肝龙胶囊对HBV转染的HepG2.2.15细胞的作用及其机制  

Effect of Ganlong Capsule on HBV transfected HepG2.2.15 cells and its mechanism

作　　者：刘姗 邵明园 缪志敏 黄茹润 母育成 赖泳 LIU Shan;SHAO Ming-yuan;MIAO Zhi-min;HUANG Ru-run;MU Yu-cheng;LAI Yong(College of Pharmacy and Chemistry,Dali Universitry,Dali Yunnan 611000,China)

机构地区：[1]大理大学药学与化学学院,云南大理671000

出　　处：《时珍国医国药》2020年第12期2877-2880,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81360511)。

摘　　要：目的通过体外实验观察肝龙胶囊对HBV转染HepG2.2.15细胞的抗HBV作用。方法采用CCK-8法检测肝龙胶囊对转染的HepG2.2.15细胞的半数中毒浓度(TC50)和最大无毒浓度(TC0);ELISA法检测细胞上清液中HBsAg和HBeAg的滴度;FQ-PCR法检测细胞上清液和细胞内HBV DNA含量的影响;RT-qPCR法检测细胞内STAT1、STAT2、ISGF3、STAT3、OAS、PKR、JAK2 mRNA的表达及Western-blot法检测细胞内OAS、PKR蛋白的表达。结果肝龙胶囊能有效抑制HepG2.2.15细胞HBeAg、HBsAg的分泌和HBV DNA的复制;显著增加HepG2.2.15细胞中STAT1、STAT2、ISGF3、OAS、PKR、STAT3、JAK2 mRNA和OAS、PKR蛋白的表达。结论肝龙胶囊在体外具有一定的抗HBV作用,其抗HBV的作用机制可能是通过激活JAK2-STAT3信号转导通路。Objective The anti-HBV effect of Ganlong Capsule on HBV transfected HepG2.2.15 cells was observed in vitro. Methods CCK-8 method was used to detect half therapeutic concentration(TC50) and maximum non-toxic concentration(TC0) of Ganlong Capsule on HepG2.2.15 cells;ELISA method was used to detect the titers of HBsAg and HBeAg in cell supernatant;FQ-PCR method was used to detect the effects of HBV DNA content in cell supernatant and cell;RT-q-PCR method was used to detect intracellular STAT1, STAT2, ISGF3,STAT3, OAS, PKR, JAK2 mRNA. The expressions of OAS and PKR were detected by Western-blot. Results Ganlong Capsule could effectively inhibit the secretion of HBeAg, HBsAg and HBV DNA replication in HepG2.2.15 cells, and significantly increase the expression of STAT1, STAT2, ISGF3, OAS, PKR, STAT3, JAK2, STAT3 and OAS, PKR proteins in HepG2.2.15 cells. Conclusion Ganlong Capsule has anti-HBV effect in vitro, and its anti-HBV mechanism may be through activating JAK2-STAT3 signal transduction pathway.

关 键 词：肝龙胶囊 HEPG2.2.15细胞 乙型肝炎病毒 Janus蛋白酪氨酸激酶-STAT信号转导转录活化因子 

分 类 号：R285.6[医药卫生—中药学]