Fluoroquinolones inhibit HCV by targeting its helicase
Fluoroquinolones inhibit HCV by targeting its helicase
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DOI：
10.3851/imp1937
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发表时间：
2012-01-01
影响因子：
1.2
通讯作者：
Ali, Syed H.
中科院分区：
文献类型：
--
作者：
Khan, Irfan A.;Siddiqui, Sammer;Ali, Syed H.
背景：全世界HCV感染者超过1.7亿。针对HCV的有效治疗仍然缺乏，并且需要开发针对该病毒的强效药物。在本研究中，我们采用了两种培养模型来测试氟喹诺酮类药物对HCV的活性：能够在细胞系Huh-8和Huh-7细胞培养模型中独立复制的亚基因组复制子，该模型采用合成HCV RNA转染的细胞来产生感染性HCV颗粒。氟喹诺酮类药物也显示出对某些病毒具有抑制活性，可能是通过靶向病毒解旋酶。方法：将12种不同的氟喹诺酮类药物分别加入到产生HCV病毒粒子的Huh-7细胞和Huh-8细胞中，观察它们对HCV NS 3解旋酶蛋白的影响。然后，裂解Huh-7和Huh-8细胞并提取病毒RNA。提取的RNA进行逆转录，并通过实时定量PCR进行定量。氟喹诺酮类药物也测试纯化的NS 3蛋白在分子信标为基础的体外helicaseassay.Results：在不同程度上，所有测试的氟喹诺酮类药物有效地抑制HCV复制在Huh-7和Huh-8培养模型。12种氟喹诺酮类药物均能抑制HCV NS 3解旋酶活性。结论：氟喹诺酮类药物可能通过靶向HCV NS 3解旋酶抑制HCV复制。这些药物有望用于治疗HCV感染。
Background: HCV has infected >170 million individuals worldwide. Effective therapy against HCV is still lacking and there is a need to develop potent drugs against the virus. In the present study, we have employed two culture models to test the activity of fluoroquinolone drugs against HCV: a subgenomic replicon that is able to replicate independently in the cell line Huh-8 and the Huh-7 cell culture model that employs cells transfected with synthetic HCV RNA to produce the infectious HCV particles. Fluoroquinolones have also been shown to have inhibitory activity against certain viruses, possibly by targeting the viral helicase. To tease out the mechanism of the antiviral activity of fluoroquinolones, their effect on HCV NS3 helicase protein was also tested.Methods: Huh-7 cells producing the HCV virion as well as Huh-8 cells were grown in the presence or absence of 12 different fluoroquinolones. Afterwards, Huh-7 and Huh-8 cells were lysed and viral RNA was extracted. The extracted RNA was reverse transcribed and quantified by real-time quantitative PCR. Fluoroquinolones were also tested on purified NS3 protein in a molecular-beacon-based in vitro helicase assay.Results: To varying degrees, all of the tested fluoroquinolones effectively inhibited HCV replication in both Huh-7 and Huh-8 culture models. The inhibition of HCV NS3 helicase activity was also observed with all 12 of the fluoroquinolones.Conclusions: Fluoroquinolones inhibit HCV replication possibly by targeting the HCV NS3 helicase. These drugs hold promise for the treatment of HCV infection.