白藜芦醇及其衍生物抗乙型肝炎病毒体外实验研究

 目的 探讨白藜芦醇(resveratrol,反式3,4′,5-三羟基-二苯乙烯)及其衍生物体外抗乙型肝炎病毒(HBV)的作用,及相关 的量效关系与构效关系。方法 将白藜芦醇及其衍生物作用于HepG2.2.15细胞系,MTT法检测样品对HepG2.2.15细胞的毒 性,ELISA法检测细胞上清中HBsAg,HBeAg的变化,实时荧光定量PCR法检测细胞与其上清中总HBV DNA含量,流式细胞仪 检测白藜芦醇时HepG2.2.15的凋亡作用,进行综合评价。结果 白藜芦醇对抑制HepG2.2.15细胞分泌HBsAg,HBeAg治疗指 数(TI)分别为(3.26±0.39),(2.91±0.12);在低毒或无毒浓度(0.11 mol·L^-1)下降低DNA拷贝数;呈浓度依赖性诱导 HepG2.2.15细胞凋亡。结论 白藜芦醇及其衍生物在体外有一定的抗乙型肝炎病毒作用,可能是通过抑制HBV DNA复制, 诱导感染HBV的HepG2.2.15细胞凋亡而发挥抗病毒活性。综合评价白藜芦醇及其衍生物的活性,即白藜芦醇苷(Rn1)、甲氧 基取代羟基的苷4′-甲氧基-3,3′,5-二苯乙烯-3-葡萄糖苷(Rn3)]效果较好。

Inhibitory effect of resveratrol and its derivatives on HBV in vitro

OBJECTIVE To investigate the anti-HBV effect of resveratrol and its derivatives in vitro,and observe their structure response relationship.METHODS The cytoxicity of compounds on HepG2.2.15 cell was detected using MTT.The levels of HBsAg or HBeAg in the cultured supernatant of HepG 2.2.15 were tested by ELJSA.The content of total HBV DNA was quantified by Taqman real-time PCR, and FACS was employed to test the effect of resveratrol on apoptosis in HepG2.2.15.RESULTS The therapeutic indexs(TI) of resveratrol were (3.26±0.39)(HBsAg) and (2.91±0.12)(HBeAg),respectively.Quantitative real-time-PCR result revealed that resveratrol significantly inhibited total HBV DNA copies in HepG2.2.15 cell at the concentration with lower or no cytoxicity. The effect of resveratrol on apoptosis in HepG2.2.15 appeared aconcentration-dependent pattern.CONCLUSION Resveratrol is effective in inhibiting HBV in vitro, perhaps by inhibiting HBV copy and inducing apoptosis in HepG2.2.15 cell. In general, the inhibitory effect on HBsAg, HBeAg and HBV DNA copy number in vitro shows that resveratrol-glycoside and 4'-methoxy-3,3,5'-stilbene-3-glucoside are more effective than resveratrol.