Pang Ran, Tao Junyan, Zhang Shuling, Zhu Jiang, Yue Xin, Zhao Lei, Ye Pian, Zhu Ying
Department of Hepatology and Infectious Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Feb;30(1):98-102. doi: 10.1007/s11596-010-0118-0. Epub 2010 Feb 14.
The anti-hepatitis B virus (HBV) effects and its mechanisms of the ethanol extracts of Hypericum perforatum L. (EHP) in vitro were explored. HepG2 2.2.15 cells, a stable HBV-producing cell line, were cultured as the model system to observe the anti-HBV effect. The viral antigens of cellular secretion, HBsAg and HBeAg, were determined by enzyme linked immunosorbent assay (ELISA). The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR. In order to understand the mechanisms of the suppression of HBV replication, all HBV promoters (Cp, Xp, S1p, S2p and Fp) with luciferase reporter gene were transfected into HepG2 cells respectively. Then the activities of viral promoters were examined by luciferase reporter assay. It was found EHP effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, as well as the extracellular HBV DNA. And EHP could selectively inhibit the activity of HBV promoter Fp. Our data suggest that EHP exerts anti-HBV effects via inhibition of HBV transcription, which helps to elucidate the mechanism underlying the potential therapeutic value of EHP.
探讨了贯叶连翘乙醇提取物（EHP）体外抗乙型肝炎病毒（HBV）的作用及其机制。以稳定产生HBV的细胞系HepG2 2.2.15细胞作为模型系统进行培养，以观察其抗HBV作用。采用酶联免疫吸附测定（ELISA）法检测细胞分泌的病毒抗原HBsAg和HBeAg。通过实时荧光定量PCR检测上清液中释放的HBV-DNA量。为了解抑制HBV复制的机制，将所有带有荧光素酶报告基因的HBV启动子（Cp、Xp、S1p、S2p和Fp）分别转染到HepG2细胞中。然后通过荧光素酶报告基因检测法检测病毒启动子的活性。结果发现，EHP能以剂量依赖的方式有效抑制HepG2 2.2.15细胞中HBsAg和HBeAg的分泌以及细胞外HBV DNA。并且EHP能选择性抑制HBV启动子Fp的活性。我们的数据表明，EHP通过抑制HBV转录发挥抗HBV作用，这有助于阐明EHP潜在治疗价值的作用机制。
