A small molecule compound for resisting liver fibrosis and application thereof

Abstract

The application discloses an anti-hepatic fibrosis small-molecule compound and application, which is composed of 3beta-ethoxy artenimol and 6-hydroxy kaempferol, and is used as an anti-hepatic fibrosis drug after splenectomy. The application can improve the hepatic fibrosis degree after splenectomy (reduce liver inflammation, improve liver enzyme spectrum, reduce bilirubin, and reduce collagen deposition), and the causes of liver fibrosis include at least one of alcoholic hepatitis, fatty liver, viral hepatitis, drug-induced hepatitis, autoimmune hepatitis and the like.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a small molecule compound for resisting liver fibrosis and its application. Background Art

[0002] Chronic liver disease is a major global health issue, and cirrhosis is the leading cause of death from chronic liver disease worldwide, accounting for 2.2% of global deaths in 2016. It is also the 11th leading cause of death and the 15th leading cause of morbidity worldwide. Cirrhosis is the terminal stage of liver fibrosis progression and poses a serious threat to human health, for which no satisfactory treatment is currently available. Cirrhosis and its secondary hypersplenism promote and influence each other, and the degree of liver fibrosis increases with increasing hypersplenism. Splenectomy is the mainstay of treatment; however, postoperative symptoms of spleen deficiency and blood stasis, such as jaundice, fatigue, and varices, often occur in patients, leading to liver damage and worsening liver fibrosis. Cirrhosis is the terminal stage of liver fibrosis progression, for which no ideal treatment is currently available. Splenectomy is the mainstay of treatment for patients with cirrhosis and hypersplenism and is widely used in clinical practice. However, postoperative symptoms of spleen deficiency and blood stasis, such as jaundice, fatigue, and varices, often occur in patients, leading to liver damage and worsening liver fibrosis. At present, modern medicine is still in the exploratory stage for chemical drugs to combat liver fibrosis after splenectomy, and there are no anti-liver fibrosis drugs for post-splenectomy in clinical practice.

[0003] Traditional Chinese medicine (TCM) has demonstrated unique advantages and potential in combating liver fibrosis as a key clinical treatment for liver cirrhosis. The Golden Chamber states, "Those who treat illness before it occurs should see liver disease, understand that the liver transmits it to the spleen, and should first strengthen the spleen." This laid the foundation for the later concept of "treating illness before it occurs." This concept is a key principle of TCM treatment, and the Golden Chamber emphasizes strengthening the spleen to prevent and treat liver disease. Patients undergoing splenectomy for liver cirrhosis often present with symptoms of spleen deficiency, such as fatigue, abdominal distension, poor appetite, lethargy, a plump or tender tongue, a red tongue, a thin white tongue coating, and a thready or weak pulse. Furthermore, patients may present with symptoms of blood stasis, such as a dark tongue with petechiae or ecchymosis, varicose sublingual collaterals, and a weak, delayed, or wiry pulse. This suggests that patients undergoing splenectomy for liver cirrhosis often present with symptoms of spleen deficiency and blood stasis. Therefore, clinical treatment approaches are focused on invigorating qi and strengthening the spleen, and promoting blood circulation and removing blood stasis. Atractylodes macrocephala and safflower are representative prescriptions for these functions.

[0004] 3β-Acetoxyatractylone is derived from Atractylodes macrocephala, a plant of the Asteraceae family. English: 3β-Acetoxyatractylone, CAS61206-10-8, molecular weight 274.355, C 17 H 22 O3, structural formula Figure 1 shown.

[0005] 6-Hydroxykaempferol, from the family of chrysanthemum, safflower, English: 6-Hydroxykaempferol, CAS4324-55-4, molecular weight 302.25, C 15 H 10 O7。Structural formula as Figure 2 shown.

[0006] The two substances are not used for treating liver fibrosis, and the use of the two substances for treating liver fibrosis after splenectomy can provide new ideas and directions for the development and application of drugs for liver fibrosis. SUMMARY

[0007] The purpose of the present application is to overcome the defects and deficiencies existing in the prior art, and to provide an anti-liver fibrosis small molecule compound and application, which solves the various problems existing in the prior art.

[0008] To achieve the above-mentioned purpose, the present application provides the following technical scheme:

[0009] The purpose of the present application is to provide a significant targeted therapy for liver fibrosis after splenectomy, and to provide new ideas and directions for the development and application of drugs for liver fibrosis. The small molecule compound targets AKT / NF-κB pathway, inhibits liver verification, and reduces the degree of liver fibrosis.

[0010] The purpose of the present application is to provide a significant targeted therapy for liver fibrosis after splenectomy, and to provide new ideas and directions for the development and application of drugs for liver fibrosis. The small molecule compound targets AKT / NF-κB pathway, inhibits liver verification, and reduces the degree of liver fibrosis.

[0011] An anti-liver fibrosis small molecule compound is composed of 3β-ethoxy atractylone and 6-hydroxykaempferol.

[0012] The anti-liver fibrosis small molecule compound is composed of 3β-ethoxy atractylone and 6-hydroxykaempferol in a weight ratio of 1:0.8-1.2.

[0013] The anti-liver fibrosis small molecule compound has the application of anti-liver fibrosis, and can be used as an anti-liver fibrosis drug after splenectomy.

[0014] The anti-liver fibrosis small molecule compound has the application of anti-liver fibrosis, and the dosage form of the anti-liver fibrosis drug is oral.

[0015] The anti-liver fibrosis small molecule compound has the application of anti-liver fibrosis, and the anti-liver fibrosis drug is used with a pharmaceutically acceptable carrier.

[0016] The anti-liver fibrosis small molecule compound has the application of anti-liver fibrosis, and the anti-liver fibrosis drug is used with a pharmaceutically acceptable carrier.

[0017] The application of the small molecule anti-liver fibrosis compound in anti-liver fibrosis generally requires that the pharmaceutical preparation be compatible with the mode of administration, and the pharmaceutical composition should be manufactured under aseptic conditions; the oral dosage form of the small molecule compound includes one of tablets, capsules, granules, syrups, solutions, and suspensions.

[0018] The application of the small molecule compound for resisting liver fibrosis in treating liver fibrosis comprises the following drug dosages: 10-40 mg / kg of 3β-acetyloxyatractylone and 10-40 mg / kg of 6-hydroxykaempferol.

[0019] The beneficial effects of the present invention are:

[0020] (1) The present invention can improve the degree of liver fibrosis after splenectomy (reducing liver inflammation, improving liver enzyme spectrum, reducing bilirubin, and reducing collagen deposition). The causes of liver fibrosis include at least one of alcoholic hepatitis, fatty liver, viral hepatitis, drug-induced hepatitis, and autoimmune hepatitis.

[0021] (2) This drug is easy to use, has a low preparation cost, and is easy to promote. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The chemical formula of 3β-acetyloxy atractylodesinone;

[0023] Figure 2 is the chemical formula of 6-hydroxykaempferol;

[0024] Figure 3 Drug active ingredient-disease-target network diagram;

[0025] Figure 4 Gene ontology GO enrichment histogram of intersection targets;

[0026] Figure 5 Kyoto Encyclopedia of Genes and Genomes KEGG enrichment bubble map;

[0027] Figure 6 Schematic diagram of the molecular docking of 3β-Acetoxyatractylone and CHRM1 in the simulated molecular docking of the active pharmaceutical ingredient with the key protein of the AKT / NF-κB pathway;

[0028] Figure 7 Schematic diagram of the molecular docking of 3β-Acetoxyatractylone and CHRM2 in the simulated molecular docking of the active pharmaceutical ingredient with the key protein of the AKT / NF-κB pathway;

[0029] Figure 8Molecular docking of 3β-Acetoxyatractylone and PTGS2 in the simulated molecular docking of drug active ingredients and key proteins of the AKT / NF-κB pathway;

[0030] Figure 9 Molecular docking of 3β-Acetoxyatractylone and RXRA in the simulated molecular docking of drug active ingredients and key proteins of the AKT / NF-κB pathway;

[0031] Figure 10 Molecular docking of 6-Hydroxykaempferol and HSP90AB1 in the simulated molecular docking of drug active ingredients and key proteins of the AKT / NF-κB pathway;

[0032] Figure 11 Molecular docking of 6-Hydroxykaempferol and PTGS2 in the simulated molecular docking of drug active ingredients and key proteins of the AKT / NF-κB pathway;

[0033] Figure 12 This is a graph showing the improvement of liver function indicators in cirrhotic rabbits by the compound;

[0034] Figure 13 These are pictures of the pathological changes in the liver tissue of the control group observed under an optical microscope;

[0035] Figure 14 This is a picture of the pathological changes of liver cirrhosis model tissue observed under an optical microscope;

[0036] Figure 15 These are pictures of the pathological changes of liver tissue in the splenectomy group of cirrhosis observed under an optical microscope;

[0037] Figure 16 This is a picture of the pathological changes in liver tissue in the low-dose drug group observed under an optical microscope;

[0038] Figure 17 This is a picture of the pathological changes in liver tissue of the medium-dose group observed under an optical microscope;

[0039] Figure 18 This is a picture of the pathological changes in liver tissue in the high-dose drug group observed under an optical microscope;

[0040] Figure 19 Western Blot was used to detect the expression levels of Col-I, Col-III, and α-SMA in the three groups;

[0041] Figure 20 The bar graph shows the relative expression levels of Col-I, Col-III, and α-SMA proteins in the three groups of LX-2 cells detected by Western Blot. DETAILED DESCRIPTION

[0042] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.

[0043] Embodiment 1,

[0044] A small-molecule compound for resisting liver fibrosis is composed of 3β-ethyl oxy atractylodin and 6-hydroxy kaempferol in a weight ratio of 1:1, wherein the chemical formula of 3β-ethyl oxy atractylodin is as shown in Figure 1 The chemical formula of 6-hydroxy kaempferol is as shown in Figure 2 .

[0045] The preparation method is as follows:

[0046] 30 mg of 3β-ethyl oxy atractylodin is weighed, and 30 mg of 6-hydroxy kaempferol is also weighed in a weight ratio of 1:1, and then they are dissolved in 10 ml of physiological saline to prepare a small-molecule drug combination solution.

[0047] Embodiment 2,

[0048] A small-molecule compound for resisting liver fibrosis is composed of 3β-ethyl oxy atractylodin and 6-hydroxy kaempferol in a weight ratio of 1:1.2.

[0049] 30 mg of 3β-ethyl oxy atractylodin is weighed, and 30 mg of 6-hydroxy kaempferol is also weighed in a weight ratio of 1:1.2, and 36 mg of 6-hydroxy kaempferol is also weighed in a weight ratio of 1:1.2, and then they are dissolved in 10 ml of physiological saline to prepare a small-molecule drug combination solution.

[0050] Embodiment 3,

[0051] A small-molecule compound for resisting liver fibrosis is composed of 3β-ethyl oxy atractylodin and 6-hydroxy kaempferol in a weight ratio of 1:0.8.

[0052] 30 mg of 3β-ethyl oxy atractylodin is weighed, and 24 mg of 6-hydroxy kaempferol is also weighed in a weight ratio of 1:0.8, and then they are dissolved in 10 ml of physiological saline to prepare a small-molecule drug combination solution.

[0053] The effect of the drug in Embodiment 1 of the present application is proved by experiments as follows

[0054] Experiment 1:

[0055] The physicochemical structures and targets of 3β-acetyloxyatractyl ketone and 6-hydroxykaempferol were obtained using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (http: / / tcmspw.com / tcmsp.php, TCMSP).

[0056] Disease names were calibrated using "Liver Fibrosis" on the NCBI website. The resulting names were used as keywords to search for disease targets in the Drug Target Database (TTD) (http: / / db.idrblab.net / ttd / ), the Online Mendelian Inheritance in Man (OMIM) database (https: / / omim.org / ), the GeneCards database (https: / / www.genecards.org / ), the PharmGKB database (https: / / www.pharmgkb.org / ), and the DrugBank database (https: / / go.drugbank.com / ). Targets were supplemented by reviewing relevant literature. All targets were deduplicated and gene IDs were corrected using the Uniprot database (https: / / www.uniprot.org / ) to obtain therapeutic targets corresponding to liver fibrosis (Table 1).

[0057] The common targets of 3β-acetyloxyatractylone and 6-hydroxykaempferol in liver fibrosis were observed, and the drug active ingredient-disease-target network diagram was constructed using Cytoscape 3.8.0 software (e.g. Figure 3 ).

[0058] The main targets of AKT / NF-κB pathway acted by 3β-acetyloxyatractylone and 6-hydroxykaempferol were observed, and the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of the common targets were observed (e.g. Figure 4 , Figure 5 ).

[0059] The AutoDockVina software was used to perform molecular docking of 3β-acetyloxyatractyl ketone and 6-hydroxykaempferol on CHRM1, HSP90AB1PTGS2, and RXRA targets to verify their mutual binding activity. First, the 2D structure file of the small molecule ligand was downloaded from the PubChem website and converted into a 3D structure. The pdb format file of the core target protein was downloaded from the PDB website (http: / / www.rcsb.org / ). The corresponding ligand and receptor were processed and operated using AutoDockTools. AutoDockVina was used for molecular docking. The receptor and ligand were docked with the minimum binding energy (at least less than -5kcal / mol) to obtain a molecular docking diagram (as shown in Figure 2). Figures 6-11) and the corresponding ligand-receptor binding energy (Table 1).

[0060]

[0061] Figures 6-11 This is a simulated molecular docking diagram of the active ingredient of the drug and the key protein of the AKT / NF-κB pathway.

[0062] Experiment 2

[0063] Eighty New Zealand white rabbits, 3 months old, weighing 2.5-3.0 kg and aged 90-120 days, 40 males and 40 females, were selected. The animals were housed in stacked stainless steel wire cages and fed a standard pelleted diet with free access to tap water. The rabbit rooms were well ventilated and kept clean, with an air humidity of 50%-70% and a temperature of 20°C-29°C. After acclimation for one week, blood was drawn from the central auricular artery after a 12-hour fast to assess liver function. Color Doppler ultrasonography revealed no liver or spleen lesions. Rabbits with normal indices in all groups were included in the study.

[0064] A liver cirrhosis model was prepared using the thioacetamide (TAA) method. TAA was added to normal saline to prepare a 5% solution, which was injected intraperitoneally at a dose of 30 mg / kg, twice a week, for 16 weeks. Dose adjustment plan: ① If the weight gain or loss within 1 week is within 5%, maintain the original dose. ② If the weight loss is more than 5% within 1 week, reduce the TAA dose by 20 mg / kg. ③ If the weight loss is more than 10% within 1 week, suspend the injection 1-2 times, and adjust the dose after the weight stops decreasing to avoid death. ④ If the weight increases by more than 5% continuously within 1 week, increase the TAA dose by 10 mg / kg.

[0065] 1. Splenectomy model establishment plan

[0066] Penicillin was injected one day before splenectomy, and the rabbit was fasted for 24 hours before the operation (drinking water was allowed). 20% urethane solution was injected intraperitoneally at a dose of 5 ml / kg for anesthesia. The rabbit was fixed on a fixed frame and its limbs were tied to the four corners of the frame with thin ropes. After the vital signs were stable, the rabbit's abdominal hair was shaved with pet electric clippers, and the skin was disinfected with complex iodine and alcohol. A midline incision of about 6.0 cm was made in the abdomen below the xiphoid process. The skin was cut open layer by layer to separate the subcutaneous tissue to the abdominal cavity. Sterile gauze was used to protect the gastrointestinal tract and pulled open to fully expose the visual field. After finding the spleen, the splenic artery and vein were ligated, the spleen was removed, and the abdomen was closed layer by layer after careful hemostasis. The operation was completed. The dressing was changed once a day after the operation, and a dose of 40,000 units / kg / time of penicillin was injected intramuscularly, twice a day, for 5 consecutive days.

[0067] 2 Grouping scheme

[0068] Eighty New Zealand white rabbits were randomly divided into four groups with 20 rabbits in each group using random codes generated by software: (1) blank control group; (2) liver cirrhosis group; (3) liver cirrhosis splenectomy group; (4) compound group (three subgroups): ① low-dose compound group; ② medium-dose compound group; ③ high-dose compound group.

[0069] 4 Intervention plans for each group

[0070] 4.1 Preparation of Traditional Chinese Medicine and Determination of Infusion Dosage

[0071] 4.2 Intervention plan

[0072] (1) Blank control group: No treatment was given, free access to water and food, and no saline or Chinese medicine was given.

[0073] (2) Cirrhosis group: Normal New Zealand white rabbits were injected intraperitoneally with TAA to establish a cirrhosis with hypersplenism model. The model was completed after 16 weeks. The rabbits were free to move around in the cage and had free access to water and food. They were given normal saline by gavage at 10 g / (kg.d) twice a day, at 9:00 and 15:00, for 21 consecutive days.

[0074] (3) Cirrhosis and splenectomy group: Normal New Zealand white rabbits were injected intraperitoneally with TAA to establish a cirrhosis and hypersplenism model. Splenectomy was performed 16 weeks after model establishment. The rabbits were free to move around in the cage after awakening. They were fasted on the day of surgery and allowed to drink water and eat freely from the first day after surgery. On the second day after surgery, they were given normal saline by gavage at 10 g / (kg.d) twice a day, at 9:00 and 15:00, for 21 consecutive days.

[0075] (4) Compound group

[0076] ① Low-dose group: Normal New Zealand white rabbits were injected intraperitoneally with TAA to establish a liver cirrhosis with hypersplenism model. After 16 weeks of model establishment, splenectomy was performed. The rabbits were free to move around in their cages after awakening and fasted on the day of surgery. From the first day after surgery, they were allowed to drink water and eat freely. On the second day after surgery, the rabbits were given the agent prepared in Example 1 at a dosage of 10 mg / kg 3β-acetyloxyatractylone and 10 mg / kg 6-hydroxykaempferol by gavage twice a day at 9:00 and 15:00, respectively, for 21 consecutive days.

[0077] ②Medium-dose group: Same as the low-dose group, except that the dose was changed to 20 mg / (kg.d).

[0078] ③ High-dose group: Same as the low-dose group, except that the dose was changed to 40 mg / (kg.d).

[0079] 5. Specimen collection

[0080] After the treatment, the rabbits were anesthetized and sacrificed by air embolization of the ear vein. After specimen collection, the bodies were sent to the animal center for centralized incineration. Specimen collection: (1) The cleaned, intact liver specimens were removed and collected and stored at -80°C for later use; (2) 5 ml of blood was drawn from the portal vein and inferior vena cava and stored at -80°C for later use. Sacrifice and specimen collection occurred 21 days after treatment.

[0081] 6.1.3 Blood index examination

[0082] ① Liver function indicators (alanine aminotransferase, aspartate aminotransferase, total bilirubin, direct bilirubin, indirect bilirubin, and albumin): All groups underwent fasting venous blood collection, added anticoagulants, and stored at low temperatures. Blood biochemical indicators were detected by a fully automatic biochemical analyzer (HITACHI 7600, made in Japan).

[0083] ② Detection of liver fibrosis indicators (hyaluronidase, laminin, type III procollagen, type IV collagen, and glycocholic acid): All groups underwent fasting venous blood collection, added anticoagulant, and stored at low temperature. All indicators were detected by fully automatic chemiluminescence immunoassay.

[0084]

[0085]

[0086] The low-dose compound group showed improved alanine aminotransferase, indirect bilirubin, and hyaluronidase compared with the splenectomy group for liver cirrhosis; the medium-dose compound group showed improved alanine aminotransferase, indirect bilirubin, and hyaluronidase compared with the splenectomy group for liver cirrhosis; the high-dose compound group showed improved alanine aminotransferase, alkaline phosphatase, total bilirubin, indirect bilirubin, hyaluronidase, and type III collagen compared with the splenectomy group for liver cirrhosis. The results were statistically significant (P<0.05). (As Figure 12 )

[0087] 6.1.4 Observation of liver tissue microstructure using light and electron microscopy

[0088] Liver tissue specimens were obtained from the same part, fixed with 40 g / L formaldehyde solution for 24 h, dehydrated, transparent, embedded in paraffin and sectioned. 5 μm serial sections were transparentized with xylene, dehydrated with graded ethanol, stained with hematoxylin and eosin, and mounted with neutral gum. The pathological changes of liver tissue were observed under an optical microscope.

[0089] like Figure 13-18 As shown in the figure, the liver lobule structure of the rabbits in the control group was intact, the hepatic cords were neatly arranged, the portal area was clear, the hepatic plates were distributed in a spoke-like pattern, the hepatocytes were slightly edematous, and the hepatic sinusoids were slightly dilated (red arrows) (as shown in the figure). Figure 13); In the cirrhosis model group, the liver lobule structure of rabbits disappeared, and pseudolobules of varying sizes were formed. There was inflammatory cell infiltration and bile duct cell proliferation, showing the characteristics of cirrhosis (such as Figure 14 In the splenectomy group, the normal hepatic lobule structure disappeared, the hepatocytes became edematous and balloon-like, the hepatic sinusoids expanded and became congested, and the pathological damage was alleviated (such as Figure 15 ); In the low-dose group, the normal hepatic lobule structure disappeared, the hepatic sinusoids expanded and congested, and inflammatory cell infiltration was observed (such as Figure 16 ); The liver tissue of rabbits in the medium-dose group was edematous to balloon-like degeneration, with loose cytoplasm and vacuolation, dilated hepatic sinusoids and congestion, but no other obvious abnormalities were observed (such as Figure 17 ); In the high-dose group, the rabbits had mild edema of perivenous hepatocytes, slight dilation and congestion of hepatic sinusoids, and significantly reduced pathological damage (such as Figure 18 ).

[0090] Experiment 3

[0091] Cell culture and passaging

[0092] Human hepatic stellate cell line LX-2 cells were cultured in a sterile environment at 37°C and 5% CO2 using high-glucose DMEM (complete medium) containing 10% FBS. When the cell density reached about 90%, the original medium was aspirated and the cells were washed twice with phosphate buffer. 1 mL of trypsin was added to each 90 mm culture dish and the cells were incubated in an incubator for 1-2 min. 2 mL of complete medium was added to terminate digestion. The cells were pipetted and transferred to a 15 mL centrifuge tube and centrifuged at 5000 rpm for 5 min. The upper layer of medium was aspirated and 2 mL of complete medium was added. The cells were gently pipetted and mixed. 1 mL of the medium was then transferred to a new culture dish, and the total volume of complete medium per dish was made up to 6 mL. The cells were mixed and transferred to an incubator.

[0093] Preparation of small molecule drug combination solution

[0094] Weigh 3 mg of 3β-acetyloxyatractylone and 3 mg of 6-hydroxykaempferol in a 1:1 weight ratio and dissolve in 1 ml of saline to prepare a 3 mg / mL small molecule drug combination solution. Place the prepared combination solution into a cryovial and freeze at -80°C until ready for use.

[0095] Western Blot was used to detect the effects of Chinese herbal combination solution on the expression levels of Col-I, Col-III and α-SMA

[0096] The cells were cultured in a 6-well plate. When the cell density reached 80%, the original culture medium was aspirated and the cells were washed twice with PBS. The control group was treated with complete culture medium (high-glucose DMEM containing 10% FBS), the model group was treated with complete culture medium containing 5 ng / mL TGF-β1, and the compound group was treated with complete culture medium containing 5 ng / mL TGF-β1 and 3 mg / mL small molecule drug combination liquid powder. After 48 hours of culture, the cells were digested, centrifuged, and the culture medium was removed. 100 uL SDS lysis buffer (containing 1% PMSF) was added to each well of cells, lysed on ice for 30 minutes, centrifuged, and the supernatant was collected for BCA protein quantification. 5× loading buffer was added at a ratio of 4:1, heated at 100°C for 8 minutes, and 20 μg of the denatured protein sample was subjected to gel electrophoresis, transferred to the membrane, and blocked per well. Primary antibodies (α-smooth muscle actin, α-SMA; collagen I, Col-I; collagen III, Col-III) were added and incubated overnight at 4°C. Goat anti-rabbit IgG secondary antibody was added and incubated at room temperature for 1.5 hours. After each step, the samples were washed three times with TBST for 5 minutes each time. The bands were then detected by chemiluminescence, and the grayscale values ​​of Col-I, Col-III, and α-SMA bands were analyzed using Image J. Figure 19 Western Blot was used to detect the expression levels of Col-I, Col-III, and α-SMA in the three groups; Figure 20 Western Blot was used to detect the relative expression levels of Col-I, Col-III and α-SMA proteins in the three groups of LX-2 cells.

[0097] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A small molecule compound composition for anti-liver fibrosis, characterized in that: It is composed of 3β-acetyloxy atractylodesinone and 6-hydroxykaempferol.

2. The small molecule compound composition for anti-liver fibrosis according to claim 1, characterized in that: The invention is composed of 3β-acetyloxy atractylodesone and 6-hydroxykaempferol in a weight ratio of 1:0.8-1.

2.

3. Use of the small molecule compound composition for anti-hepatic fibrosis according to claim 1 in the preparation of anti-hepatic fibrosis drugs, characterized in that: The drug is aimed at anti-liver fibrosis after splenectomy.

4. Use of the small molecule compound composition for treating liver fibrosis according to claim 3 in the preparation of an anti-liver fibrosis drug, characterized in that: The dosage form of the anti-liver fibrosis drug is an oral dosage form.

5. Use of the small molecule compound composition for preventing liver fibrosis according to claim 3 in the preparation of a drug for preventing liver fibrosis, wherein a pharmaceutically acceptable carrier is added to the drug.

6. Use of the small molecule compound composition for anti-hepatic fibrosis according to claim 5 in the preparation of anti-hepatic fibrosis drugs, characterized in that: The pharmaceutically acceptable carrier is selected from saline, buffer, glucose, water and combinations thereof.

7. Use of the small molecule compound composition for anti-hepatic fibrosis according to claim 3 in the preparation of anti-hepatic fibrosis drugs, characterized in that: The dosage form of the drug is selected from one of tablets, capsules, granules, syrups, solutions and suspensions.

Images