Romero Marta R, Martinez-Diez Maria C, Larena Monica G, Macias Rocio I R, Dominguez Mercedes, Garcia-Monzon Carmelo, Serrano Maria A, Marin Jose J G
Department of Biochemistry and Molecular Biology, University of Salamanca, Spain.
Antivir Chem Chemother. 2002 Nov;13(6):371-80. doi: 10.1177/095632020201300605.
A liver targeting strategy to direct antiviral drugs toward hepatitis B virus (HBV) was investigated. As model drugs we used cisplatin-bile acid derivatives (Bamets) to determine the production of virions by HBV-transfected hepatoblastoma cells (HepG2 2.2.15). Drug uptake was determined using flameless atomic absorption spectrometry to measure platinum cell contents. Cytotoxic effect was determined by formazan formation and neutral red uptake tests. The release of viral surface protein was evaluated by ELISA. The abundance of HBV-DNA in the medium was determined by quantitative real-time PCR and its structure by Southern blot analysis. The uptake of Bamets by HepG2 2.2.15 cells was higher than that of cisplatin. At concentrations lower than 10 microM, distinct Bamets have no toxic effect on host cells, whereas cisplatin dramatically reduced cell viability at concentrations higher than 1 microM. All the drugs tested inhibited the release of viral proteins to the medium, but induced a marked and progressive dose-dependent increase in the amount of viral DNA in the medium. This was mainly due to the release of short fragments of HBV-DNA in the case of cisplatin. On the contrary, Bamets induced an enhanced release of circular forms of HBV-DNA. These findings suggest the existence of a dual effect of Bamets on HBV life-cycle by enhancing the production of DNA replicative intermediates but reducing the secretion of complete virions. Altogether these characteristics recommend consideration of these compounds as a useful experimental tool in the investigation of novel liver targeted therapeutic agents based on bile acid derivatives for the treatment of HBV infections, or to carry out further studies on the HBV life cycle.
研究了一种将抗病毒药物导向乙型肝炎病毒（HBV）的肝脏靶向策略。我们使用顺铂-胆汁酸衍生物（Bamets）作为模型药物，以确定HBV转染的肝癌细胞（HepG2 2.2.15）产生病毒粒子的情况。使用无火焰原子吸收光谱法测量铂细胞含量来确定药物摄取。通过甲臜形成和中性红摄取试验确定细胞毒性作用。通过ELISA评估病毒表面蛋白的释放。通过定量实时PCR测定培养基中HBV-DNA的丰度，并通过Southern印迹分析确定其结构。HepG2 2.2.15细胞对Bamets的摄取高于顺铂。在浓度低于10 microM时，不同的Bamets对宿主细胞没有毒性作用，而顺铂在浓度高于1 microM时显著降低细胞活力。所有测试药物均抑制病毒蛋白向培养基中的释放，但诱导培养基中病毒DNA量显著且呈剂量依赖性增加。在顺铂的情况下，这主要是由于HBV-DNA短片段的释放。相反，Bamets诱导HBV-DNA环状形式的释放增加。这些发现表明Bamets对HBV生命周期具有双重作用，即增强DNA复制中间体的产生但减少完整病毒粒子的分泌。总之，这些特性建议将这些化合物作为一种有用的实验工具，用于研究基于胆汁酸衍生物的新型肝脏靶向治疗剂治疗HBV感染，或对HBV生命周期进行进一步研究。
