Hepatitis C virus-induced hepatocyte cell death and protection by inhibition of apoptosis
Hepatitis C virus-induced hepatocyte cell death and protection by inhibition of apoptosis
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DOI：
10.1099/vir.0.065862-0
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发表时间：
2014-10-01
影响因子：
3.8
通讯作者：
Torresi, Joseph
中科院分区：
文献类型：
--
作者：
Lim, Eu Jin;El Khobar, Korri;Torresi, Joseph
慢性丙型肝炎病毒（HCV）感染导致进行性肝纤维化，导致肝硬化和肝癌。其机制尚不清楚，但肝细胞凋亡被认为起主要作用。用细胞角蛋白18（M30 CytoDEATH）和裂解的聚（ADP-核糖）聚合酶（PARK）的免疫组织化学方法测定人肝组织中的肝细胞凋亡。用表达HCV蛋白的复制缺陷型重组腺病毒（rAdHCV）进行体外研究，以研究HCV对Huh 7细胞、原代小鼠肝细胞（PMoH）和原代人肝细胞（PHH）中细胞死亡的影响。使用结晶紫测定法和蛋白质印迹法研究细胞活力和细胞凋亡，探测裂解的半胱天冬酶-3和裂解的PARP，用和不用泛半胱天冬酶抑制剂Q-VD-OPh和坏死抑素-1处理。与HCV阴性患者相比，HCV感染患者的肝组织中凋亡标志物水平升高。与未感染的对照和用对照病毒（rAdGFP）感染的细胞相比，rAdHCV感染降低细胞活力。Huh 7，PMoHs和PHHs感染rAdHCV的细胞凋亡标志物的水平显着增加与未感染的对照组和rAdGFP感染的细胞相比。在rAdHCV感染的Huh 7中，用Q-VD-OPh和necrosstatin-1处理都提高了细胞活力。Q-VD-Oph还降低了rAdHCV感染的Huh 7和PMoH中切割的PARR。已知HCV感染患者的肝脏中肝细胞凋亡增加。HCV促进原代和永生化肝细胞中的细胞死亡，并且这被Q-VD-OPh和necrostatin-1抑制。这些结果表明，HCV诱导的细胞死亡发生的细胞凋亡和坏死，并提供了新的见解HCV诱导的肝损伤的机制。
Chronic hepatitis C virus (HCV) infection results in progressive liver fibrosis leading to cirrhosis and liver cancer. The mechanism for this remains unclear but hepatocyte apoptosis is thought to play a major role. Hepatocyte apoptosis in human liver tissue was determined by immunohistochemistry for cytokeratin 18 (M30 CytoDEATH) and cleaved poly(ADP-ribose) polymerase (PARK In vitro studies were performed with replication-defective recombinant adenoviruses expressing HCV proteins (rAdHCV) to study the effects of HCV on cell death in Huh7 cells, primary mouse hepatocytes (PMoHs) and primary human hepatocytes (PHHs). Cell viability and apoptosis were studied using crystal violet assays and Western blots probed for cleaved caspase-3 and cleaved PARP, with and without treatment with the pan-caspase inhibitor Q-VD-OPh and necrostatin-1. Liver tissue of HCV-infected patients expressed elevated levels of apoptotic markers compared with HCV-negative patients. rAdHCV infection reduced cell viability compared with uninfected controls and cells infected with control virus (rAdGFP). Huh7, PMoHs and PHHs infected with rAdHCV showed significantly increased levels of apoptotic markers compared with uninfected controls and rAdGFP-infected cells. In rAdHCV-infected Huh7, treatment with Q-VD-OPh and necrostatin-1 both improved cell viability. Q-VD-Oph also reduced cleaved PARR in rAdHCV-infected Huh7 and PMoHs. Hepatocyte apoptosis is known to be increased in the livers of HCV-infected patients. HCV promoted cell death in primary and immortalized hepatocytes, and this was inhibited by Q-VD-OPh and necrostatin-1. These findings indicate that HCV-induced cell death occurs by both apoptosis and necroptosis, and provide new insights into the mechanisms of HCV-induced liver injury.