肝康颗粒的药效学研究

Pharmacodynamics research of Gankang granule

目的该实验旨在观察中药复方剂肝康颗粒体外抗乙型肝炎病毒及其药效学。方法利用HepG2.2.15细胞模型,采用MTT法和细胞凋亡染色实验来检测肝康颗粒的细胞毒性,并观察其量-效关系;通过酶联免疫吸附测定法(ELISA)检测乙肝病毒HBV中HBsAg和HBeAg的表达,并用荧光定量PCR技术定量分析HBV DNA的复制浓度。结果 MTT法检测结果发现随着浓度和时间的增加,对HepG 2.2.15细胞生长抑制作用明显增强。ELISA法测定加药后HBV病毒分泌到上清液中的HBsAg和HBeAg含量,可见随着浓度的增大,对HBsAg和HBeAg分泌的抑制作用增加,并且抑制效果随着作用时间的累加也相应增强。GKKL抑制HBV病毒分泌HBsAg和HBeAg的治疗指数值TI均大于2,结果显示药物低毒高效,具有很好的安全性。实时荧光定量PCR法检测发现,与对照组相比较,肝康颗粒对HBV DNA具有显著的抑制作用,均可使细胞上清液中HBV DNA的拷贝数降低(P<0.05,P<0.01)。且随着浓度和作用时间的增加其作用逐渐增强,呈现一个明显的量效和时效反应关系。结论由此证明肝康颗粒具有降低HBsAg和HBeAg,抑制HBV DNA复制的明显作用,将为进一步药理毒理研究提供实验基础。

【Objective】 This experiment is designed to observe the antiviral action in vitro and immune adjustment effect of traditional Chinese medicine Gan Kang Ke Li（GKKL） in treating chronic viral hepatitis B induced by hepatitis B virus（HBV）.【Methods】 Using HepG 2.2.15 cell model,MTT experiment and apoptosis dyeing experiments were made to detect cytotoxicity of GKKL and observe the dose-effect relationship.Through enzyme-linked immunosorbent assay（ELISA）,the expression level of HBeAg and HBsAg by HBV was detected.Using fluorescence quantitative PCR technology,quantitative analysis of replication concentration of HBV DNA was determined.【Results】 The MTT and ELISA method test results found that the GKKL could inhibit the growth of HepG 2.2.15 cell line and reduce the expression level of HBeAg and HBsAg along with the increase of drug concentrations.On days 3,6,9,the therapy indexes（TI） of GKKL were all larger than 2,which showed that this compound is low-toxic,secure and effective.Time fluorescence quantitative PCR method tested found that,compared with control group,GKKL has a significant inhibitory effect of HBV DNA（P 0.05,P 0.01）.And with the increase of concentration and action time,its effect presents a clear dosage-effect and time-effect relationship.【Conclusions】 It says that GKKL plays an important role in reducing HBeAg and HBsAg,inhibiting HBV DNA replication and enhancing the mice immune function.This provides a solid basis for our further exploration on this new compound.