Chloride channel protein 2 prevents glutamate-induced apoptosis in retinal ganglion cells

Document Type: Original Article

Authors

1 Department of Ophthalmology, The Second Hospital of Jilin University, Jilin University, Changchun, Jilin, 130041, P.R. China

2 Department of Ophthalmology, The China-Japan Union Hospitial of Jilin University, Jilin University, Changchun, Jilin, 130033, Xiantai Street No. 126, Jilin Province, China

3 Department of Colon and Anal Surgery, the First hospital of Jilin University, Jilin University, Changchun, Jilin, 130021, P.R. China

Abstract

Objective(s): The purpose of this study was to investigate the role of chloride channel protein 2 (ClC-2) in glutamate-induced apoptosis in the retinal ganglion cell line (RGC-5).
Materials and Methods: RGC-5 cells were treated with 1 mM glutamate for 24 hr. The expression of ClC-2, Bax, and Bcl-2 was detected by western blot analysis. Cell survival and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, respectively. Caspase-3 and -9 activities were determined by a colorimetric assay. The roles of ClC-2 in glutamate-induced apoptosis were examined by using ClC-2 complementary deoxyribonucleic acid (cDNA) and small inference ribonucleic acid (RNA) transfection technology.
Results: Overexpression of ClC-2 in RGC-5 cells significantly decreased glutamate-induced apoptosis and increased cell viability, whereas silencing of ClC-2 with short hairpin (sh) RNA produced opposite effects. ClC-2 overexpression increased the expression of Bcl-2, decreased the expression of Bax, and decreased caspase-3 and -9 activation in RGC-5 cells treated with glutamate, but silencing of ClC-2 produced opposite effects.
Conclusion: Our data suggest that ClC-2 chloride channels might play a protective role in glutamate-induced apoptosis in retinal ganglion cells via the mitochondria-dependent apoptosis pathway.

Keywords


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