中国病理生理杂志2017，Vol.33Issue(5)：913-918,6.DOI:10.3969/j.issn.1000-4718.2017.05.024
辛二酰苯胺异羟肟酸诱导大鼠原代肝星状细胞凋亡
Suberoylanilide hydroxamic acid induces apoptosis of rat hepatic stellate cells in vitro
摘要
Abstract
AIM:To study the effects of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hepatic stellate cells (HSCs) and expression of associated proteins, and to investigate the mechanisms of SAHA to induce apoptosis.METHODS:The rat HSCs were isolated by OptiPrep gradient centrifugation method.The effect of SAHA on HSC proliferation was detected by real-time cell analyzer.The morphological changes of HSCs treated with SAHA at different concentrations were observed under inverted microscope.The apoptotic rates of HSCs were analyzed by flow cytometry with Annexin V-FITC/PI staining and fluorescence microscopy.The protein expression of α-smooth muscle actin (α-SMA), collagen I, tissue inhibitor of metalloproteinase 1 (TIMP1), glucose-regulated protein 78 (GRP78) and histone deacetylase 6 (HDAC6) was detected by Western blotting.The interaction of GRP78 with HDAC6 in the HSCs was determined by co-immunoprecipitation.RESULTS:HSCs were successfully isolated and cultured for 14 d, during which the HSCs changed gradually from rest state to active state.SAHA significantly inhibited the proliferation of HSCs in a time-and dose-dependent manner (P<0.05).The results of Western blotting showed that the protein expression levels of α-SMA, TIMP1, collagen-I and HDAC6 were significantly decreased (P<0.05), while GRP78 was significantly increased (P<0.05).Compared with activated HSCs, GRP78 and total acetyl-lysine protein were significantly increased in the co-immunoprecipitated HSCs treated with SAHA, while HDAC6 protein was significantly decreased, indicting that GRP78 formed a complex with HDAC6.CONCLUSION:The anti-hepatic fibrosis effect of SAHA may be related to down-regulation of HDAC6 and up-regulation of acetylated GRP78, thus inducing endoplasmic reticulum stress of HSCs and promoting the apoptosis of HSCs.关键词
肝星状细胞/辛二酰苯胺异羟肟酸/肝纤维化/细胞凋亡Key words
Hepatic stellate cells/Suberoylanilide hydroxamic acid/Hepatic fibrosis/Apoptosis分类
医药卫生引用本文复制引用
刘幸,田甜,余蕾,詹玮,韩冰,谢汝佳,杨婷,罗新华,杨勤..辛二酰苯胺异羟肟酸诱导大鼠原代肝星状细胞凋亡[J].中国病理生理杂志,2017,33(5):913-918,6.基金项目
国家自然科学基金资助项目(No.81160064) （No.81160064）
省卫计委基金资助项目(No.gzwjkj2015-1-037) （No.gzwjkj2015-1-037）
