 . It has been reported that scar-related TREM + CD9 + Liver macrophages have a profibrotic effect. HBeAg, as a ligand for Toll-like receptor-2 (TLR2), activates macrophages and exacerbates liver fibrosis by promoting the paracrine effect of macrophages and hepatic stellate cells. 12 In addition, HCV induces abnormal cholesterol metabolism in macrophages, promoting fatty hepatitis and liver fibrosis 13 . The above studies all indicate that liver macrophages in pathological conditions can promote disease progression. Therefore, liver macrophages are likely to have a potential role in the chronicity and malignancy of hepatitis B. However, no studies have reported whether abnormal lipid metabolism in hepatic macrophages promotes the progression of chronic hepatitis B, and chronic hepatitis B therapy based on regulating macrophage lipid metabolism is also in urgent need of research.

[0005] Liver X receptor (LXR) is one of the most important nuclear receptors in the human body. It is divided into two subtypes, LXRα and LXRβ. LXRα is mainly expressed in tissues with active lipid metabolism, such as hepatocytes, macrophages and adipocytes, and is activated by cholesterol and its derivatives as ligands. 14 . After LXRα binds to the ligand and is activated, it binds to the corresponding cis-acting elements of downstream genes (such as Abca1, Abcg1, Apoe, Cyp7a1, etc.), upregulates the transcriptional expression of genes, and plays an important role in cholesterol metabolism and fatty acid synthesis. At present, LXRα agonists are mainly used in the treatment of various clinical glucose and lipid metabolism-related diseases. At present, LXRα agonists have not been used in the treatment of chronic hepatitis B. Whether LXRα is involved in regulating the progression of chronic hepatitis B, as well as its cellular and biological mechanisms, need further study.

[0006] The chronic hepatitis B mouse model constructed by AAV-HBV viral vector can continuously produce HBV viral particles and HBV antigens, induce long-term HBV viremia and antigenemia, and cause liver fibrosis and liver cancer, making it the main model for studying the immune mechanism of chronic hepatitis B and evaluating the efficacy. This study combined single-cell transcriptome research on the intrahepatic microenvironment of HBV-related liver cancer and confirmed that intrahepatic inflammation, especially inflammatory cells, play a key role in promoting disease progression. Therefore, determining key effector cells and inflammatory targets is the basis for the implementation of the "targeted inflammation" strategy in the treatment of chronic hepatitis B.

[0007] In summary, it is very necessary and important to find new effective therapeutic targets for dyslipidemia, chronic hepatitis B, liver fibrosis and liver cancer.

[0008] References

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[0018] 10.Lee,M.-S.,and Bensinger,S.J.(2022).Reprogramming cholesterolmetabolismin macrophages and its role in host defense against cholesterol-dependent cytolysins.Cell.Mol.Immunol.19,327–336.https: / / doi.org / 10.1038 / s41423-021-00827-0.

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[0022] 14. Zelcer, N., and Tontonoz, P. (2006). Liver X receptors as integrators of metabolic and inflammatory signaling. J. Clin. Invest. 116, 607–614. https: / / doi.org / 10.1172 / JCI27883. Summary of the invention

[0023] In view of the defects in the existing treatment technologies for chronic hepatitis B and liver tumors, the present invention studies the relationship between LXRα in liver macrophages and abnormal lipid metabolism, chronic hepatitis B, liver fibrosis and liver tumors, and confirms the possibility that LXRα agonists in liver macrophages can be used as new therapeutic drugs for chronic hepatitis B, liver fibrosis and liver tumors.

[0024] In order to achieve the above object, the present invention adopts the following technical solutions:

[0025] First, we studied the relationship between intrahepatic macrophage metabolism and the progression of chronic hepatitis B

[0026] A mouse chronic hepatitis B model was constructed. The Kupffer cells of mice in the chronic hepatitis B model group contained a large amount of neutral lipids, and the content was significantly higher than that in the control group mice. HBV infection with chronic hepatitis B can cause abnormal lipid metabolism in intrahepatic macrophages.

[0027] Secondly, we studied the relationship between LXRα expression and abnormal lipid metabolism in Kupffer cells.

[0028] Enhancing LXRα expression can inhibit abnormal lipid metabolism in Kupffer cells.

[0029] Third, study the relationship between LXRα and liver fibrosis and liver tumors

[0030] GW3965, T0901317, AZ876, acetylrohanic anhydride and 22(R)-hydroxycholesterol inhibit liver fibrosis and tumor formation by downregulating LXRα-mediated abnormal lipid metabolism in intrahepatic macrophages, thereby controlling the progression of chronic hepatitis B.

[0031] Compared with the prior art, the beneficial effects of this application are:

[0032] 1. It was determined that the effector cells of chronic hepatitis B progression (acute inflammation, chronic inflammation, liver fibrosis and liver cancer) are liver macrophages (Kupffer cells);

[0033] 2. Enhancing LXRα expression can inhibit abnormal lipid metabolism in Kupffer cells;

[0034] 3. It was confirmed that LXRα in liver macrophages is an inflammatory target for controlling the progression of chronic hepatitis B, which laid the foundation for the implementation of the "targeted inflammation" strategy in the treatment of chronic hepatitis B;

[0035] 4. It was determined that stimulating the expression of LXRα in macrophages reversed liver fibrosis and tumor formation by reversing abnormal lipid metabolism in Kupffer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 , the mouse chronic hepatitis B model simulates the natural progression of human chronic hepatitis B cancer transformation, where A is the result diagram of the acute inflammation stage, chronic inflammation stage and liver fibrosis stage,

[0037] B is the result diagram of liver cancer stage;

[0038] Figure 2 , are the observation results of macrophages, among which,

[0039] A is the GO analysis of liver macrophages.

[0040] B is the oil red staining result of macrophages in the liver of model mice.

[0041] C is the oil red staining result of macrophages after co-culture of Hep AD38 and macrophages;

[0042] Figure 3 , is the in vitro detection of LXRα expression in macrophages, among which,

[0043] A shows the expression of LXRα in macrophages in the co-culture system after tetracycline withdrawal. The expression level of LXRα in macrophages co-cultured with HepAD38 cells (HBV-positive) from which tetracycline was withdrawn was significantly reduced.

[0044] B shows that after adding two LXR agonists, GW3965 and T0901317, respectively, into the co-culture system, the protein levels of macrophage LXRα and its downstream molecule ABCA1 increased significantly.

[0045] C shows the changes in lipid droplets in macrophages after adding two LXR agonists, GW3965 and T0901317, respectively, to the co-culture system. After adding LXR agonists, the content of lipid droplets in macrophages was significantly reduced;

[0046] Figure 4 , is the result of detecting the expression level of liver fibrosis markers (TGF-β and MMP-2) in hepatocytes after co-culture with macrophages with normal or abnormal lipid metabolism;

[0047] Figure 5 , The expression level detection results of tumor stemness markers (Yamanaka factors: NANOG, KLF4, OCT4, cMYC, LIN28A) in hepatocytes after co-culture with macrophages with normal or abnormal lipid metabolism;

[0048] Figure 6 , are representative results of stem cell spheres in the hepatocyte sphere formation experiment after co-culture with macrophages with normal or abnormal lipid metabolism;

[0049] Figure 7 , is the result of the decrease in the expression level of fibrosis markers (TGF-β and MMP-2) in hepatocytes after adding LXRα agonist GW3965 into the co-culture system;

[0050] Figure 8 , is the result of the decrease in the expression level of tumor stemness markers (Yamanaka factors: NANOG, KLF4, OCT4, cMYC, LIN28A) in hepatocytes after the addition of LXRα agonist GW3965 to the co-culture system;

[0051] Fig. 9 , is the number of microspheres of stem cell spheres in the hepatocyte sphere formation experiment after co-culture with macrophages with normal or abnormal lipid metabolism. The number and diameter of hepatocyte spheres induced by macrophages with abnormal lipid metabolism increased;

[0052] Fig.10 , AZ876, acetylrohanic anhydride and 22(R)-hydroxycholesterol and the expression level of LXRα in macrophages;

[0053] Fig.11 , AZ876, acetylrohanic anhydride and 22(R)-hydroxycholesterol and the expression levels of liver fibrosis markers and cancer stem cell markers;

[0054] Fig.12 , the number of LXRα-positive Kupffer cells in normal tissues, paracancerous tissues, and cancer tissues.

[0055] in Figure 1-Figure 12 middle,

[0056] *, **, *** and **** represent P<0.05, P<0.01, P<0.001 and P<0.0001, respectively, indicating that the difference between the two groups was statistically significant. DETAILED DESCRIPTION

[0057] The technical scheme of the present invention is specifically and detailedly described below in conjunction with the embodiments of the present invention and the accompanying drawings. It should be noted that the following embodiments are only used to illustrate the present invention and are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0058] Experimental Materials

[0059]

[0060]

[0061] 2. Instruments

[0062] Instrument Name model Manufacturer Mouse Holder / Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. Infinite 200 Microplate Reader Infinite 200 TECAN Switzerland EVOS M7000 Imaging System EVOS M7000 Thermo Fisher Scientific Electrophoresis tank 1658001 Bio-Rad Corporation Transfer membrane 1703935 Bio-Rad Corporation Luminescence imaging system VILBER FUSION FX7 Vilber, France 7500 Real-time Fluorescence Quantitative PCR System ABI 7500 Thermo Fisher Scientific

[0063] 3. Preparation of serum-free stem cell culture medium for stem cell sphere formation experiment

[0064] Take the preparation of 100mL culture medium as an example:

[0065] Reagents volume DMEM / F12 97mL B27 additive (50×) 2mL N2 additive (100×) 1mL EGF (200 μg / mL) 10μL FGFb (200 μg / mL) 10μL Double antibody (100×) 100μL Total 100mL

[0066] Example 1: Construction of a mouse chronic hepatitis B model

[0067] 1. Methods

[0068] 1. Inject 1×10 11 The AAV-HBV D type vector and control vector packaged by GC lentivirus were used to construct a mouse chronic hepatitis B model and a control mouse model to simulate the natural progression of human chronic hepatitis B cancer transformation and compared with control mice.

[0069] 2. Results

[0070] The mice underwent acute inflammation, chronic inflammation, and liver fibrosis ( Figure 1 A) Liver cancer stage ( Figure 1B). This indicates that the AAV8-HBV virus in this study can successfully enter and infect mouse liver tissue through tail vein injection, leading to viremia and liver inflammation, providing a good animal model for the subsequent establishment of single-cell transcriptome maps and mechanism exploration.

[0071] Example 2: Relationship between Kupffer cell lipid metabolism function and progression of chronic hepatitis B

[0072] 1. Methods

[0073] The differentially expressed genes and functional enrichment (GO analysis) of intrahepatic macrophages (Kupffer cells) in the single-cell data of intrahepatic immune cells of mice at different stages after chronic hepatitis B infection were searched, and the functional differences between Kupffer cells in the hepatitis B infection group and the control group were analyzed.

[0074] 2. Results

[0075] It was found that with the progression of chronic hepatitis B, the lipid metabolism function of Kupffer cells was significantly abnormal ( Figure 2 A). This study further isolated Kupffer cells from the livers of AAV-HBV model mice and control mice, and used Oil Red O staining to observe neutral lipids in the cells. The results are shown in Figure 2. Figure 2 As shown in B, the Kupffer cells of mice in the chronic hepatitis B model group contained a large amount of neutral lipids, and the content was significantly higher than that in the control group mice.

[0076] 3. Analysis and conclusion

[0077] Abnormal lipid metabolism in Kupffer cells is one of the important signs of chronic hepatitis B, and it also suggests that abnormal lipid metabolism function mediated by Kupffer cells is likely to be involved in the progression of chronic hepatitis B.

[0078] Example 3: Relationship between HBV infection, chronic hepatitis B and abnormal lipid metabolism in hepatic macrophages

[0079] 1. Methods

[0080] In vitro, THP-1 cells were first induced by phorbol ester (PMA) for 24 hours to obtain macrophages (M0). The Hep AD38 cell line is a human cell line that can inducibly express HBV and express HBV when tetracycline (Tet-) is withdrawn. M0 macrophages were co-cultured with HepAD38 cells expressing or not expressing HBV in a Transwell chamber for 48 hours, and then the morphological changes of macrophages were observed. The macrophages were stained with an Oil Red O staining kit to observe the intracellular lipid droplet content.

[0081] 2. Results

[0082] After co-culturing M0 with Hep AD38 (Tet-), lipid droplet aggregation was found in macrophages. In vitro co-culture experiments were performed using THP-1-derived macrophages and HepAD38 cell lines (Figure), and it was found that a large amount of neutral lipids ( Figure 2 C).

[0083] 3. Analysis and conclusion

[0084] The above results confirm from the perspective of in vitro experiments that HBV + Hepatocytes do cause abnormal lipid metabolism in hepatic macrophages, and HBV + The cross-talk between hepatocytes and Kupffer cells is crucial.

[0085] Example 4: Relationship between LXR α expression and abnormal lipid metabolism in Kupffer cells

[0086] Further detection of LXRα expression revealed that LXRα, which is closely related to lipid metabolism, was significantly decreased in the macrophages of the co-culture system ( Figure 3 A); After adding two LXR agonists, GW3965 and T0901317, the expression of LXRα and its downstream molecule ABCA1 in macrophages was significantly increased ( Figure 3 B), lipid droplets were significantly reduced ( Figure 3 C). This indicates that abnormal lipid metabolism of Kupffer cells is mediated by LXRα, and LXRα agonists can alleviate abnormal lipid metabolism of Kupffer cells. Example 5 illustrates in detail that abnormal lipid metabolism of Kupffer cells affects the chronicity and malignancy of chronic hepatitis B (CHB).

[0087] Example 5: Relationship between LXRα and liver fibrosis and liver tumors

[0088] Co-culture of macrophages with abnormal lipid metabolism with Hep AD38 cells can cause a significant increase in liver fibrosis markers (TGF-β, MMP-2) in hepatocytes ( Figure 4 ). The tumor cell sphere formation experiment (tumor sphere size and number) is an important method for in vitro identification of tumor stem cells and is also the gold standard for measuring the stemness of tumor cells. The study found that co-culture of macrophages with abnormal lipid metabolism and Hep AD38 cells can increase the tumor stem cell formation ability of Hep AD38 cells ( Figure 5 ), which is manifested by an increase in the number and diameter of microspheres ( Figure 6 ); the addition of LXRα agonist (GW3965) can significantly reverse the expression of fibrosis-related marker molecules ( Figure 7 ) and liver cancer stem cell marker( Figure 8 ) level, reducing the number and diameter of tumor stem cell spheres ( Fig. 9 ). It was confirmed that stimulating the expression of LXRα in macrophages can reverse liver fibrosis and liver tumor formation. It shows that liver fibrosis and tumor formation can be reversed by reversing the abnormal lipid metabolism of Kupffer cells. In short, the application of LXRα agonists to macrophages can target the reduction of the expression of liver fibrosis and tumor stem cell markers, and eliminate the stem cell-like liver cancer cell population. This approach is expected to become one of the strategies for treating CHB-related liver malignancies.

[0089] Example 6: Relationship between LXRα and liver fibrosis and liver tumors

[0090] 1. Methods

[0091] After macrophages were obtained from THP-1 cells induced by PMA in vitro, 9 agonists were added, namely AZ876 (2-tert-butyl-5-phenyl-4-[[4-(1-piperidinyl)phenyl]amino]-3(2H)-isothiazolinone 1,1-dioxide), Iristectorigenin B, Yamogenin, Saikosaponin A, Acetylrohanic Anhydride (APD), BI456906 (Survodutide), Semaglutide, Nagilactone B, and 22(R)-hydroxycholesterol (22(R)-Hch), and the culture was continued for 48 hours.

[0092] 2. Results

[0093] It was found that AZ876, acetylrohanic anhydride and 22(R)-hydroxycholesterol could significantly increase the expression level of LXRα in macrophages ( Fig.10 In the co-culture system, the three agonists were used to treat macrophages with abnormal lipid metabolism, and then the macrophages were co-cultured with HepAD38 cells, which significantly inhibited the expression levels of liver fibrosis markers and tumor stem cell markers ( Fig.11 ).

[0094] 3. Analysis and conclusion

[0095] AZ876, acetylrohanic anhydride and 22(R)-hydroxycholesterol inhibit liver fibrosis and liver tumor formation by regulating LXRα downregulation-mediated abnormal lipid metabolism in Kupffer cells.

[0096] Example 7: Relationship between LXRα expression level in Kupffer cells and HBV-related liver cancer

[0097] 1. Methods

[0098] The liver tissue microarray of HBV-related liver cancer patients was analyzed using multicolor immunohistochemistry, and the number of LXRα-positive Kupffer cells in different regions was counted.

[0099] 2. Results

[0100] The results are as follows Fig.12 As shown, the number of LXRα-positive Kupffer cells was highest in normal tissues, followed by paracancerous tissues, and lowest in cancer tissues.

[0101] 3. Analysis and conclusion

[0102] These results indicate that the expression level of LXRα in Kupffer cells is closely related to the occurrence and development of HBV-related liver cancer.

[0103] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

[0104] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. Use of 3-[3-[[2-chloro-3-(trifluoromethyl)phenyl]methylamino]propoxy]phenylacetic acid hydrochloride, i.e. GW3965, an agonist of LXRα in liver macrophages, in the preparation of a drug for treating abnormal lipid metabolism in liver macrophages.

2. Use of N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]benzenesulfonamide, i.e. T0901317, an agonist of LXRα in liver macrophages, in the preparation of a drug for treating abnormal lipid metabolism in liver macrophages.

3. Use of 3-[3-[[2-chloro-3-(trifluoromethyl)phenyl]methylamino]propoxy]phenylacetic acid hydrochloride, GW3965, an agonist of LXRα in liver macrophages, in the preparation of a drug for controlling the progression of chronic hepatitis B, characterized in that: The drug is achieved by inhibiting abnormal lipid metabolism of liver macrophages, thereby reducing liver fibrosis and inhibiting the formation of liver tumors.

4. The use of N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]benzenesulfonamide, i.e. T0901317, an agonist of LXRα in liver macrophages, in the preparation of a drug for controlling the progression of chronic hepatitis B, characterized in that: The drug is achieved by inhibiting abnormal lipid metabolism of liver macrophages, thereby reducing liver fibrosis and inhibiting the formation of liver tumors.

5. Use of 3-[3-[[2-chloro-3-(trifluoromethyl)phenyl]methylamino]propoxy]phenylacetic acid hydrochloride, GW3965, an agonist of LXRα in liver macrophages, in the preparation of a drug for reducing liver fibrosis, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

6. Use of N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]benzenesulfonamide, T0901317, an agonist of liver macrophage LXRα, in the preparation of a drug for reducing liver fibrosis, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

7. Use of 3-[3-[[2-chloro-3-(trifluoromethyl)phenyl]methylamino]propoxy]phenylacetic acid hydrochloride, GW3965, an agonist of LXRα in liver macrophages, in the preparation of a drug for inhibiting liver tumor formation, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

8. Use of N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]benzenesulfonamide, T0901317, an agonist of LXRα in liver macrophages, in the preparation of a drug for inhibiting liver tumor formation, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

9. Use of 2-tert-butyl-5-phenyl-4-[[4-(1-piperidinyl)phenyl]amino]-3(2H)-isothiazolinone 1,1-dioxide, AZ876, an agonist of LXRα in liver macrophages, in the preparation of a drug for reducing liver fibrosis, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

10. Use of acetylrohanic anhydride, an agonist of LXRα in liver macrophages, in the preparation of a drug for reducing liver fibrosis, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

11. Use of 22(R)-hydroxycholesterol, an agonist of LXRα in liver macrophages, in the preparation of a drug for reducing liver fibrosis, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

12. Use of 2-tert-butyl-5-phenyl-4-[[4-(1-piperidinyl)phenyl]amino]-3(2H)-isothiazolinone 1,1-dioxide, AZ876, an agonist of LXRα in liver macrophages, in the preparation of a drug for inhibiting liver tumor formation, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

13. Use of acetylrohanic anhydride, an agonist of LXRα in liver macrophages, in the preparation of a drug for inhibiting liver tumor formation, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

14. Use of 22(R)-hydroxycholesterol, an agonist of LXRα in liver macrophages, in the preparation of a drug for inhibiting liver tumor formation, characterized in that: The drug works by treating abnormal lipid metabolism in liver macrophages.

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