Abstract
Purpose :
Glaucoma is the second leading cause of blindness. Apoptosis of retinal ganglion cells (RGCs) is an accepted important cause of glaucomatous optic injury.Rho kinase expression significantly increased in the RGCs apoptosis.This study aimed to explored the effects of RhoA ,a Rho GTPase,on the apoptosis/survival of RGCs and the involved pathway, further to suggest a potential application for therapy.
Methods :
The RGCs in vitro were treated with siRhoA for 24 hours to silence RhoA. Quantitative RT-PCR was performed to evaluate the effects of siRNA on RhoA expression. 200μM of H2O2 was used for 1h to induce oxidative stress. Apoptosis of RGC cells under oxidative stress was quantified in-situ using the TUNEL assay and flow cytometry.The mRNA expressions of RhoA, Nogo receptor , Caspase3 anf Bcl-2 were evaluated by quantitative RT-PCR. Western Blot was used to measure the protein levels of RhoA,ROCK1, ROCK2, Nogo Receptor, Casepase-3 and Bcl-2.
Results :
siRhoA can efficiently inhibit RhoA expression in RGCs. Furthermore, RhoA siRNA Treatment Protects against H2O2 -induced Apoptosis. By slience RhoA,we showed that ROCK1, ROCK2, Nogo Receptor, Casepase-3 expression levels were significantly decreased while the expression of Bcl-2 was siginificantly increased.
Conclusions :
siRhoA reduced the ROCK1, ROCK2, Nogo Receptor, Casepase-3 expression, and increased the Bcl-2 expression, to prevent H2O2-induced apoptosis in RGC cells. The results suggest that siRhoA is an effective neuroprotection for RGCs through reduce apoptosis.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.