Effect Of Tanshinone ⅡA Extract On Hypoxia-inducible Factor 1α And Vascular Endothelial Growth Factor In Rats With Liver Fibrosis

Objective:To observe the effect of Tanshinone II A on hypoxia-inducible factor 1α (HIF-la) and vascular endothelial growth factor(VEGF) in rats with liver fibrosis, and explore the mechanism of anti-hepatic fibrosis of Tanshinone II A.Methods:Hepatic fibrosiswas induced by intragastric carbon tetrachloride.30 male SD rats were randomly divided into five groups:carbon tetrachloride group,Tanshinone ⅡA low、middle、high(7.1、14.2、21.3mg/kg)dose groups,normally control group.Besides normally control group, the rest groups were injected with 40% carbon tetrachloride dissolved in olive oil 2.5ml/kg,twice a week, lasting 12 weeks. Normally control group was injected with same dose olive oil.After injected CCL4 2 weeks,treatment group were given 7.1、14.2、 21.3mg/kg/d Tanshinone II A by intragastric administration,carbon tetrachloride group and normally control group were given same dose of drinking water,once a day in the morning, lasting 10 weeks. After the experiment,Hydroxyproline(Hyp) contentand histopathology of liver tissue were deter-mined and the histopathology changes of liver were observed with HE staining.Immunohistochemistry method was used to observe the expression of Collagen I、TGF-β1、HIF-1α and VEGF in liver. The mRNA expressions of hepatic tissue HIF-1α、VEGF were detected with RT-PCR.Results:1、The results of the serum levels of HYP:Compared with the normally control group, the carbon tetrachloride group Hyp content are significantly increased(P<0.05).Compared with the carbon tetrachloride group,Tanshinone Ⅱ A high dose group reduced(P<0.05), and Tanshinone Ⅱ A low、middle dose groups have no statistical significance(P>0.05).2、The results of HE staining:The normally control group has no change. The hepatocyte has degenerate and necrosis,the collagen fibrils has significant hyperplasia,and false liver acinus formed in liver of the carbon tetrachloride group.The treatment effect of Tanshinone II A low dose group is not distinct,including the collagen fibrils has significant hyperplasia,false liver acinus formed in some regions. The treatment effect of Tanshinone ⅡA middle、 high dose groups are better than Tanshinone ⅡA low dose group,the structure of hepatic lobules is basic integrity, area density of collagen is increased less and there is no obvious false liver acinus formed.3、The results of immunohistochemical staining:Compared with the normally control group, the carbon tetrachloride group’s protein expressions of Col Ⅰ、TGF-β1、HIF-1α and VEGF are significantly increased(P<0.05). Compared with the carbon tetrachloride group, Tanshinone Ⅱ Alow、middle、high dose groups reduced(P<0.05).4、The results of RT-PCR:Compared with the carbon tetrachloride group, Tanshinone Ⅱ A high dose groups’ mRNA expressions of HIF-1α、 VEGF significantly reduced(P<0.05).Conclusion:1、Tanshinone Ⅱ A can reduce the content of Hyp,significantly improve the pathological change of liver tissue to reach the anti-hepatic fibrosis effect, and three dose groups have the positive relativity of dose-effect relation.2、Tanshinone ⅡA can inhibit the production of Col I, reduce the content of TGF-Pi, decreases hepatocytes apoptosis to have anti-fibrosis effects and protect the hepatocytes.3、To reduce the transcription and expression of hepatic tissue HIF-la and VEGF probably is one of Tanshinone ⅡA mechanism of anti-liver fibrosis.