Agnieszka Kołakowska, Paulina Godzik, Kazimierz Madaliński
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引用次数: 0
摘要
导言:根据世卫组织的报告,全球有1.3 - 1.7亿人慢性丙型肝炎病毒感染。目前还没有针对丙型肝炎病毒的有效疫苗,目前的慢性丙型肝炎治疗标准效率有限,而且有不良的副作用。目前的研究重点是寻找一种新的治疗药物,这是专门针对该病毒。本研究的目的是开发一种方法,用于检测感染HCV的Huh-7.5细胞系中潜在解旋酶抑制剂(蒽环类抗生素的衍生物)的活性和细胞毒性。方法:采用脂质转染法感染Huh-7.5细胞系JFH1(日本暴发性肝炎)RNA。四氮唑盐XTT孵育1、2、3、4、24 h后,采用细胞增殖试剂盒(Cell Proliferation Kit II, XTT)检测蒽环类抗生素的细胞毒性。采用实时荧光定量PCR法检测蒽环类抗生素的活性。结果:优化了蒽环类抗生素的细胞毒性和活性研究条件。结论:感染HCV的Huh-7.5细胞系是筛选新的抗HCV抗病毒药物的可靠细胞培养模型。
[Methods of testing potential inhibitors of hepatitis C in Huh-7.5 cell line].
Introduction: According to WHO reports, there are 130-170 million persons chronically infected with hepatitis C virus on a global scale. There is no effective vaccine against HCV, and the current standard of chronic hepatitis C therapy has limited efficiency and undesirable side effects. Current studies are focused on searching for a new therapeutic agents, which are specifically targeted against the virus. The aim of the study was to develop a methodology for testing the activity and cytotoxicity of potential helicase inhibitors (derivatives of anthracycline antibiotics) in Huh-7.5 cell line infected with HCV.
Methods: The Huh-7.5 cell line was infected with the JFH1 (Japanese Fulminant Hepatitis) RNA by lipofection. The cytotoxicity of anthracycline antibiotics was measured by Cell Proliferation Kit II(XTT), after 1, 2, 3, 4 and 24 hours after incubation with tetrazolium salt XTT. The activity ofanthracycline antibiotics was examined by Real-Time PCR method.
Results: The study allowed to optimize the conditions of cytotoxicity and activity studies of anthracycline antibiotics.
Conclusions: Huh-7.5 cell line infected with HCV is a robust cell culture model for screening new antivirals against HCV.