Objective  The aim of this study was to investigate the RhoA/ROCK expression effects of EPO on rat retinal explants cultured with glutamate.
Methods  After the retinal explants were cultured in serum-free R16 nutrient medium for 24 hours, the retinal explants were divided into the control group (R16 nutrient medium), the glutamate group (R16 nutrient medium containing 5 mM/L glutamate) and the glutamate + EPO group (R16 nutrient medium containing5 mM/L glutamate and 6.0 U/ml EPO), and continue cultured for another 72 hours. The mRNA and protein expression of total RhoA, ROCK1 and ROCK2 inretinal explants was examined by RT-PCR and Western blotting, the active RhoA in retinal explants was detected via GST-RBD binding and immunoblotting with antibody specific to active RhoA.
Results  The total RhoA mRNA and protein expression did not differ substantially among the control group, the glutamate group and the glutamate + EPO group. The glutamate could increase the active-RhoA, ROCK1, ROCK2 expression in cultured retinal explants (P<0.05), and the expression of active-RhoA, ROCK1, ROCK2 inthe glutamate +EPO group was significant lower than that in the simple glutamate group (P<0.05), and closed to the control group.
Conclusion  EPO could down-regulate the active-RhoA, ROCK1, ROCK2 expression in retinal explants cultured with glutamate