钙蛋白酶在TOCP诱导人神经母细胞瘤SH-SY5Y细胞自噬中的调节作用  

The regulatory role of calpain in TOCP-induced autophagy of human neuroblastoma cells SH-SY5Y

作　　者：石晟源 肖双 胡泽慧 龙鼎新 SHI Shengyuan;XIAO Shuang;HU Zehui;LONG Dingxin(School of Public Health,Hengyang Medical School,University of South China,Hunan Province Key Laboratory of Typical Environmental Pollution aud Health Hazards,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学衡阳医学院公共卫生学院典型环境污染与健康危害湖南省重点实验室,湖南衡阳421001

出　　处：《中南医学科学杂志》2023年第3期313-318,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金项目(81673227)。

摘　　要：目的探究钙蛋白酶对三邻甲苯基磷酸酯(TOCP)诱导的人神经母细胞瘤SH-SY5Y细胞自噬的调节作用。方法不同浓度TOCP处理未分化和分化的SH-SY5Y细胞,用荧光分光光度法检测钙蛋白酶活性。抑制钙蛋白酶活性后,通过Western blotting检测TOCP对自噬相关蛋白的影响;抑制钙通道后,用荧光分光光度法检测胞质钙离子浓度对钙蛋白酶活性的影响。结果TOCP诱导未分化和分化的SH-SY5Y细胞钙蛋白酶活性上升;钙蛋白酶抑制剂可降低TOCP诱导的自噬相关蛋白LC3-II和Beclin1表达。维拉帕米和2-APB阻断钙通道后钙蛋白酶活性降低,维拉帕米而非2-APB可以抑制TOCP诱导的自噬相关蛋白LC3-II表达。结论TOCP在未分化和分化的SH-SY5Y细胞中诱导的自噬受钙蛋白酶活性的调节。Aim To investigate the regulatory role of calpain on TOCP-induced autophagy.Methods Undifferentiated and differentiated SH-SY5Y cells were used as experimental objects,and the calpain activity was detected by fluorescence spectrophotometry after treatment with gradient concentrations of TOCP.The effect of TOCP on autophagy-related proteins were detected by Western blotting after the inhibition of calpain activity.Effect of cytoplasmic calcium ions on calpainase activity by fluorescence spectrophotometry after inhibition of calcium channels.Results TOCP increased calpain activity in both undifferentiated and differentiated SH-SY5Y cells.Calpain inhibitor reduced the expression of autophagy-related proteins LC3-II and Beclin1 induced by TOCP.Blocking calcium channels with verapamil and 2-APB reduced calpain activity.Verapamil but not 2-APB inhibited the expression of LC3-II induced by TOCP.Conclusion TOCP-induced autophagy in undifferentiated and differentiated SH-SY5Y cells is regulated by calpain activity.

关 键 词：自噬 钙蛋白酶 SH-SY5Y细胞系 TOCP 

分 类 号：R114[医药卫生—卫生毒理学] X592[医药卫生—公共卫生与预防医学]