| Objective To investigate the effect of danfang capsule on the expression of Wnt1,β-catenin and DKK1 in liver tissue of immunological liver fibrosis in rats.Methods Sixty clean grade male SD rats were divided into 6 groups on average by random number method: Model group,Dan low group,Dan medium group,Dan high group,positive control group,normal group,each group of 10.The normal group was intraperitoneally injected with the same amount of normal saline,and the other groups were intraperitoneally injected with 0.5 ml a rat pig serum to prepare rat immune liver fibrosis model,twice a week for 12 weeks.At the same time,DanFang capsules(0.054 g/100 g,0.216 g/100 g,0.432 g/100g)were given to the Dan low group,the Dan medium group and the Dan high group,respectively.The positive control group was given 0.054 g/100 g compound turtle turtle soft liver tablets,and the model group and the normal group were given the same amount of normal saline once a day for a total of 12 weeks.The rats were anesthetized and sacrificed at week 12,and the liver histopathological changes were observed by HE staining.The content of hydroxyproline(HYP)in liver tissues was detected by alkaline hydrolysis method,and the expression levels of Wnt1,β-catenin and DKK1 in liver tissues were detected by immunohistochemical method,Western Blot and Q-PCR.Results 1.Animals’ s general conditions: the rats in the whole experiment were in good condition with shiny hair.In the first week of the experiment,one died in the Dan low group and the Dan medium group due to a mistake in the operation of intragastric administration.At week 5 of the experiment,one rat in the model group died due to hemorrhagic foci in the lung,while no death occurred in the rest groups.2.Liver histopathological changes: liver fibrosis was the most serious in the model group,with extensive destruction of liver tissue parenchyma,disordered arrangement of liver cells forming a cord-like extension to the hepatic lobule,segmentation and destruction of hepatic lobule structure,and even false lobule,indicating successful modeling.Compared with the normal group,the degree of liver fibrosis was more severe in the model group,Dan low group and Dan medium group(F=53.55,42.95,39.45,all P<0.05).Compared with model group,liver fibrosis was reduced in Dan medium group and Dan high group(F=39.45,18.80,all P<0.05).Compared with the positive control group,HYP content increased in Dan low group and Dan medium group(t=8.621,-6.874,P<0.05).However,Dan high group was similar to the positive control group,and the difference was not statistically significant(P>0.05).Compared with the DanFang capsule group,the degree of liver fibrosis was reduced successively in the three groups(all P<0.05).3.HYP content in liver tissue: compared with the normal group,HYP content in the model group and Dan low group increased(t=-144.565,-28.806,all P<0.05).Compared with the model group,HYP content in the Dan medium group and Dan high group were decreased(t=92.253,165.217,all P<0.05).Compared with positive control group,the content of HYP in the Dan low adn Dan medium group increased(t=8.621,-6.874,P<0.05),but there was no significant difference between Dan high group and positive control group(P>0.05).The HYP content in the Dan low group,Dan medium group and Dan high group decreased successively,and the difference among the three groups was statistically significant(all P<0.05).3.Results of immunohistochemistry,western blot and Q-PCR in liver tissues: compared with the normal group,the expressions of Wnt1 and β-catenin were increased in the model group,Dan low group and Dan medium group(P<0.05),and the expressions of DKK1 were decreased(P<0.05).Compared with the model group,the expressions of Wnt1 and β-catenin in the Dan medium group and Dan high group were decreased(P<0.05)and the expressions of DKK1 were increased(P<0.05).Compared with the positive control group,the expressions of Wnt1,β-catenin in the Dan low group and the Dan medium group were increased(P<0.05)and the expression of DKK1 was decreased(P<0.05).There was no significant difference in Wnt1,β-catenin and DKK1 expression between Dan high group and positive control group(P>0.05).Wnt1,β-catenin gene and protein expressions decreased successively in Dan low group,Dan middle group and Dan high group,and DKK1 gene and protein expressions increased successively in them,with statistically significant differences among the groups(all P<0.05).Conclusion 1.The medium and high dose danfang capsule had the effect of intervention on rat immune liver fibrosis.There was a dose-effect relationship between different doses of danfang capsules,and the effect was better in the high-dose group.2.DanFang capsule may play an intervention role in immune liver fibrosis in rats by up-regulating the expression of DKK1 in liver tissue,reducing the expression of Wnt1 and β-catenin,and inhibiting the activation of Wnt / β-catenin pathway. |
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