摘要
The purpose of this study was to investigate the effect of inhibition of calpain on retinal ganglion cell-5（RGC-5） necroptosis following oxygen glucose deprivation（OGD）. RGC-5 cells were cultured in Dulbecco＇s-modified essential medium and necroptosis was induced by 8-h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. The calpain expression was detected by Western blotting and immunofluorescence staining. The calpain activity was tested by activity detection kit. Flow cytometry was used to detect the effect of calpain on RGC-5 necroptosis following OGD with or without N-acetyl-leucyl-leucyl-norleucinal（ALLN） pre-treatment. Western blot was used to detect the protein level of truncated apoptosis inducing factor（t AIF） in RGC-5 cells following OGD. The results showed that there was an up-regulation of the calpain expression and activity following OGD. Upon adding ALLN, the calpain activity was inhibited and t AIF was reduced following OGD along with the decreased number of RGC-5 necroptosis. In conclusion, calpain was involved in OGD-induced RGC-5 necroptosis with the increased expression of its downstream molecule t AIF.
The purpose of this study was to investigate the effect of inhibition of calpain on retinal ganglion cell-5（RGC-5） necroptosis following oxygen glucose deprivation（OGD）. RGC-5 cells were cultured in Dulbecco＇s-modified essential medium and necroptosis was induced by 8-h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. The calpain expression was detected by Western blotting and immunofluorescence staining. The calpain activity was tested by activity detection kit. Flow cytometry was used to detect the effect of calpain on RGC-5 necroptosis following OGD with or without N-acetyl-leucyl-leucyl-norleucinal（ALLN） pre-treatment. Western blot was used to detect the protein level of truncated apoptosis inducing factor（t AIF） in RGC-5 cells following OGD. The results showed that there was an up-regulation of the calpain expression and activity following OGD. Upon adding ALLN, the calpain activity was inhibited and t AIF was reduced following OGD along with the decreased number of RGC-5 necroptosis. In conclusion, calpain was involved in OGD-induced RGC-5 necroptosis with the increased expression of its downstream molecule t AIF.
基金
supported by grants from National Natural Science Foundation of China(No.81371011,and No.81571939)
Wu Jie-Ping Medical Foundation of the Minister of Health of China(No.320.6750.14118)
Natural Science Foundation of Hunan Province(No.2015JJ2187)
Teacher Research Foundation of Central South University(No.2014JSJJ026)
Science Foundation for the Youth Scholars of Central South University(No.120973)
Project of Innovation-driven Plan of Central South University(No.2015CXS022)
作者简介
Shuang CHEN, E-mail： 405237696@qq.com;Jie Yan, E-mail： 69574720@qq.comCorresponding author, E-mail： xiongkun2001 @ 163.com
