Newcastle disease virus represses the activation of human hepatic stellate cells and reverses the development of hepatic fibrosis in mice
Newcastle disease virus represses the activation of human hepatic stellate cells and reverses the development of hepatic fibrosis in mice
复制标题
新城疫病毒抑制人肝星状细胞的活化并逆转小鼠肝纤维化的发展
DOI：
10.1111/j.1478-3231.2009.01971.x
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发表时间：
2009-04-01
影响因子：
6.7
通讯作者：
Bian, Huijie
中科院分区：
文献类型：
--
作者：
Li, Ya-Lin;Wu, Jiao;Bian, Huijie
活化的肝星状细胞（HSC）是导致肝纤维化的关键因素，并通过与肿瘤细胞相互作用参与肝细胞癌（HCC）的发展。新城疫病毒（NDV）具有在瘤形成细胞和转化细胞中本质选择性复制的溶瘤特性。但是，NDV 在 HSC 中的复制以及对肝纤维化的影响尚未有报道。我们通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定和逆转录酶聚合酶链反应 (RT-PCR) 检测了人 HCC 细胞的条件培养基 (CM) 对人 HSC 系 LX-2 激活的影响。通过流式细胞术或荧光显微镜评估NDV在LX-2细胞和原代培养的小鼠HSC中的复制。 NDV 感染后，通过明胶酶谱、RT-PCR、蛋白质印迹和天狼星红染色测定 HSC 和肝纤维化小鼠模型的肝纤维化指数。通过双重免疫荧光染色检测NDV病毒体和α-平滑肌肌动蛋白（α-SMA）的共定位。采用TdT介导的dUTP缺口末端标记法检测NDV处理小鼠肝组织中的细胞凋亡。肿瘤-CM和转化生长因子-β1（TGF-β1）可以促进LX-2细胞的增殖和活化，表现为α-SMA、胶原蛋白I、金属蛋白酶组织抑制剂表达增强。 (TIMP)-1和TGF-β1。激活的HSCs促进NDV的复制，从而抑制MMP的分泌，抑制这些肝纤维化指标的表达以及四氯化碳诱导的小鼠肝纤维化中α-SMA和胶原纤维的表达。HCC细胞促进HSCs的激活，NDV减弱 通过在活化的 HSC 中选择性复制来激活并抑制肝纤维化。
Activated hepatic stellate cells (HSCs) are the crucial factor responsible for liver fibrosis and involved in development of hepatocellular carcinoma (HCC) by interaction with tumour cells. Newcastle disease virus (NDV) has the oncolytic characteristics of intrinsically selective replication in neoplasia cells and transformed cells. But, NDV replication in HSCs and effects on hepatic fibrosis have not been reported.We detected the effect of conditioned medium (CM) from human HCC cells on the activation of human HSC line, LX-2 by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and reverse transcriptase-polymerase chain reaction (RT-PCR). The replication of NDV was evaluated in LX-2 cells and primary-cultured mouse HSCs by flow cytometry or by a fluorescence microscope. Indices for hepatic fibrosis were determined in HSCs and a hepatic fibrosis mouse model by gelatin zymography, RT-PCR, Western blot and Sirius red staining after NDV infection. Colocalization of NDV virions and alpha-smooth muscle actin (alpha-SMA) were detected by double immunofluorescence staining. Detection of apoptosis was carried out in liver tissues of NDV-treated mice by the TdT-mediated dUTP nick-end labelling assay.Tumour-CM and transforming growth factor-beta 1 (TGF-beta 1) could promote the proliferation and activation of LX-2 cells, indicated by the enhanced expression of alpha-SMA, collagen I, tissue inhibitor of metalloproteinase (TIMP)-1 and TGF-beta 1. Activated HSCs facilitated the replication of NDV, thereby repressing the secretion of MMP, the expression of these indices for hepatic fibrosis and the expression of alpha-SMA and collagen fibrils in hepatic fibrosis of the mouse induced by carbon tetrachloride.HCC cells promote the activation of HSCs and NDV attenuates the activation and represses the hepatic fibrosis by selective replication in activated HSCs.