葫芦茶苷调控JAK/STAT信号通路抗乙肝病毒作用及其机制研究  

Study on anti-hepatitis B virus activities and mechanisms of tadehaginoside by regulating JAK/STAT signaling pathway

作　　者：唐爱存[1] 王明刚[1] 卢秋玉[2] 王兵[1] 苏金妹 

机构地区：[1]广西中医药大学第一附属医院,南宁530023 [2]广西壮族自治区人民医院,南宁530021 [3]广西国际壮医医院,南宁530023

出　　处：《中药药理与临床》2017年第1期74-77,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：广西自然基金项目(2015GXNSFBA139126);广西壮族自治区卫生计生委研究项目(No.Z2015481)

摘　　要：目的:探讨葫芦茶苷体外抗乙型肝炎病毒的作用以及对JAK/STAT信号通路的调控作用机制。方法:以HBV基因转染的HepG2.2.15体外细胞模型,采用MTT法检测葫芦茶苷对HBV基因转染的HepG2.2.15细胞的半数毒性浓度(TC50)和最大无毒浓度(TC0),在最大无毒浓度(TC0)情况下观察不同浓度药物作用于HepG2.2.15细胞后,分别在第4 d和8 d收集细胞培养上清液,采用ELISA法测定上清液HBsAg,HBe Ag的滴度。FQ-PCR法检测上清液HBV DNA的含量;采用FQ-PCR法检测葫芦茶苷10μg/ml,20μg/ml,40μg/ml剂量组对HepG2.2.15细胞内信号转导和转录活化因子STAT1mRNA、STAT2mRNA表达的影响;RTPCR法检测JAK2、STAT3mRNA表达水平。结果:葫芦茶苷对HepG2.2.15细胞毒性较低,TC50为109.56μg/ml,TC0为41.54μg/ml。与空白对照组比较,10μg/ml,20μg/ml,40μg/ml剂量的葫芦茶苷能有效抑制HepG2.2.15细胞分泌HBsAg和HBe Ag,明显降低HBV DNA的含量。10μg/ml,20μg/ml,40μg/ml剂量的葫芦茶苷能明显升高细胞内信号转导和转录活化因子STAT1、STAT2、STAT3、JAK2 mRNA的水平。结论:葫芦茶苷体外具有显著的抗乙型肝炎病毒作用,其机制可能是通过激活JAK/STAT信号转导通路而发挥抗病毒作用。Objective： To investigate anti-hepatitis B virus effects of tadehaginoside（ TA） in vitro,and expound regulatory mechanisms of TA on the JAK/STAT signaling pathway.Methods： The HepG2.2.15 cells transfected with HBV gene were chosen as the cell model.The TC50 and TC0of tadehaginoside（ TA） to the HepG2.2.15 cells transfected with HBV gene were determined by MTT assay.At non-cellulotoxic concentration,the 10μg/ml,20μg/ml,40μg/ml concentrations of TA were added respectively to Hep G2.2.15 cells,after 4 d’ and 8 d’ incubation,the culture medium was collected for determining the quantity of HBV-DNA by FQ-PCR and the levels of HBs Ag and HBe Ag by ELISA.FQ-PCR method was also used to detect the expression of STAT1 mRNA and STAT2 mRNA in the HepG2.2.15 cells,and RT-PCR method was also used to detect the expression of STAT3 mRNA and JAK2 mRNA.Results： Research findings show that the tadehaginoside（ TA） had little cellulotoxicity,TC50= 109.56μg/ml,and TC0= 41.54μg/ml.Compared with the control group,TA at 10μg/ml,20μg/ml,40μg/ml concentration could suppress HBs Ag and HBe Ag expressions and decrease the HBV-DNA level in HepG2.2.15 cell line（ P 〈 0.05,P 〈 0.01）,and the levels of STAT1,STAT2,STAT3 and JAK2 mRNA in Hep G2.2.15 cell were obviously increased by tadehaginoside（ 10μg/ml,20μg/ml,40μg/ml）.Conclusion： Tadehaginoside could inhibit significantly HBV in vitro,the mechanism may be related with activating JAK/STAT signal transduction pathway.

关 键 词：葫芦茶苷 乙型肝炎病毒 HEPG2.2.15细胞 JAK/STAT信号通路 

分 类 号：R285[医药卫生—中药学]