Application of AIM2 inhibitor in preparation of medicine for treating hepatic fibrosis

Abstract

The invention provides application of an AIM2 inhibitor in preparation of a medicine for treating hepatic fibrosis. And the AIM2 inhibitor is ODN A151. Researches show that ODN A151 as an effective component of the medicine for treating hepatic fibrosis can relieve the degree of structural disorder and inflammatory cell infiltration of hepatic fibrosis mouse liver tissues, reduce expression of pyroptosis related molecules GSDMD and proinflammatory cytokines IL-1beta and IL-18, and reduce expression of mRNA of hepatic fibrosis marker molecules COL1A1 and alpha-SMA. The application proves that the ODN A151 can be used for treating the hepatic fibrosis, provides a new theoretical basis and experimental basis for the development of new medicines for clinically preventing or reversing the progress of the hepatic fibrosis, and has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of an AIM2 inhibitor in the preparation of a drug for treating liver fibrosis. Background Art

[0002] Hepatic fibrosis (HF) refers to the excessive deposition of diffuse extracellular matrix (especially collagen) in the liver. Hepatic stellate cells are the main source of diffuse extracellular matrix in the liver, and the activation or transdifferentiation of hepatic stellate cells into myofibroblasts is the main driving factor of liver fibrosis. Hepatic fibrosis is an abnormal repair response to chronic liver damage caused by various factors, such as viral hepatitis, alcoholic liver disease, alcoholic fatty liver disease or autoimmune liver disease, which can lead to chronic liver injury inflammation and fibrosis. Without effective intervention, advanced liver fibrosis will further develop into irreversible cirrhosis and even liver cancer, which is the main cause of liver-related death worldwide. It is estimated that there are about 70 million patients with liver fibrosis in my country. Liver fibrosis has seriously endangered people's health. The treatment of liver fibrosis aims to prevent or reverse the progression of liver fibrosis, improve the liver function and structure of patients, delay the occurrence of cirrhosis and its decompensation stage, improve the quality of life of patients and prolong their survival. However, there is currently no nationally approved drug that can effectively treat and improve liver fibrosis. Therefore, it is particularly urgent to find drugs to treat liver fibrosis. Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides the use of an AIM2 inhibitor in the preparation of a drug for treating liver fibrosis, wherein the AIM2 inhibitor is ODN A151. The present invention proves that ODN A151 can inhibit liver fibrosis, so using ODN A151 in the preparation of a drug for treating liver fibrosis provides a new theoretical basis and experimental basis for the clinical development of new drugs to prevent or reverse the progression of liver fibrosis, and has good application prospects.

[0004] The first aspect of the present invention provides use of an AIM2 inhibitor in the preparation of a drug for treating liver fibrosis, wherein the AIM2 inhibitor is ODN A151.

[0005] Melanoma deficiency factor 2 (AIM2) is an intracellular DNA sensor and one of the important pattern recognition receptors that make up the inflammasome. It plays an important role in the innate immune response. AIM2 consists of an N-terminal pyrin domain (PYD) and a C-terminal hematopoietic interferon-induced nuclear protein (HIN200) domain with a 200 amino acid repeat sequence. It is mainly located in the cytoplasm and belongs to the PYHIN protein family. Among them, HIN-200 directly binds to double-stranded DNA (dsDNA), and PYD recruits the protein apoptosis-associated speck-like protein (ASC) and caspase-1 precursor to form the AIM2 inflammasome. Inflammasome-dependent cell pyroptosis is a new type of inflammatory cell death that is involved in the progression of many diseases such as infection, metabolic disorders and cancer. The process of AIM2 inflammasome-dependent cell pyroptosis is as follows: AIM2 is a key sensor in the AIM2 inflammasome, which can recognize double-stranded DNA (dsDNA) and trigger the cascade reaction of the AIM2 inflammasome, thereby inducing cell pyroptosis. After cell pyroptosis, a large amount of proinflammatory cytokines (interleukin 1β (IL-1β) and interleukin 18 (IL-18)) can be released. The massive release of proinflammatory cytokines will further promote the development of inflammation.

[0006] Through a large number of studies, the inventors found that the increased expression of proinflammatory cytokines (interleukin 1β (IL-1β) and interleukin 18 (IL-18)) of AIM2 inflammasome-dependent cell pyroptosis and cell pyroptosis-related molecules (such as pyrocontin D (GSDMD)) would aggravate the development of liver fibrosis.

[0007] Based on a lot of previous exploration, the inventors found that ODN A151 can be used as a single drug, and has a very obvious therapeutic effect on the treatment of liver fibrosis, with significant effects. The inventors' research shows that ODN A151 can reduce the expression of GSDMD, IL-1β, IL-18, and reduce the expression of liver fibrosis marker molecules COL1A1 and α-SMA. Therefore, ODN A151 is used to prepare drugs for the treatment of liver fibrosis.

[0008] It is understood that the actual dosage of ODN A151 administered in the present invention may vary greatly depending on a variety of factors, such as the target cell, type of organism or tissue thereof, the general condition of the subject to be treated, the route of administration, the mode of administration, and the like.

[0009] Furthermore, the drug has ODN A151 as the only active ingredient.

[0010] The drug for treating liver fibrosis of the present invention uses ODN A151 as an active ingredient and can inhibit liver fibrosis.

[0011] Furthermore, the medicine also includes excipients acceptable in pharmacy, food science or health care products science.

[0012] The present invention does not specifically limit the above-mentioned adjuvants, and suitable pharmaceutical preparations can be selected according to the selection of specific dosage forms. Examples of available adjuvants include excipients (e.g., sugar derivatives such as sucrose, lactose, honey, glucose, mannitol and sorbitol; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, etc.), binders (e.g., gelatin, polyvinyl pyrrolidone and polyethylene glycol), lubricants (e.g., talc, calcium stearate, magnesium stearate, spermaceti, boric acid, etc.), stabilizers (methyl parahydroxybenzoate, propyl parahydroxybenzoate, etc.), etc.

[0013] Furthermore, the ODN A151 is used to prepare a drug for reducing the expression of liver fibrosis marker molecules COL1A1 and α-SMA to inhibit liver fibrosis.

[0014] Furthermore, the ODN A151 is used to prepare a drug for reducing the expression of the cell pyroptosis-related molecule GSDMD and inhibiting liver fibrosis.

[0015] Furthermore, the ODN A151 is used to prepare a drug for reducing the expression of pro-inflammatory cytokines IL-1β and IL-18 and improving liver fibrosis inflammation.

[0016] Furthermore, the ODN A151 is used to prepare a drug for reducing the degree of liver tissue structural disorder and inflammatory cell infiltration.

[0017] Furthermore, the ODN A151 is used to prepare a lead drug for inhibiting carbon tetrachloride-induced liver fibrosis mouse model.

[0018] The present invention does not limit the specific preparation form of the drug, and the specific preparation form can be selected according to needs. Further, the dosage form of the drug can be any one of injection, capsule, tablet, microcapsule preparation, suppository, solution, powder or spray.

[0019] The second aspect of the present invention provides a preparation of a drug for treating liver fibrosis, wherein the therapeutically active ingredient includes the ODN A151.

[0020] In summary, the solution of the present invention has at least the following effects:

[0021] The present invention develops a new use of the AIM2 inhibitor ODN A151, and uses ODN A151 in the preparation of drugs for treating liver fibrosis. Studies have shown that ODN A151, as an effective ingredient of drugs for treating liver fibrosis, can reduce the structural disorder and inflammatory cell infiltration of liver tissue in mice with liver fibrosis, reduce the expression of cell pyroptosis-related molecules GSDMD, proinflammatory cytokines IL-1β and IL-18, reduce the expression of liver fibrosis marker molecules COL1A1 and α-SMA, inhibit hepatic stellate cell activation, and reduce collagen deposition in liver tissue. The present invention confirms that ODN A151 can be used to treat liver fibrosis, provides a new theoretical basis and experimental basis for the clinical development of new drugs to prevent or reverse the progression of liver fibrosis, and has good application prospects.

[0022] The present invention uses the expression of pro-inflammatory cytokines IL-1β and IL-18 of AIM2 inflammasome-dependent cell pyroptosis and cell pyroptosis-related molecule GSDMD in liver tissues of patients with liver fibrosis and liver tissues of normal healthy people. The results show that the expression of GSDMD, IL-1β and IL-18 in liver tissues of patients with liver fibrosis is significantly higher than that in liver tissues of normal healthy people. It shows that AIM2 inflammasome-dependent cell pyroptosis occurs in patients with liver fibrosis, and the occurrence of AIM2 inflammasome-dependent cell pyroptosis will aggravate the development of liver fibrosis, and it is verified by constructing a CCl4-induced liver fibrosis mouse model. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] 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, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 Immunofluorescence (IF) detection was performed on liver tissue samples of patients with liver fibrosis and normal healthy subjects (healthy controls).

[0025] Figure 2 Immunohistochemistry (IHC) was performed on liver tissue samples from patients with liver fibrosis and normal healthy subjects (healthy controls).

[0026] Figure 3 The liver tissue sections obtained from the blank control group and the model group were stained with hematoxylin-eosin (HE).

[0027] Figure 4The liver tissue sections obtained from the blank control group and the model group were stained with α-smooth muscle actin (α-SMA).

[0028] Figure 5 The liver tissue sections obtained from the blank control group and the model group were stained with Sirius Red.

[0029] Figure 6 Immunofluorescence (IF) detection was performed on the liver tissue sections obtained from the blank control group and model group.

[0030] Figure 7 Immunohistochemistry (IHC) detection was performed on the liver tissue sections obtained from the blank control group and model group.

[0031] Figure 8 The liver tissue sections obtained from the blank control group, model group, and ODN A151 treatment group were stained with hematoxylin-eosin (HE).

[0032] Fig. 9 Immunohistochemistry (IHC) detection was performed on the liver tissue sections obtained from the blank control group, model group, and ODN A151 treatment group.

[0033] Fig.10 Real-time fluorescence quantitative PCR (qRT-PCR) detection was performed on the liver tissues of the blank control group, model group and ODN A151 treatment group.

[0034] Fig.11 The liver tissue sections obtained from the blank control group, model group, and ODN A151 treatment group were stained for α-smooth muscle actin (α-SMA).

[0035] Fig.12 The liver tissue sections obtained from the blank control group, model group, and ODN A151 treatment group were stained with Sirius Red. DETAILED DESCRIPTION

[0036] To make the purpose, technical scheme and advantages of the present invention clearer, the technical scheme in the embodiment of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the art without making creative work are within the scope of protection of the present invention. If the specific technology or conditions are not indicated in the embodiments, the technology or conditions described in the literature in this area or the product instructions are carried out. The reagents or instruments used that do not indicate the manufacturer are all conventional products that can be obtained commercially.

[0037] As used herein, the term "treat" refers to ameliorating a disease or condition (i.e., slowing down or preventing or alleviating the development of a disease or at least one of its clinical symptoms) in some embodiments. In other embodiments, "treat" refers to alleviating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treat" refers to regulating a disease or condition physically (e.g., stabilizing a perceptible symptom) or physiologically (e.g., stabilizing a physical parameter), or both. In other embodiments, "treat" refers to preventing or delaying the onset, occurrence, or worsening of a disease or condition.

[0038] The application of the AIM2 inhibitor provided by the present invention in the preparation of a drug for treating liver fibrosis is described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0039] Sources of Liver Tissue:

[0040] Liver tissue samples were collected from 15 patients with clinically diagnosed liver fibrosis and 9 normal healthy subjects. Written informed consent was obtained from all participants, and the protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Chongqing Medical University (No. 2023-019).

[0041] Example 1 Effect of AIM2 inflammasome-dependent pyroptosis on the progression of liver fibrosis

[0042] 1. Analysis of AIM2 expression in liver tissues of patients with liver fibrosis

[0043] In order to explore the relationship between AIM2 and liver fibrosis, the liver tissue samples of patients with liver fibrosis and healthy subjects (healthy controls) were subjected to immunofluorescence (IF) detection. Figure 1 shown.

[0044] Depend on Figure 1It can be seen that compared with normal healthy people, the expression of AIM2 in the liver tissue of patients with liver fibrosis is increased, and AIM2 is mainly located in liver macrophages, indicating that AIM2 inflammasome-dependent cell pyroptosis mainly occurs in liver macrophages in the process of liver fibrosis.

[0045] 2. Expression analysis of factors related to AIM2 inflammasome-dependent pyroptosis in liver tissues of patients with liver fibrosis

[0046] Immunohistochemistry (IHC) was performed on liver tissue samples from patients with liver fibrosis and healthy controls. Figure 2 shown.

[0047] Depend on Figure 2 It can be seen that compared with normal healthy people, the expression of proinflammatory cytokines (interleukin 1β (IL-1β) and interleukin 18 (IL-18)) of AIM2 inflammasome-dependent cell pyroptosis and cell pyroptosis-related molecules (pyropoietin D (GSDMD)) in the liver tissue of patients with liver fibrosis was increased, indicating that AIM2 inflammasome-dependent cell pyroptosis occurred in patients with liver fibrosis, and the occurrence of AIM2 inflammasome-dependent cell pyroptosis would aggravate the development of liver fibrosis.

[0048] The above results indicate that AIM2 inflammasome-dependent cell pyroptosis occurs in the liver tissue of patients with liver fibrosis, and mainly occurs in liver macrophages.

[0049] Example 2 Construction of CCl4-induced liver fibrosis mouse model

[0050] 1. Experimental Materials

[0051] Male C57BL / 6 mice were purchased from the Experimental Animal Research Center of Chongqing Medical University.

[0052] 2. Construction of CCl4-induced liver fibrosis mouse model

[0053] Ten male C57BL / 6J mice, 6 weeks old, were randomly divided into two groups, namely blank control group (n=5) and model group (carbon tetrachloride (CCl4) injected alone, n=5). The model group was induced to develop liver fibrosis by intraperitoneal injection of CCl4 for 6 weeks. CCl4 was diluted with olive oil (product number: O108686, Aladdin) at a ratio of 1:5, and 1 μl / g was injected each time, twice a week. The blank control group was not treated.

[0054] All animal experimental procedures were approved by the Animal Ethics Committee of Chongqing Medical University (No. 2023-019).

[0055] 3. Obtaining Mouse Liver Tissue and Preparing Liver Tissue Sections

[0056] 72 hours after the last injection of carbon tetrachloride (CCl4), the mice were killed by cervical dislocation, and the liver tissues were removed and prepared into liver tissue slices. The specific steps are as follows:

[0057] 1. Fixation: Fix liver tissue with 4% paraformaldehyde fixative for 24 hours to obtain fixed tissue;

[0058] 2. Dehydration: Dehydrate the fixed tissue using gradient concentrations of alcohol, and soak it in 70%, 80%, 90%, 95% ethanol, anhydrous ethanol I and anhydrous ethanol II in sequence, soaking it in 70% ethanol for 30 minutes, and soaking it in anhydrous ethanol I and anhydrous ethanol II for 1 hour;

[0059] 3. Transparency: Soak the dehydrated tissue in xylene I for 1 hour and xylene II for 1 hour;

[0060] 4. Wax immersion: After the tissue has become transparent, place it in low-melting-point paraffin for 1 hour and high-melting-point paraffin for 1 hour;

[0061] 5. Embedding: Melt the high melting point wax in the automatic embedding machine in advance and keep it in a molten state. Adjust the wax-impregnated tissue to the state required for slicing and put it into the embedding machine. Pour the high melting point wax and place it on ice to wait for it to cool and solidify.

[0062] 6. Slicing: Set the water temperature of the slide spreading pool to 42°C, the temperature of the slide drying area to 65°C, and pre-cool the wax block in the refrigerator. After trimming the embedded tissue to the required cross-section, adjust the thickness to 3 μm and start slicing to obtain liver tissue slices; put the obtained liver tissue slices into the slide spreading pool to remove wrinkles. After the spreading is completed, use a slide pre-coated with APES to pick up the liver tissue slices, place the liver tissue slices in the slide drying area for 2 hours, and then store them at 4°C.

[0063] The above method was used to obtain liver tissue and liver tissue sections of mice in the blank control group and model group, respectively.

[0064] 4. Hematoxylin-eosin (HE) staining

[0065] The liver tissue sections of the blank control group and the model group obtained above were stained with hematoxylin-eosin (HE), and the results were as follows: Figure 3 shown.

[0066] Depend on Figure 3 It can be seen that compared with the blank control group, the liver tissue sections of the model group had disordered structure, hepatocyte edema and degeneration, and the degree of inflammatory cell infiltration was significantly increased.

[0067] 5. α-Smooth Muscle Actin (α-SMA) Staining

[0068] The liver tissue sections of the blank control group and the model group obtained above were stained with α-SMA to detect the activation of hepatic stellate cells in the liver tissue of mice. The results are as follows: Figure 4 shown.

[0069] Depend on Figure 4 It can be seen that compared with the blank control group, the expression of α-SMA, a marker molecule of hepatic stellate cell activation, in the model group was significantly increased.

[0070] 6. Sirius Red staining

[0071] Sirius Red staining was performed on the liver tissue sections of the blank control group and the model group to detect the collagen deposition in the liver tissue of mice. The results are as follows: Figure 5 shown.

[0072] Depend on Figure 5 It can be seen that compared with the blank control group, the collagen deposition in the liver tissue of mice in the model group was significantly increased.

[0073] VII. Analysis of AIM2 expression in mouse liver tissue

[0074] Immunofluorescence (IF) detection was performed on the liver tissue sections of the blank control group and the model group. The results are as follows: Figure 6 shown.

[0075] Depend on Figure 6 It can be seen that compared with the blank control group, the expression of AIM2 in the liver tissue of mice in the model group increased, and AIM2 was mainly localized in liver macrophages, which further illustrates that AIM2 inflammasome-dependent cell pyroptosis mainly occurs in liver macrophages in the process of liver fibrosis.

[0076] 8. Expression analysis of factors related to AIM2 inflammasome-dependent pyroptosis in mouse liver tissue

[0077] Immunohistochemistry (IHC) was performed on the liver tissue sections of the blank control group and the model group. The results are as follows: Figure 7 shown.

[0078] Depend on Figure 7 It can be seen that compared with the blank control group, the expression of proinflammatory cytokines IL-1β and IL-18 of AIM2 inflammasome-dependent cell pyroptosis and cell pyroptosis-related molecule GSDMD in the liver tissue of mice in the model group were increased, indicating that AIM2 inflammasome-dependent cell pyroptosis occurred in the liver tissue of mice in the model group.

[0079] The above results indicate that a CCl4-induced liver fibrosis mouse model was successfully constructed, and AIM2 inflammasome-dependent cell pyroptosis occurred in the liver tissue of liver fibrosis mice, and mainly occurred in liver macrophages.

[0080] Example 3 Effects of AIM2 inhibitor on mice with liver fibrosis

[0081] 1. Experimental Materials

[0082] AIM2 inhibitor: ODN A151 (CAS No.: 1801724-76-4, purchased from MedChemExpress).

[0083] Male C57BL / 6 mice were purchased from the Experimental Animal Research Center of Chongqing Medical University.

[0084] 2. Construction of mouse model of liver fibrosis

[0085] Male C57BL / 6J mice, 15, 6 weeks old, were randomly divided into 3 groups, namely blank control group (n=10), model group (injected with carbon tetrachloride (CCl4) alone, n=10), and ODN A151 treatment group (n=10). The model group and ODN A151 treatment group were induced to develop liver fibrosis by intraperitoneal injection of CCl4 for 6 weeks. CCl4 was diluted with olive oil (product number: O108686, Aladdin) at a ratio of 1:5, and 1 μl / g was injected each time, twice a week. The blank control group was not treated. At the same time, in the ODN A151 treatment group, mice were treated with ODN A151 (100 μg / mouse) by intraperitoneal injection after each injection of CCl4.

[0086] All animal experimental procedures were approved by the Animal Ethics Committee of Chongqing Medical University (No. 2023-019).

[0087] 3. Obtaining Mouse Liver Tissue and Preparing Liver Tissue Sections

[0088] 72 hours after the last injection of carbon tetrachloride (CCl4), the mice were killed by cervical dislocation, and the liver tissues were removed and prepared into liver tissue slices. The specific steps are as follows:

[0089] 1. Fixation: Fix liver tissue with 4% paraformaldehyde fixative for 24 hours to obtain fixed tissue;

[0090] 2. Dehydration: Dehydrate the fixed tissue using gradient concentrations of alcohol, and soak it in 70%, 80%, 90%, 95% ethanol, anhydrous ethanol I and anhydrous ethanol II in sequence, soaking it in 70% ethanol for 30 minutes, and soaking it in anhydrous ethanol I and anhydrous ethanol II for 1 hour;

[0091] 3. Transparency: Soak the dehydrated tissue in xylene I for 1 hour and xylene II for 1 hour;

[0092] 4. Wax immersion: After the tissue has become transparent, place it in low-melting-point paraffin for 1 hour and high-melting-point paraffin for 1 hour;

[0093] 5. Embedding: Melt the high melting point wax in the automatic embedding machine in advance and keep it in a molten state. Adjust the wax-impregnated tissue to the state required for slicing and put it into the embedding machine. Pour the high melting point wax and place it on ice to wait for it to cool and solidify.

[0094] 6. Slicing: Set the water temperature of the slide spreading pool to 42°C, the temperature of the slide drying area to 65°C, and pre-cool the wax block in the refrigerator. After trimming the embedded tissue to the required cross-section, adjust the thickness to 3 μm and start slicing to obtain liver tissue slices; put the obtained liver tissue slices into the slide spreading pool to remove wrinkles. After the spreading is completed, use a slide pre-coated with APES to pick up the liver tissue slices, place the liver tissue slices in the slide drying area for 2 hours, and then store them at 4°C.

[0095] The above method was used to obtain mouse liver tissues and liver tissue sections from the blank control group, model group, and ODN A151 treatment group.

[0096] 4. Hematoxylin-eosin (HE) staining

[0097] The liver tissue sections of the blank control group, model group, and ODN A151 treatment group obtained above were stained with hematoxylin-eosin (HE), and the results were as follows: Figure 8 shown.

[0098] Depend on Figure 8 It can be seen that compared with the blank control group, hepatocyte degeneration was observed in the model group and the ODN A151 treatment group. The liver tissue structure disorder and inflammatory cell infiltration in the model group were significantly reduced in the ODN A151 treatment group compared with the model group, indicating that ODN A151 can improve liver fibrosis inflammation.

[0099] 5. Immunohistochemistry (IHC) detection

[0100] Immunohistochemistry (IHC) was performed on the liver tissue sections of the blank control group, model group, and ODN A151 treatment group. The results are as follows: Fig. 9 shown.

[0101] Depend on Fig. 9It can be seen that compared with the blank control group, the expressions of proinflammatory cytokines IL-1β and IL-18 and pyroptosis-related molecule GSDMD of AIM2 inflammasome-dependent cell pyroptosis in the liver tissue of mice in the model group were increased, while the expressions of proinflammatory cytokines IL-1β and IL-18 and pyroptosis-related molecule GSDMD of AIM2 inflammasome-dependent cell pyroptosis in the ODN A151 treatment group were significantly decreased compared with the model group, indicating that ODN A151 reduced the occurrence of AIM2 inflammasome-dependent cell pyroptosis and improved liver fibrosis inflammation.

[0102] 5. Real-time fluorescence quantitative PCR (qRT-PCR) detection

[0103] The liver tissues of the blank control group, model group, and ODN A151 treatment group obtained above were subjected to qRT-PCR to detect the expression of liver fibrosis marker molecules COL1A1 and α-SMA in the liver tissues of each group of mice. The specific steps were to extract RNA from the mouse liver tissue according to the instructions of the qRT-PCR kit (9108, Takara, Japan) provided by the manufacturer, and the qRT-PCR results were analyzed. The results are shown in Fig.10 shown.

[0104] Depend on Fig.10 It can be seen that compared with the model group, the mRNA expression of liver fibrosis markers COL1A1 and α-SMA in the liver tissue of mice in the ODN A151 treatment group was significantly reduced, indicating that ODN A151 significantly inhibited liver fibrosis.

[0105] 6. α-Smooth Muscle Actin (α-SMA) Staining

[0106] The liver tissue sections of the blank control group, model group, and ODN A151 treatment group obtained above were stained with α-SMA to detect the activation of hepatic stellate cells in the liver tissue of mice. The results are as follows: Fig.11 shown.

[0107] Depend on Fig.11 It can be seen that compared with the blank control group, the expression of α-SMA, a marker molecule of hepatic stellate cell activation, was significantly increased in the model group, and the expression of α-SMA, a marker molecule of hepatic stellate cell activation, in the liver tissue of mice in the ODN A151 treatment group was significantly decreased, indicating that ODN A151 significantly inhibited the activation of hepatic stellate cells.

[0108] 7. Sirius Red staining

[0109] Sirius Red staining was performed on the liver tissue sections of the blank control group, model group, and ODN A151 treatment group to detect collagen deposition in the mouse liver tissue. The results are as follows: Fig.12 shown.

[0110] Depend on Fig.12 It can be seen that compared with the blank control group, the collagen deposition in the liver tissue of the mice in the model group was significantly increased, while the collagen deposition in the liver tissue of the mice in the ODN A151 treatment group was significantly reduced, indicating that ODN A151 significantly inhibited the formation of collagen.

[0111] The above results indicate that ODN A151 can improve liver fibrosis inflammation, reduce the occurrence of AIM2 inflammasome-dependent cell pyroptosis, reduce collagen deposition in liver tissue, reverse the symptoms of liver fibrosis in mice, and has a significant anti-fibrotic effect.

[0112] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of an AIM2 inhibitor in the preparation of a drug for treating liver fibrosis, characterized in that: The AIM2 inhibitor is ODNA151.

2. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The drug contains ODN A151 as the only active ingredient.

3. Use of the AIM2 inhibitor according to any one of claims 1 to 2 in the preparation of a drug for treating liver fibrosis, characterized in that: The medicine also includes auxiliary materials acceptable in pharmacy, food science or health care products science.

4. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The ODN A151 is used to prepare a drug for reducing the expression of liver fibrosis marker molecules COL1A1 and α-SMA and inhibiting liver fibrosis.

5. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The ODN A151 is used to prepare a drug for reducing the expression of the cell pyroptosis-related molecule GSDMD and reducing the occurrence of cell pyroptosis.

6. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The ODN A151 is used to prepare a drug for reducing the expression of pro-inflammatory cytokines IL-1β and IL-18 and improving liver fibrosis inflammation.

7. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The ODN A151 is used for preparing a medicine for reducing the degree of liver tissue structural disorder and inflammatory cell infiltration.

8. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The ODN A151 is used to prepare a lead drug for inhibiting a liver fibrosis mouse model induced by carbon tetrachloride.

9. Use of the AIM2 inhibitor according to claim 1 in the preparation of a drug for treating liver fibrosis, characterized in that: The dosage form of the drug is any one of injection, capsule, tablet, microcapsule preparation, suppository, solution, powder or spray.

10. A preparation of a drug for treating liver fibrosis, characterized in that: The therapeutically active ingredient comprises the ODN A151 described in claim 1.

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