Calpain抑制剂ALLN对H2O2诱导的661W细胞损害的影响  

Possible effect of N-acetyl-leu-leu-norleucinal,an inhibitor of Calpain,on H_2O_2-induced damage in 661W cells

作　　者：方丽君[1,2] 林颖彬 黄恩 黄天文[2,4] 

机构地区：[1]福建医科大学附属协和医院眼科,福建福州350001 [2]福建省老年医学研究所,福建福州350001 [3]加拿大渥太华大学医学院细胞分子医学系,渥太华K1H8M5 [4]福建医科大学附属协和医院神经内科,福建福州350001

出　　处：《中国药理学通报》2018年第2期181-185,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81100812); 福建省自然科学基金资助项目(No2017J01292)

摘　　要：目的探讨ALLN(N-acetyl-leu-leu-norleucinal)在过氧化氢(H_2O_2)对661W光感受器细胞的毒性损害过程中的可能影响。方法 MTT法测定H_2O_2对661W细胞活性的影响;Western blot法检测在H_2O_2作用661W光感受器细胞中不同时间点Calpain 1和Calpain 2的蛋白水平。结果 MTT结果表明,H_2O_2作用于661W细胞24 h后,661W细胞数明显下降;100μmol·L^(-1)的H_2O_2作用于661W细胞12、18、24 h,Calpain 1和Calpain 2的蛋白水平随H_2O_2作用时间的延长而增加;MTT法测定661W细胞活性,发现ALLN+H_2O_2组细胞活性明显比单纯H_2O_2组高;MTT法检测不同浓度ALLN组(25、50、100、200μmol·L^(-1))的细胞活性,结果显示ALLN组细胞活力与空白对照组无明显差别。结论Calpain抑制剂ALLN可减轻H_2O_2对661W细胞的毒性损害作用,为光感受器细胞氧化损伤及保护提供了实验依据。Aim To investigate the possible effect of N-acetyl-leu-leu-norleucinal( ALLN),an inhibitor of Calpain,on H_2O_2-induced damage in 661 W cells.Methods The cellular survival of 661 W treated with different doses of H_2O_2 without or with ALLN for 24 h was measured by MTT assay. The protein level of Calpain 1 and Calpain 2 was assessed by Western blot.Results Upon the H_2O_2 treatment at the concentrations of 50,100,500,1 000 μmol·L^(-1),the survival rate of 661 W significantly decreased in a dose-dependent manner compared to that of the control group. Furthermore,the protein level of Calpain 1 and Calpain 2 showed an obviously time-dependent increase in 661 W cells treated with 100 μmol·L^(-1) H_2O_2 for 12,18,24 h. Finally, the survival rate of 661 W treated with ALLN and H_2O_2 was higher than that treated only with H_2O_2,and there was no difference in survival rate between ALLN groups( at the concentrations of 25,50,100,200 μmol · L^(-1)) and control group. Conclusions Calpain is involved in the damage induced by H_2O_2. ALLN,the inhibitor of Calpain,attenuates the oxidative damage,which plays a promising protective role in photo-receptor cells under oxidative stress.

关 键 词：ALLN 661W H2O2 Calpain蛋白 光感受器细胞 视网膜色素变性 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]