衣霉素诱导肝星状细胞内质网应激促进细胞凋亡和细胞周期阻滞  

Endoplasmic reticulum stress in hepatic stellate cells induced by tunicamycin promotes apoptosis and cell cycle arrest

作　　者：张景允 杨柳 韩萧萧 李晨驰 刘冉阳 马子华 韩冰 谢汝佳 杨勤 ZHANG Jingyun;YANG Liu;HAN Xiaoxiao;LI Chenchi;LIU Ranyang;MA Zihua;HAN Bing;XIE Rujia;YANG Qin(Guizhou Provincial Key Laboratory of Pathogensis&Drug Research for Common Chronic Diseases,Department of Pathophysiology,College of Basic Medicine,Guizhou Medical University,Guiyang 550000,China)

机构地区：[1]贵州医科大学基础医学院病理生理学教研室,贵州省常见慢性疾病发病机制及药物研究重点实验室,贵州贵阳550000

出　　处：《细胞与分子免疫学杂志》2021年第9期794-800,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81460484)。

摘　　要：目的研究衣霉素诱导HSC-T6大鼠肝星状细胞发生内质网应激(ERS)时对细胞增殖活化及细胞凋亡的影响及可能机制。方法以葡萄糖调节蛋白(GRP78)表达水平为指标摸索衣霉素诱导HSC-T6细胞ERS适宜的浓度及时间,建立衣霉素诱导HSC-T6细胞发生ERS的细胞模型。在此条件下将HSC-T6细胞随机分为对照组、1 mL/L二甲基亚砜(DMSO)处理组、1μg/mL衣霉素处理组,分别处理细胞12 h。噻唑蓝(MTT)法检测细胞增殖,流式细胞术检测细胞凋亡、细胞周期的变化;Western blot法检测α平滑肌肌动蛋白(α-SMA)、CCAAT/增强子结合蛋白子同源蛋白(CHOP)、胱天蛋白酶12(caspase-12)、细胞周期蛋白1(cyclin D1)的蛋白表达。结果筛选出衣霉素诱导HSC-T6细胞发生ERS的适宜剂量为1μg/mL,适宜时间为12 h。与对照组及DMSO处理组相比,1μg/mL衣霉素处理组细胞增殖变化不明显,但α-SMA表达上调、细胞凋亡增加,G1期细胞比例明显增加、S期细胞比例下降,ERS诱导凋亡相关信号蛋白CHOP、caspase-12均显著上调,cyclin D1表达显著下调。结论衣霉素处理HSC-T6细胞12 h可致细胞发生明显ERS并活化,活化过程中细胞数量变化不明显可能与GRP78/CHOP/caspase-12通路激活导致的细胞凋亡增多、细胞周期阻滞有关。Objective To investigate the effect of endoplasmic reticulum stress(ERS)induced by tunicamycin on proliferation,activation,and apoptosis of HSC-T6 rat hepatic stellate cells and its possible mechanism.Methods With the expression level of glucose regulated protein 78(GRP78)as an indicator to explore the optimal concentration and time,a cell model of tunicamycin-induced ERS in HSC-T6 cells was established.HSC-T6 cells were randomized into control group,treatment group with 1 mL/L of dimethyl sulfoxide(DMSO),and treatment group with 1μg/mL of tunicamycin,and the cells were treated for 12 h.MTT assay was used to detect cell proliferation,flow cytometry to detect apoptosis and cell cycle,and Western blot to detect the protein expressions ofα-smooth muscle actin(α-SMA),C/EBP cAMP homologous protein(CHOP),caspase-12,and cyclin D1.Results The optimal dose of tunicamycin to induce ERS in HSC-T6 cells was 1μg/mL and the optimal time was 12 hours.Compared with the control group and treatment group with DMSO,the treatment group with 1μg/mL of tunicamycin had no significant change in cell proliferation,but the expression ofα-SMA was up-regulated with the apoptosis increased,the proportion of G1 phase cells was significantly increased and that of S phase cells decreased,the ERS induced apoptosis related signal proteins CHOP and caspase-12 were significantly up-regulated,and the expression of cyclin D1 was significantly down-regulated.Conclusion Tunicamycin treatment of HSC-T6 cells for 12 hours induces significant ERS and activation of the cells.The insignificant change in the number of cells during the activation may be related to the increased apoptosis and the cell cycle arrest induced by the activation of the GRP78/CHOP/caspase-12 pathway.

关 键 词：肝星状细胞 衣霉素 内质网应激(ERS) 细胞增殖 细胞凋亡 

分 类 号：R965[医药卫生—药理学] R363.2[医药卫生—药学]