Curative Effect Of AKF-PD On Rat Liver Fibrosis Induced By Pig Serum

Objective To investigate the curative effect and explore the mechanism of different doses AKF-PD on rat liver fibrosis induced by pig serum.Methods 120 male Wistar rats were randomly divided into 8 groups,the normal control group (n=15):each rat received intraperitoneal injection of 0.5ml physiological saline for twice weekly for 12 weeks; the model group(n=15) :each rat received intraperitoneal injection of 0.5ml pig serum for twice weekly for 12 weeks;four treatment groups of AKF-PD(n=15*4),IFN-a treatment group(n=15)and PFD treatment group(n=15):giving drug to all treatment groups after 8 weeks’making model,for a total of 4 weeks treatment and continue to inject pig serum.Four treatment groups of AKF-PD (60mg/kg group,120mg/kg group,240mg/kg group,480mg/kg group) and PFD treatment group,which were given corresponding doses of AKF-PD,PFD gavage once a day; IFN-a treatment group, which received subcutaneous injection of IFN-a at 10,0000 units per rat every day. At the end of 12 weekend, all the rats were killed with anaesthesia, the livers were excised for HE and Masson staining, and giving semi-quantitative scoring for inflammation and fibrosis;Immunohistochemistry was used to detect the protein expression of CollagenⅠ, CollagenⅢ;Using the Real time PCR method for CollagenⅠ, CollagenⅢ,α-SMA,TGF-β1, TIMP-1mRNA assay.Results Compared with normal group, model group’s liver tissue inflammation activity semi-quantitative score, fibrosis semi-quantitative score, collagenⅠ, collagenⅢprotein and mRNA expression were increased (p<0.001);Compared with model group, all groups of AKF-PD treatment, the inflammatory activity semi-quantitative score, fibrosis semi-quantitative score, collagenⅠ,collagenⅢprotein content, collagenⅢmRNA expression were decreased (p<0.05),collagenⅠmRNA expression of AKF-PD240mg/kg and 480mg/kg group also decreased (p<0.05);AKF-PD is better than same does of PFD on reducingⅠ,Ⅲcollagen protein content(p<0.05);AKF-PD is better than IFN-a on reducing the inflammatory activity semi-quantitative score, fibrosis semi-quantitative score, III collagen protein content, I collagen mRNA expression (p<0.05).Compared with normal group, the a-SMA, TGF-β1 and TIMP-1mRNA expression of model group were significantly higher (p<0.05);AKF-PD120mg/kg,240mg/kg and 480mg/kg group significantly reduced a-SMA mRNA expression (p<0.05),AKF-PD 240mg/kg and 480mg/kg group would decrease the expression of TGF-β1 mRNA (p<0.05),all groups could significantly reduce the amount of TIMP-1 mRNA expression (p<0.01);AKF-PD treatment groups were better than PFD treatment group and IFN-αtreatment group. Conclusion Pig serum-induced liver fibrosis in rats is successful. The AKF-PD can significantly inhibit the liver fibrosis induced by pig serum, Low-dose group (60mg/kg.d) start to effect,240mg/kg.d and 480mg/kg.d have the best effect. AKF-PD is better than IFN-a and same does of PFD on treating liver fibrosis; the anti-fibrosis effect of AKF-PD may be attributed to inhibit inflammatory reaction, decrease TGF-β1 expression, inhibit activation of HSC,reduce TIMP-1mRNA expression, promotion of extracellular matrix degradation, thus reduce theⅠ,Ⅲcollagen content in liver, to prevent liver fibrosis.