Application of plant extract dauricine in prevention and treatment of acute liver injury and protection of liver

Abstract

The invention provides application of a plant extract dauricine in prevention and treatment of acute liver injury and protection of liver. Research finds that dauricine can effectively improve the liver function of an acute liver injury model mouse, reduce the liver cell damage and inflammation level, improve the oxidative stress state of liver cells and enhance the migration ability of the liver cells. Therefore, the dauricine can be used as a medicine for preventing and treating the acute liver injury and diseases related to the acute liver injury and protecting the liver, and has the advantages of definite components, controllable quality, good curative effect and the like.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to application of plant extract daemine in prevention and treatment of acute liver injury and protection of liver. BACKGROUND

[0002] Acute liver injury (ALI) is a syndrome of severe damage of hepatocytes caused by various causes in a short period of time, and the core features include massive necrosis and apoptosis of hepatocytes, severe inflammatory reaction and rapid deterioration of liver function. The clinical manifestations are jaundice, coagulopathy and significant elevation of transaminase, and severe cases can rapidly progress to acute liver failure (ALF) with high mortality.

[0003] Acute liver injury is different from liver fibrosis and liver cancer. Acute liver injury is a short-term and severe toxic injury, and the core pathological process is rapid necrosis and apoptosis of hepatocytes, accompanied by strong oxidative stress and acute inflammatory reaction. The treatment research is how to protect and repair normal liver parenchymal cells to prevent them from toxic injury, and the biological effect pursued is anti-apoptosis, anti-oxidation and anti-inflammation. Liver fibrosis is a repair reaction after long-term, chronic and continuous liver injury, and the core pathological process is activation and proliferation of hepatic stellate cells and excessive deposition of extracellular matrix (collagen, etc.). The treatment target is more focused on inhibiting activation of hepatic stellate cells, promoting apoptosis of activated hepatic stellate cells, inhibiting collagen synthesis and promoting its degradation. The treatment research of liver cancer is how to inhibit or even kill abnormally proliferating tumor cells, and the biological effect pursued is cell cycle arrest and induction of apoptosis / necrosis.

[0004] There is still a lack of effective drugs for treating acute liver injury in the art. SUMMARY

[0005] Based on this, the purpose of the present application is to provide application of plant extract daemine in prevention and treatment of acute liver injury and protection of liver.

[0006] To achieve the above-mentioned purpose, the following technical solutions are adopted in the present application.

[0007] In a first aspect, the present application provides application of daemine in preparation of a drug for preventing and treating acute liver injury.

[0008] In a second aspect, the present application provides application of daemine in preparation of a drug for preventing and treating acute liver injury related diseases.

[0009] In some embodiments, the acute liver injury related diseases include cirrhosis and liver failure.

[0010] In a third aspect, the present application provides application of daemine in preparation of a drug for protecting liver.

[0011] In some embodiments, the application comprises improving liver function.

[0012] In some embodiments, the application comprises reducing liver body index, AST level and / or ALT level.

[0013] In some embodiments, the application comprises reducing liver cell damage.

[0014] In some embodiments, the application comprises inhibiting liver cell inflammatory response.

[0015] In some embodiments, the application comprises enhancing liver cell migration ability.

[0016] In some embodiments, the application comprises improving liver cell oxidative stress state.

[0017] In some embodiments, the application comprises increasing SOD activity, increasing GSH content, reducing MDA level and / or reducing lipid peroxide level.

[0018] The present application has found that daemoropsin can effectively improve liver function of acute liver injury model mice, reduce liver cell damage and inflammation level, improve liver cell oxidative stress state, and enhance liver cell migration ability. Therefore, daemoropsin can be used as a drug for preventing and treating acute liver injury and acute liver injury related diseases, and protecting liver, and has the advantages of clear composition, controllable quality and good efficacy. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Liver phenotype of each group of mice.

[0020] Figure 2 Liver body index, serum ALT and AST level detection results of each group of mice.

[0021] Figure 3 HE staining, Sirius red staining and Masson staining results of liver tissue of each group of mice.

[0022] Figure 4 mRNA expression level detection results of related inflammatory factors of each group of mice.

[0023] Figure 5 Oxidative stress state detection results of each group of mice.

[0024] Figure 6 cck8 detection results.

[0025] Figure 7 Apoptosis detection results of each group of mice.

[0026] Figure 8 The results of the cell migration ability detection of each group. DETAILED DESCRIPTION

[0027] The experimental methods not specified in the following examples of the present application are generally carried out according to the conventional conditions, or according to the conditions recommended by the manufacturers. The various common chemical reagents used in the examples are commercially available products.

[0028] Unless otherwise defined, all technical and scientific terms used in the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0029] The terms "comprising" and "having" and any variations thereof in the present application are intended to cover the non-exclusive inclusion. For example, a process, method, device, product or equipment including a series of steps is not limited to the listed steps or modules, but optionally also includes steps not listed, or optionally also includes other steps inherent to these processes, methods, products or equipment.

[0030] In the present application, "and / or" is mentioned to describe the association relationship of the associated objects, which means that there can be three kinds of relationships, for example, A and / or B, which can represent the three cases of A alone, A and B together, and B alone.

[0031] Dauricine (DAU), CAS No. 524-17-4, molecular formula C 38 H 44 N2O6, molecular weight 624.77, is the main active ingredient derived from the traditional Chinese medicine North Radix Sophorae Tonkinensis (with the functions of clearing heat and detoxifying, dispelling wind and relieving pain).

[0032] The following is described in conjunction with the specific embodiments.

[0033] Example 1 This example studies the therapeutic effect of Dauricine on acute liver injury.

[0034] 1. Construction of acute liver injury mouse model and drug administration Model mice: SPF BALB / C mice, 6 weeks old, weighing 18-22g.

[0035] Modeling reagent: CCl4 (solvent is olive oil), the concentration of CCl4 is 5%.

[0036] Modeling method and dosage: Twenty-eight mice were randomly divided into four groups: normal control group (blank group), model group, positive control silymarin group (SLM, 50 mg / kg), and daune alkaloid group (Dau, 50 mg / kg), with seven mice in each group. Three days before modeling, mice in the positive control silymarin group, low-dose daune alkaloid group, medium-dose daune alkaloid group, and high-dose daune alkaloid group were administered daune alkaloid or silymarin via gavage three times, with each administration 24 hours apart. The model group received an equal volume of PBS buffer. Twenty-four hours after the end of administration, mice in the model group, positive control silymarin group, and daune alkaloid group were intraperitoneally injected with 200 μL of 5% CCl4 olive oil solution once, with each injection 18 hours apart. Mice in the blank group were injected with an equal volume of olive oil.

[0037] Successful modeling indicators and time points: 18 hours after modeling, the serum ALT and AST levels in the model group were detected. They were significantly higher than those in the normal control group, indicating that the modeling was successful.

[0038] 2. Indicator Testing (1) Phenotypic view Eighteen hours after the model was created, the liver was harvested for photographing.

[0039] (2) Liver body index, serum ALT and AST levels Liver function indexes were measured 18 hours after the last injection of CCl4 olive oil solution.

[0040] Eighteen hours after the last injection of CCl4 olive oil solution, serum was collected from each group of mice to detect ALT and AST levels. The method was as follows: blood was collected from the mouse eyeballs, left to stand for 6 hours, centrifuged to collect the supernatant, and sent to the testing center for testing.

[0041] (3) Liver tissue HE staining, Sirius staining, Masson staining Eighteen hours after the last injection of CCl4-olized olive oil solution, liver tissue from each group of mice was collected for HE staining, Sirius staining, and Masson staining; the methods are as follows: HE staining: 1) Fix the tissue for 48 h, then rinse with running water for 4 h. Next, soak in 75%, 85%, and 95% ethanol for 1 h each. Then soak in anhydrous ethanol I for 40 min and anhydrous ethanol II for 40 min. Then transfer to xylene I and xylene II for 20 min each. Then transfer to wax I for 40 min and wax II for 40 min (the melting point of the wax used is 56℃). Finally, embed the tissue (when performing the wax I step, the embedding machine can be opened in advance to dissolve the wax block inside the embedding machine. The melting point of the wax block used in the embedding machine is 60℃). 2) Ensure the required materials are spread evenly to avoid any cracks during the fabrication process. Next, carefully drain off excess water and gently lift it out to maintain its original shape and structural integrity. 3) First, soak in xylene I and xylene II for 10 minutes each, then transfer to anhydrous ethanol for 5 minutes. After that, transfer to 95%, 85%, and 75% ethanol for 2 minutes each, and finally soak in pure water for 2 minutes. 4) Stain the cell nuclei with hematoxylin staining solution for about 10 minutes. During this time, observe the degree of staining of the cell nuclei with a microscope. After staining, transfer the sample to water to rinse in order to remove any impurities or unwanted dye components that may remain in the staining solution. 5) After adding eosin staining solution for about 10 seconds, observe the staining effect under a microscope. Stop the staining with pure water and wash until no staining solution remains. 6) Soak in 75%, 85%, and 95% ethanol for 20 seconds each, then in anhydrous ethanol for 1 minute, and finally in xylene I and xylene II for 10 minutes each. 7) Neutral resin sealing.

[0042] Sirius staining: 1) Embedding-sectioning-dewaxing: Same as H&E staining; 2) Staining: Add Sirius red staining solution at room temperature, stain for 30 minutes, stop staining with pure water and wash until colorless; 3) Dehydration-clearing-mounting: Same as H&E staining.

[0043] Masson staining: 1) Embedding-sectioning-dewaxing: Same as H&E staining; 2) Staining: Mordant with bouin for 56°C for 1 hour, wash off bouin with water, and then stain with celestite solution for 10-15 minutes; 3) Drop Weigert iron hematoxylin staining solution (to be used on the same day) onto each tissue section and stain for 10-15 minutes; 4) Add acidic ethanol differentiation solution dropwise, 10-20 s, then wash with water; 5) Rinse the tissue sections with distilled water for 1 min; 6) Add poinsettia and magenta dye and stain for 5-10 minutes; 7) Add phosphomolybdic acid solution to induce differentiation. The differentiation time needs to be observed under a microscope until the collagen fibers turn light red and the muscle fibers turn bright red. Then wash with weak acid working solution for 1 min. 8) Shake off any remaining moisture on the slide, then add aniline blue staining solution and continue staining for 1-2 minutes; 9) After staining, rinse with a weakly acidic working solution for 1 min; 10) Dehydration-clearing-mounting: Same as H&E staining.

[0044] (4) Expression of related inflammatory factors Eighteen hours after the last injection of CCl4 olive oil solution, liver tissues of mice in each group were collected, and the mRNA expression levels of TNF-α, IL-1β, IL-6, COX2, IL-10 and GYP2E1 were detected.

[0045] The primer sequences used for detection are shown in Table 1.

[0046] Table 1 RNA extraction 1) Cut fresh liver tissue into small pieces and transfer them to homogenization tubes. Add 1 mL of Trizol and clean magnetic beads, and then homogenize by machine (homogenization conditions are the same as for protein tissue lysis). After homogenization, transfer the supernatant to an enzyme-free 1.5 mL EP tube, write the corresponding labels on the cap and tube body, and let it stand at room temperature for 5-10 minutes to allow for complete lysis. 2) Add 200 μl (1 / 5 Trizol) of chloroform, mix thoroughly by inverting, let stand at room temperature for 10 min, and then centrifuge in a low-temperature ultra-high speed centrifuge (12000 rpm, 15 min, 4℃). 3) When centrifugation is complete, carefully aspirate the clear liquid on top and transfer it to a new labeled 1.5 mL enzyme-free EP tube. Then add an equal volume of isopropanol and 1 μL of nucleic acid precipitation aid, invert and mix 10 times, and place in a 4°C refrigerator for 10 min. After centrifugation, centrifuge (12000 rpm, 10 min, 4°C). 4) Dilute anhydrous ethanol with DEPC water beforehand to prepare 75% ethanol. After centrifugation, discard the supernatant, add 1 mL of 75% ethanol to each tube, invert the tube to fully suspend the precipitate fragments in the 75% ethanol liquid, and continue centrifugation (12000 rpm, 5 min, 4℃). 5) After centrifugation, remove the supernatant, open the cap of the EP tube, and let it stand at room temperature for 5 minutes until the ethanol in the tube has completely evaporated. 6) Add 10 μL of DEPC water to each tube, mix it evenly, and determine its concentration using NanoDrop (1 μL of DEPC water is used as a blank control).

[0047] RNA reverse transcription 1) Calculate the total amount of RNA required for reverse transcription of 1 μg, add ddH2O to bring the volume to 16 μL, and then add 4 μL of 5×PrimeScript Buffer to each tube; 2) The total reverse transcription volume was 20 μL, as shown in Table 2.

[0048] Table 2 3) The reverse transcription procedure is shown in Table 3.

[0049] Table 3 Real-time quantitative PCR (qRT-PCR): 1) The total system of 20 μL = 0.4 μL rox + 10 μL mix + 0.44 μL cDNA + 8.36 μL LEPC water + 0.4 μL primer F + 0.4 μL primer R. The specific preparation system is shown in Table 4. 2) Table 5 lists the setup procedure for qRT-PCR; 3) Data usage The method is used to process and analyze the data.

[0050] Table 4 Table 5 (5) Oxidative stress state Eighteen hours after the last injection of CCl4 olive oil solution, liver tissues of mice in each group were collected to detect superoxide dismutase (SOD) activity, reduced glutathione (GSH) content, malondialdehyde (MDA) and lipid peroxide (LPO) levels.

[0051] SOD activity: Beyotime SOD kit.

[0052] GSH content: Nanjing Jiancheng GSH reagent kit.

[0053] MDA level: Nanjing has built an MDA reagent kit.

[0054] LPO level: Nanjing has built an LPO reagent kit.

[0055] (6) Bioinformatics analysis A dataset of ALI induced by carbon tetrachloride (CCl4) was obtained from the GEO database. Differentially expressed genes (DEGs) were identified using the limma package. Functional enrichment analysis was performed using the KEGG, GO, and GSEA methods. Genes were extracted from the dataset, and hub genes were identified using the MCODE and CytoHubba algorithms. Furthermore, a protein-protein interaction (PPI) network was constructed based on the String database. ImmuCellAI was used for immune cell infiltration analysis, and the Spearman method was used to determine the correlation between hub genes and immune cells.

[0056] (7) Flow cytometry Flow cytometry was used to quantitatively analyze immune cells in an acute liver injury model. The method is as follows: 1) After blood was collected from the mouse's eyeballs, the mouse was euthanized, its limbs were fixed, and its abdomen was disinfected with 75% ethanol before the abdominal cavity was opened to remove the spleen and liver; 2) Place the spleen and liver into a prepared cell strainer, add pre-cooled PBS, and grind the spleen using a grinder for 1 minute. After grinding, use a dropper to transfer the grinding buffer into a numbered 15 mL tube. Then, use a dropper to transfer the grinding buffer into the tube, rinse the cell strainer and grinder with PBS, and transfer the rinsed liquid to a 15 mL tube. Centrifuge the 15 mL tube. Centrifugation conditions: 1500 rpm, 7 min, 4 ℃; 3) After centrifugation, discard the supernatant, add 1 mL of erythrocyte lysis buffer, and gently mix with a pipette. Vortex at low speed and place on ice for 15 min, vortexing once every 5 min during this period. Add 10 mL of pre-chilled PBS to stop erythrocyte lysis, and centrifuge at 1500 rpm for 7 min at 4 °C. 4) Discard the supernatant, add 1 mL PBS to resuspend, filter through a sieve, and centrifuge again at 1500 rpm, 7 min, and 4 ℃; 5) Dispense 100 μL / tube into flow cytometry tubes, blank tubes and single staining tubes to separate the remaining cells, add 3% blocking solution to resuspend and block for 30 min. 6) Add surface staining antibody, stain at 4℃ for 30 min in the dark; 7) Add 1 mL PBS to stop staining, centrifuge at 1500 rpm for 7 min at 4℃, discard the supernatant, resuspend in 200 μL PBS, and then analyze.

[0057] (8) Immunohistochemistry Immunohistochemical staining was used to detect macrophages in an acute liver injury model. The method is as follows: 1) Same as H&E staining; 2) Add 3 g sodium citrate and 0.4 g citric acid to 1 L of pure water, then transfer to a microwave oven and cook on high for about 15 minutes. When the liquid is completely boiling, put the tissue slice into it, making sure the liquid level completely submerges the slide. Turn the microwave oven to low and continue heating for 15 minutes. After heating, take it out and let it cool naturally at room temperature for 1 hour. 3) Rinse three times with PBST (1L PBST = 1L PBS + 1 mL Tween 20), 2 min each time. Shake off the liquid on the back of the slide and around the tissue. Quickly wipe away any remaining water droplets from the tissue edges with a paper towel or cotton swab. Then, place the slide flat in a humidified chamber, select a single intact tissue, and circle it with a histochemical pen. Finally, add about 60 μl of 3% hydrogen peroxide to each circle and incubate at room temperature for 10 min (avoid light throughout the process). 4) Wash three times with PBST on a shaker at room temperature, 2 minutes each time, at a speed of approximately 70 rpm; 5) Drop approximately 60 µL of 1% BSA onto the tissue in the circle and incubate in a humidified chamber at room temperature for 1 hour; 6) Remove the blocking solution from the tissue slide. No washing is required. Place the slide in a humidified chamber and add the primary antibody (the volume should be enough to cover the tissue size). Incubate overnight at 4°C. 7) Remove the humidified chamber containing the slides from the 4°C refrigerator and quickly transfer it to a 37°C incubator for 30 min. Then, allow the chamber to cool to room temperature for 30 min. Return the chamber to the 37°C incubator and incubate for another 30 min. Next, rinse the primary antibody with PBS, followed by washing the tissue sections with PBST for 2 min each time, repeating three times. Finally, add approximately 60 μL of secondary antibody to each section. When adding the secondary antibody, carefully shake off any excess water from the slide and drop the antibody onto the tissue surface, ensuring complete coverage. Then place the slide back into the humidified chamber. Finally, incubate the chamber at room temperature for 1 h. 8) First, wash away the secondary antibody on the tissue sections with PBS, then wash three times with PBST, 2 min each time. Next is the preparation of the DAB colorimetric step. Mix the buffer and DAB concentrate thoroughly at a ratio of 20:1. Then, add approximately 60 μL of the freshly prepared DAB colorimetric solution to each tissue section, ensuring the dye completely covers the entire tissue area. To ensure optimal colorimetric development, incubate in the dark for approximately 4 min. After incubation, immediately rinse the slides with water to terminate the colorimetric reaction. 9) Stain the cell nuclei with hematoxylin for 10 min, then wash with pure water for 2 min, repeat 3 times; 10) Dehydration-clearing-mounting: Same as H&E staining.

[0058] II. Experimental Results 1. Phenotypic observation like Figure 1 As shown, compared with the blank group, the liver surface of the model group was rough and congested. After treatment with puerarin, the roughness and congestion of the liver surface were improved.

[0059] 2. Liver body index, serum ALT and AST levels like Figure 2 As shown, after CCl4 modeling, liver body index, serum ALT, and AST levels were significantly increased. After administration of silymarin and puerarin, liver body index, serum ALT, and AST levels were significantly decreased. Compared with the silymarin group, the puerarin group showed a significant decrease in ALT and AST levels. This indicates that puerarin can effectively improve CCl4-induced acute liver injury.

[0060] 3. Liver tissue HE staining, Sirius staining, Masson staining like Figure 3 As shown, HE staining results revealed no obvious degeneration, necrosis, or inflammatory infiltration in the NC group. The liver tissues of each experimental mouse exhibited different pathological changes. The CCl4 model group showed numerous necrotic foci, while the pathological changes in the silymarin and basilidine groups were improved. Sirius red staining results showed that acute CCl4 challenge induced early collagen fiber deposition in liver tissue. Basilidine treatment significantly reduced CCl4-induced collagen deposition, with a significantly reduced area of ​​collagen fiber deposition around the central vein, making it thinner and sparser. Masson staining showed a significant increase in collagen fibers in the CCl4 model group, while silymarin and basilidine administration significantly reduced collagen fibers. Furthermore, silymarin treatment significantly improved the pathological changes, collagen deposition, and increased collagen fibers caused by the CCl4 model compared to silymaridine.

[0061] 4. mRNA expression levels of relevant inflammatory factors like Figure 4 As shown, after intervention with baicalein, in acute liver injury, the DAMPs released during hepatocyte damage activate immune cells in the liver, initiating the inflammasome pathway and leading to the massive release of anti-inflammatory factors IL-6, TNF-α, and IL-1β. Under the stimulation of inflammatory signals such as TNF-α and IL-1β, the expression of Cox2 in hepatocytes is rapidly induced and significantly increased. CYP2E1 represents the initiation of acute liver injury. However, after treatment, the levels of inflammatory factors IL-6 and Cox2 significantly decreased, while IL-10, as a protective feedback mechanism against acute liver injury, rapidly increased and continued to rise in the later stages, promoting liver repair. This indicates that baicalein has a positive therapeutic effect on acute liver injury.

[0062] 5. Oxidative stress state likeFigure 5 As shown, compared with the control group, CCl4 treatment significantly reduced the activity of superoxide dismutase (SOD) and the content of reduced glutathione (GSH) in mouse liver tissue, indicating severe damage to its endogenous antioxidant defense system. Simultaneously, the levels of malondialdehyde (MDA) and lipid peroxides (LPO), key products of lipid peroxidation, significantly increased, directly confirming that CCl4 successfully induced strong oxidative stress and led to lipid peroxidation damage to hepatocyte membranes. After intervention with baicalein, the above-mentioned oxidative stress state was significantly reversed. Baicalein treatment effectively increased SOD activity and GSH content in liver tissue, restoring the body's antioxidant capacity. Correspondingly, the levels of MDA and LPO also significantly decreased compared with the model group, suggesting that lipid peroxidation damage to hepatocytes was effectively curbed.

[0063] Example 2 This embodiment investigates the effects of baicalein on apoptosis and migration of human hepatic stellate cells LX-2.

[0064] 1. Cellular experiment: CCK8 assay to detect the inhibitory effect of puerarin on LX-2 activity. LX-2 cells were treated with 0 μM, 5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 60 μM and 80 μM quinacrine for 24 h, 48 h and 72 h respectively. The effect of different concentrations of quinacrine and incubation time on the viability of LX-2 cells was detected by CCK8 (Liji Biotech kit, purchased from Bosideng).

[0065] result( Figure 6 The results showed that after 24–72 h of intervention with puerarin, the viability of LX-2 cells decreased in a dose-dependent manner. The IC50 values ​​of puerarin treatment at 24 h, 48 h, and 72 h were 22.97 µM, 28.8 µM, and 45.8 µM, respectively. Since the IC50 values ​​in the two experiments at 24 h and 48 h were relatively close, it is advisable to incubate with puerarin for 24 h in subsequent cell experiments.

[0066] 2. Effects of puerarin on LX-2 cell apoptosis LX-2 cells were divided into blank control (NC), model group, 10 μM silymarin group, 15 μM silymarin group, 25 μM silymarin group and 20 μM silymarin group. Each group treated LX-2 cells for 24 h, and then used 15 μM ccl4 to establish the cell model for 6 h. Cells were then stained with Annexin V-APC / 7-ADD double staining method to detect apoptosis.

[0067] Through flow cytometry analysis ( Figure 7It was found that 24 hours after administration, the proportion of apoptotic cells in the bat kudzu alkaloid group was significantly reduced in a dose-dependent manner, indicating that bat kudzu alkaloid alleviates apoptosis in acute liver injury.

[0068] 3. Effect of cell scratch assay on the migration ability of puerarin in LX-2 cells The effect of silymarin on the migration ability of LX-2 human hepatic stellate cells was detected by a cell scratch assay. LX-2 cells were divided into blank control (NC), 20 μM silymarin group, 30 μM silymarin group and 20 μM silymarin group. LX-2 cells in each group were treated with the drug for 24 h, followed by 6 h of modeling with 15 μM ccl4. The migration ability of LX-2 cells was then detected.

[0069] result( Figure 8 The results showed that, compared with the model group, treatment with puerarin significantly promoted the migration rate of hepatocytes. At 24 hours post-scratching, the healing rate in the model group was 35%, while the healing rate in the puerarin-treated group increased to 60%. This suggests that puerarin may play a protective and repairing role in acute liver injury by enhancing the migration capacity of hepatocytes.

[0070] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0071] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. Application of puerarin in the preparation of drugs for the prevention and treatment of acute liver injury.

2. Application of puerarin in the preparation of drugs for the prevention and treatment of acute liver injury-related diseases.

3. The application as described in claim 2, characterized in that, The acute liver injury-related diseases include cirrhosis and liver failure.

4. Application of puerarin in the preparation of hepatoprotective drugs.

5. The application as described in any one of claims 1 to 4, characterized in that, The application includes improving liver function; preferably, the application includes reducing liver body index, AST level and / or ALT level.

6. The application as described in any one of claims 1 to 4, characterized in that, The applications include reducing hepatocellular damage.

7. The application as described in any one of claims 1 to 4, characterized in that, The application includes suppressing the inflammatory response of hepatocytes.

8. The application as described in any one of claims 1 to 4, characterized in that, The applications include enhancing the migration ability of hepatocytes.

9. The application as described in any one of claims 1 to 4, characterized in that, The applications include improving the oxidative stress state of hepatocytes.

10. The application as described in claim 9, characterized in that, The applications include increasing SOD activity, increasing GSH content, reducing MDA levels, and / or reducing lipid peroxide levels.

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