淫羊藿苷通过抑制内质网应激减轻异丙肾上腺素诱导的H9c2细胞肥大损伤  

Icariin reduces isoproterenol-induced H9c2 cell hypertrophy injury by inhibiting endoplasmic reticulum stress

作　　者：周琳 罗娟[1] 戢艳琼[1] 路玲俐 张秋芳[1,2] ZHOU Lin;LUO Juan;JI Yan-qiong;LU Ling-li;ZHANG Qiu-fang(Department of Pharmacology,Hubei University of Medicine,Shiyan 442000,China;Hubei Key Laboratory of Wudang Local Chinese Medicine Research,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院药理教研室,湖北十堰442000 [2]湖北医药学院武当特色中药研究湖北省重点实验室,湖北十堰442000

出　　处：《中国病理生理杂志》2021年第4期619-625,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金应急项目(No.81641140,No.81303254);湖北省知识创新专项(自然科学基金)项目(No.2018CFB522);十堰市科学技术研究与开发计划项目(No.2020k36);十堰市引导性科研项目(No.19Y09)。

摘　　要：目的:观察淫羊藿苷(ICA)对异丙肾上腺素(ISO)诱导的H9c2细胞肥大损伤的作用,并进一步探讨其机制是否与内质网应激(ERS)有关。方法:将对数生长期的H9c2细胞随机分为正常对照组、ISO(10μmol/L)组、ISO+ICA(10μmol/L)组、ISO+ICA(20μmol/L)组、ISO+ICA(40μmol/L)组、ISO+4-苯基丁酸(4-PBA,5 mmol/L)组和ISO+美托洛尔(Meto,5μmol/L)组,ICA、4-PBA及Meto预给药1 h,ISO作用48 h。采用MTT法测定细胞活力;检测细胞培养上清液中乳酸脱氢酶(LDH)活性以评价心肌细胞损伤程度;罗丹明标记的鬼笔环肽染H9c2细胞骨架,测定细胞表面积;Hoechst 33258和TUNEL染色测定细胞凋亡率;Western blot检测ERS标志蛋白葡萄糖调节蛋白78(GRP78)、GRP94和CCAAT/增强子结合蛋白同源蛋白(CHOP)的表达。结果:ICA能显著抑制ISO诱导的H9c2细胞肥大及凋亡,减少心肌细胞表面积及LDH的释放,降低细胞凋亡率,下调GRP78、GRP94和CHOP的表达。结论:淫羊藿苷能够有效抑制ISO诱导的H9c2细胞肥大及凋亡,其机制可能与抑制ERS相关信号通路有关。AIM:To investigate the effect of icariin(ICA)on H9c2 cell hypertrophy injury induced by isopro⁃terenol(ISO),and whether its mechanism is related to inhibiting endoplasmic reticulum stress(ERS).METHODS:Cultured H9c2 cells in logarithmic growth phase were randomly divided into control group,ISO group(10μmol/L ISO for 48 h),ISO+ICA groups(10,20 and 40μmol/L ICA+10μmol/L ISO for 48 h),ISO+4-phenylbutyric acid(4-PBA)group(5 mmol/L 4-PBA+10μmol/L ISO for 48 h)and ISO+metoprolol(Meto)group(5μmol/L Meto+10μmol/L ISO for 48 h).Cell viability was measured by MTT assay.Lactate dehydragenase(LDH)activity was detected for evaluating the cell damage.Rhodamine-labeled phalloidin staining was used to measure cell surface area.Apoptosis was analyzed by Hoechst 33258 staining and TUNEL staining.The levels of ERS-related proteins were determined by Western blot.RE⁃SULTS:Compared with control group,the cell viability in ISO group was decreased,while the release of LDH,the cell surface area and the expression of ERS markers glucose-regulated protein78(GRP78),GRP94 and CCAAT/enhancerbinding protein homologous protein(CHOP)were increased(P<0.01).Compared with ISO group,ICA pretreatment in⁃creased cell viability,but decreased the LDH release,the cell surface area,the apoptotic rate,and the expression of GRP78,GRP94 and CHOP.In addition,the effect of ERS inhibitor 4-PBA was similar to ICA.CONCLUSION:Icariin pretreatment attenuates ISO-induced hypertrophy of H9c2 cells,and its mechanism may be related to down-regulating the expression of ERS-related proteins.

关 键 词：淫羊藿苷 异丙肾上腺素 H9C2细胞 内质网应激 心肌肥大 

分 类 号：R541[医药卫生—心血管疾病] R363.2[医药卫生—内科学]