Depletion of Thymosin β4 Promotes the Proliferation, Migration, and Activation of Human Hepatic Stellate Cells
Depletion of Thymosin β4 Promotes the Proliferation, Migration, and Activation of Human Hepatic Stellate Cells
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DOI：
10.1159/000363005
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发表时间：
2014-01-01
影响因子：
--
通讯作者：
Chen, Yingwei
中科院分区：
文献类型：
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作者：
Xiao, Yongtao;Qu, Chunying;Chen, Yingwei
背景和目标：最近有报道称，胸腺素β-4（T β 4）在体外对人肝星状细胞（HSC）具有抗纤维化作用，但这些作用的机制尚不清楚。本研究旨在探讨T β 4在HSC增殖、迁移和活化中的作用。研究方法：酶联免疫吸附试验（ELISA），免疫组织化学和蛋白质印迹法用于确定T β 4在血清，肝组织和LX-2细胞中的表达水平。使用小干扰RNA（siRNA）在LX-2细胞中耗尽T β 4。使用细胞计数试剂盒-8（CCK-8）活力测定分析细胞增殖，并使用伤口愈合和transwell迁移测定研究细胞迁移。结果：T β 4在肝纤维化过程中表达明显降低。T β 4的耗竭通过激活PI 3 K/Akt信号通路促进LX-2细胞的增殖和迁移。LX-2细胞中T β 4耗竭的促迁移和促增殖作用可通过Akt抑制剂MK-2206治疗来抵消。此外，T β 4耗竭还通过α-平滑肌肌动蛋白（α-SMA）和波形蛋白的表达增强与HSC活化相关。结论：我们的研究结果表明，T β 4通过抑制HSC的迁移、增殖和活化而参与肝纤维化，并且T β 4可能是治疗肝纤维化的有效靶点。版权所有（C）2014 S. Karger AG，巴塞尔
Background & Aims: It has recently been reported that thymosin beta-4 (T beta 4) has anti-fibrogenic effects in human hepatic stellate cells (HSCs) in vitro, but the mechanisms underlying these effects remain unclear. The aim of this study was to investigate the roles of T beta 4 in the proliferation, migration, and activation of HSCs. Methods: Enzyme linked immunosorbent assays (ELISA), immunohistochemistry, and western blot assays were utilized to determine the expression levels of T beta 4 in serum, liver tissues, and LX-2 cells. T beta 4 was depleted in LX-2 cells using small interfering RNAs (siRNAs). Cell proliferation was analyzed using cell counting kit-8 (CCK-8) viability assays, and cell migration was investigated using wound healing and transwell migration assays. Results: The expression of T beta 4 was significantly reduced during the progression of liver fibrosis. The depletion of T beta 4 significantly promoted the proliferation and migration of LX-2 cells via the activation of the PI3K/Akt signaling pathway. The pro migratory and pro proliferative effects of T beta 4 depletion in LX-2 cells can be counteracted by treatment with the Akt inhibitor MK-2206. In addition, T beta 4 depletion was also associated with the activation of HSCs via the enhanced expression of alpha-smooth muscle actin (alpha-SMA) and vimentin. Conclusions: Our results suggest that T beta 4 participates in liver fibrosis by inhibiting the migration, proliferation, and activation of HSCs and that T beta 4 may be an effective target in the treatment of liver fibrosis. Copyright (C) 2014 S. Karger AG, Basel