白藜芦醇抗乙型肝炎病毒及肝炎所致纤维化的初步研究

Preliminary study of resveratrol on anti-hepatitis B virus and hepatitis induced fibrosis

目的初步研究白藜芦醇抗乙型肝炎病毒(hepatitis B virus,HBV)及肝纤维化的作用。方法采用r AAV8-1.3HBV腹腔注射新生SD大鼠建立大鼠HBV感染模型,建模成功后将大鼠随机分为4组,每组5例,即阴性对照组、白藜芦醇低剂量组、白藜芦醇高剂量组、阳性对照组,分别给予生理盐水、20 mg/kg白藜芦醇、40 mg/kg白藜芦醇、0.1 mg/kg阿昔洛韦灌胃。2、4、8、12周后尾静脉采血检测乙肝表面抗原(HBs Ag)、乙型肝炎E抗原(HBe Ag)、乙肝病毒的脱氧核糖核酸(HBV-DNA)。建立HSC-T6细胞模型,分别以生理盐水、白藜芦醇灌胃大鼠制备对照组及白藜芦醇血清,采用Alamar Blue法观察白藜芦醇血清对HSC的增殖作用,Realtime PCR法分析白藜芦醇血清对HSC细胞I型胶原及TGF-β1 mRNA表达的影响。Western Blot法分析白藜芦醇血清对HSC细胞I型胶原及TGF-β1蛋白表达的影响。结果采用ELISA法检测HBs Ag、HBe Ag、HBV-DNA发现第2周、第4周阴性对照组HBs Ag、HBe Ag的表达明显高于其余3组P<0.05,其中第2周对照组HBs Ag、HBe Ag的表达最高分别为(4118±367)IU/m L、(160.2±56.90)IU/m L。而HBV-DNA则在第2周阴性对照组与其余3组相比具有明显差异P<0.05。Alamar Blue法结果白藜芦醇血清可抑制HSC-T6增殖,Realtime PCR结果白藜芦醇血清可以显著抑制HSC-T6的I型胶原及TGF-β1 mRNA表达。Western blot结果显示白藜芦醇血清可以显著抑制HSC-T6的I型胶原及TGF-β1蛋白表达。结论白藜芦醇能抑制HBV同时通过抑制I型胶原及TGF-β1达到阻止肝纤维化的结果。

Objective To study the effect of resveratrol on hepatitis B virus（ HBV） and hepatic fibrosis. Methods The rat model of HBV infection was established by intraperitoneal injection of r AAV8-1. 3HBV. After successfully establishing the modle,were randomly divided into four groups：negative control group,resveratrol low dose group,high dose group,positive control group,and five rats in each group. Giving a physiological saline,20 mg / kg respectively resveratrol,40 mg / kg,0. 1 mg / kg acyclovir via gastrogavage respectively. 2、4、8、12 weeks later,we detected the HBs Ag,HBe Ag,HBV-DNA via tail vein blooding. HSC-T6 cell model was established. Then rats were divided into two groups,control and resveratrol serum group. Each group was administrated with normal sodium and resveratrol via gastrogavage to make serum. The HSC-T6 viability was observed by Alamar Blue assay. The expression of Col I and TGP-β1 mRNA was determined by Realtime PCR. The expressions of Col I and TGP-β1 Protein was analyzed by Western Blot. Results Using ELISA method to detect HBs Ag、HBe Ag、HBV-DNA and found that the expression of HBs Ag and HBe Ag of negative control group were significantly higher than the rest of the three groups（ P〈0. 05） at the second and the fourth week. The highest expression of HBs Ag and HBe Ag were（ 4118 ± 367） IU / m L、（ 160. 2 ± 56. 90） NCU / m L at the second week. The expression of HBV-DNA of negative control group is significantly higher than the rest of the three groups（ P〈0. 05） at the second week. Alamar Blue assay indicated that different concentration of serum of resveratrol can inhibit HSC-T6 proliferation. Compared with control group,serum with drug resveratrol significantly down-regulated the mRNA and protein expression of Col I and TGP-β1. Conclusion Resveratrol inhibits HBV and liver fibrosis by inhibiting type I collagen and TGF-β1.