红景天苷对HepG2.2.15细胞HBV DNA复制的影响及机制研究

Effect of salidroside on HBV DNA replication of HepG2.2.15 cells and its mechanism

目的探讨红景天苷(Sal)对HepG2.2.15细胞的乙型肝炎病毒DNA(HBV DNA)、HBV前基因组RNA (pgRNA)的影响以及其可能的作用机制。方法使用0、50、100μmol/L的Sal处理HepG2.2.15细胞,分别使用CCK8、流式细胞仪、实时定量PCR检测细胞活性、细胞周期和凋亡、上清液HBV DNA、细胞内pgRNA以及相关调控基因沉默信息调节因子2相关酶1(SIRT1)、过氧化物酶体增殖物激活受体γ辅助激活因子-1α(PGC-1α)、AMP依赖蛋白激酶α2(AMPKα2)、P38蛋白激酶(P38MAPK)和Forkhead转录因子1(FOXO1)mRNA表达水平;分析pgRNA和各基因的相关性。结果 Sal对HepG2.2.15的细胞活性、细胞凋亡率、细胞周期没有较大影响;50、100μmol/L Sal处理细胞6 d后,细胞上清液中的HBV DNA较对照组明显增加(F=42.337,P=0.006);pgRNA的变化趋势与SIRT1 mRNA变化趋势的相关性明显(r=0.891),pgRNA变化趋势与其它基因PGC-1α、AMPKα2、P38MAPK、FOXO1的mRNA变化趋势相关性系数r分别为0.553、0.135、-0.943、-0.809。结论 Sal可以通过影响SIRT1途径促进HBV的复制,慢性乙型肝炎患者和HBV携带者使用Sal时应关注乙肝病毒复制情况。

Objective To explore the effects of salidroside(Sal)on hepatitis B virus DNA(HBV DNA)and HBV pregenomic RNA(pgRNA)of HepG2.2.15 cells and its possible mechanism.Methods HepG2.2.15 cells was treated with salidroside at 0,50 and 100μmol/L,respectively.Cell activity,cell cycle and apoptosis,HBV DNA in supernatant,pgRNA and the mRNA expression levels of related regulatory genes sirtuin 1(SIRT1),peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC-1α),adenosine monophosphate-activated protein kinaseα2(AMPKα2),p38 mitogen-activated protein kinase(P38 MAPK)and forkhead box protein O1(FOXO1)were detected by CCK8,flow cytometry and real-time quantitative PCR,respectively.The correlations between pgRNA and these genes was analyzed.Results The cell activity,apoptosis rate and cell cycle of HepG2.2.15 were not significantly affected by salidroside,and with 50,100μmol/L salidroside for 6 days,the HBV DNA in the supernatant was significantly higher than that in the control group(F=42.337,P=0.006).The correlation between the change trend of pgRNA and the change trend of SIRT1 mRNA was significant(r=0.891).The correlation coefficient r of the change trend of pgRNA and other genes PGC-1α,AMPKα2,P38 MAPK,FOXO1 mRNA were 0.553,0.135,-0.943 and-0.809,respectively.Conclusion Sal can promote the replication of HBV by affecting the SIRT1 pathway.In patients with chronic hepatitis B and hepatitis B virus carriers,attention should be paid to the replication of hepatitis B virus when salidroside is used.