摘要
目的分析氧化应激损伤的HEI-OC1毛细胞中热休克蛋白75/78(GRP75/78)的表达水平,探讨GRP75、GRP78在耳蜗毛细胞凋亡过程中的可能作用。方法用不同浓度(20、40、60μM)叔丁基过氧化氢(tBHP)染毒构建HEI-OC1毛细胞株凋亡模型,对照组不加t-BHP,培养24h后用Western blot检测各组毛细胞凋亡标志物Caspase-3、Active-Caspase-3表达水平,实时荧光定量PCR检测GRP75、GRP78mRNA表达水平。结果与对照组相比,各染毒组Caspase-3和Active-Caspase-3蛋白表达水平显著增高(F=1 196.741,P<0.001和F=184.131,P<0.001),40μM和60μM染毒组Caspase-3和Active-Caspase-3蛋白表达水平显著高于20μM染毒组(P<0.01),但40μM和60μM染毒组之间差异无统计学意义(P>0.05)。与对照组相比较,各染毒组GRP75和GRP78mRNA表达水平显著增高(F=228.065,P<0.001和F=1 298.057,P<0.001),40μM和60μM染毒组GRP75和GRP78mRNA表达水平显著高于20μM染毒组(P<0.01),但40μM和60μM染毒组之间差异无统计学意义(P>0.05)。结论 GRP75和GRP78在氧化应激损伤的耳蜗毛细胞中有抗凋亡的生物学作用。
Objective To study the biological functions of GRP75/78 in the apoptosis of cochlear hair cells by analyzing the expression level of GRP75/78 in apoptotic HEI-OC1 cells induced by oxidative stress.Methods Using different concentration(20μM,40μM,60μM)of tert-Butyl hydroperoxide(t-BHP)canister to cultivate HEI-OC1 cells to constructe apoptotic cell model.The control group didn't add t-BHP.Western blot was used to analyze the expression level of Caspase-3 and active-Caspase-3 as markers of apoptosis.Quantitative Real-time PCR was used to analyze the expression level ofGRP75 and GRP78 mRNA.Results The expression level of Caspase-3 and active-Caspase-3 of all exposure groups significantly increased(F=1 196.741,P<0.01 and F=184.131,P<0.01).The expression level of Caspase-3 and active-Caspase-3 of 40μM and 60μM exposure groups was significantly higher than that of 20μM exposure group(P<0.01).But between the 40μM and 60μM exposure groups,the expression level of Caspase-3 and active-Caspase-3 showed no significant differences(P>0.05).The expression level ofGRP75 and GRP78 mRNA of all exposure groups significantly increased(F=228.065,P<0.01和F=1 298.057,P<0.01).The expression level ofGRP75 and GRP78 mRNAof 40μM and 60μM exposure groups was significantly higher than that of 20μM exposure group(P<0.01).But between the 40μM and 60μM exposure groups,the expression level ofGRP75 and GRP78 mRNAhad no significant differences(P>0.05).Conclusion GRP75 and GRP78 play an anti apoptotic biological role in cochlear hair cells damaged by oxidative stress.
作者
张菲玲
夏云
姚裕恒
王军义
Zhang Feiling;Xia Yun;Yao Yuheng;Wang Junyi(Guangdong Pharmaceut Univ,Sch Public Health,Guangzhou,510310,China)
出处
《听力学及言语疾病杂志》
CAS
CSCD
北大核心
2018年第4期419-422,共4页
Journal of Audiology and Speech Pathology
基金
广东省科技计划项目(2014A0202126)
广东省2017年大学生创新创业训练计划项目(201710573034)
