Thromboxane A2 Synthase Inhibitors Prevent Production of Infectious Hepatitis C Virus in Mice with Humanized Livers.
Thromboxane A2 Synthase Inhibitors Prevent Production of Infectious Hepatitis C Virus in Mice with Humanized Livers.
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血栓素 A2 合酶抑制剂可防止具有人源化肝脏的小鼠产生传染性丙型肝炎病毒。
DOI：
10.1053/j.gastro.2013.05.014
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发表时间：
2013
期刊：
影响因子：
29.4
通讯作者：
Hijikata M.
中科院分区：
文献类型：
--
作者：
Abe Y;Aly HH;Hiraga N;Imamura M;Wakita T;Shimotohno K;Chavama K;Hijikata M.
背景与目的永生化人类肝细胞（HuS-E/2 细胞）的 3 维 (3D) 培养系统最近被证明可以支持血源性丙型肝炎病毒 (HCV) 的生命周期。我们使用该系统来识别在 3D 培养条件下 HCV 生命周期中活跃的蛋白质。方法我们比较了在 2 维和 3D 条件下培养的 HuS-E/2 细胞的基因表达谱。我们确定了细胞中差异激活的信号通路，并使用重组 HCV 产生细胞培养系统以及小干扰 RNA 和化学试剂分析了它们在 HCV 生命周期中的功能。我们研究了抗 HCV 试剂在人源化肝脏（携带人类肝细胞）的小鼠中改变这些信号通路的效果。结果微阵列分析显示，在 2 维与 3D 条件下培养的细胞表达不同水平的编码前列腺素合酶的信使 RNA。小干扰 RNA 介导的血栓素 A2 合酶 (TXAS) 敲低以及肝细胞与 TXAS 抑制剂的孵育表明，这种酶是产生感染性 HCV 所必需的，但不会影响 HCV 基因组的复制或颗粒释放。 TXAS抑制剂和前列腺素I2受体激动剂具有与血栓素A2相反的作用，可降低HCV血清水平并抑制小鼠血源性HCV对人肝细胞的感染。结论前列腺素合酶抑制剂TXAS可抑制培养肝细胞中感染性HCV颗粒的产生 以及人源化肝脏小鼠肝细胞的 HCV 感染。因此，它可能对 HCV 感染有治疗作用。
Background & AimsA 3-dimensional (3D) culture system for immortalized human hepatocytes (HuS-E/2 cells) recently was shown to support the lifecycle of blood-borne hepatitis C virus (HCV). We used this system to identify proteins that are active during the HCV lifecycle under 3D culture conditions.MethodsWe compared gene expression profiles of HuS-E/2 cells cultured under 2-dimensional and 3D conditions. We identified signaling pathways that were activated differentially in the cells, and analyzed their functions in the HCV lifecycle using a recombinant HCV-producing cell-culture system, with small interfering RNAs and chemical reagents. We investigated the effects of anti-HCV reagents that altered these signaling pathways in mice with humanized livers (carrying human hepatocytes).ResultsMicroarray analysis showed that cells cultured under 2-dimensional vs 3D conditions expressed different levels of messenger RNAs encoding prostaglandin synthases. Small interfering RNA–mediated knockdown of thromboxane A2 synthase (TXAS) and incubation of hepatocytes with a TXAS inhibitor showed that this enzyme is required for production of infectious HCV, but does not affect replication of the HCV genome or particle release. The TXAS inhibitor and a prostaglandin I2 receptor agonist, which has effects that are opposite those of thromboxane A2, reduced serum levels of HCV and inhibited the infection of human hepatocytes by blood-borne HCV in mice.ConclusionsAn inhibitor of the prostaglandin synthase TXAS inhibits production of infectious HCV particles in cultured hepatocytes and HCV infection of hepatocytes in mice with humanized livers. It therefore might be therapeutic for HCV infection.