The effects of erythropoietin on RhoA/Rho-associated kinase expression in rat retinal explants cultured with glutamate.
The effects of erythropoietin on RhoA/Rho-associated kinase expression in rat retinal explants cultured with glutamate.
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DOI：
10.3892/mmr.2012.947
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发表时间：
2012-09
影响因子：
3.4
通讯作者：
Ping Huang;Jing Wang;Xi Shen;Qin Jiao;Y. Cheng;B. Xie;Y. Zhong
中科院分区：
文献类型：
--
作者：
Ping Huang;Jing Wang;Xi Shen;Qin Jiao;Y. Cheng;B. Xie;Y. Zhong
本研究旨在探讨促红细胞生成素（EPO）对谷氨酸诱导的大鼠视网膜RhoA/Rho相关激酶（ROCK）表达的影响。将视网膜外植体在无血清R16营养培养基中培养24 h后，将视网膜外植体分为对照组（R16营养培养基）、谷氨酸组（R16营养培养基含有5mM/l谷氨酸）和谷氨酸+EPO组（R16营养培养基含有5mM/l谷氨酸和6.0U/ml EPO），并继续培养72 h。RT-PCR和Western blotting检测视网膜组织中总RhoA、ROCK 1和ROCK 2的mRNA和蛋白表达，GST-RBD结合和免疫印迹检测视网膜组织中活性RhoA。总RhoA mRNA和蛋白表达在对照组、谷氨酸组和谷氨酸+EPO组之间没有显著差异。谷氨酸可增加视网膜组织中活性RhoA、ROCK 1和ROCK 2的表达（P <0.05），而谷氨酸+EPO组活性RhoA、ROCK 1和ROCK 2的表达明显低于单纯谷氨酸组（P <0.05），与对照组相似。总之，EPO下调活性RhoA，ROCK1和ROCK2的表达与谷氨酸培养的视网膜外植体。
The aim of this study was to investigate the effects of erythropoietin (EPO) on RhoA/Rho-associated kinase (ROCK) expression in rat retinal explants cultured with glutamate. After the retinal explants were cultured in serum-free R16 nutrient medium for 24 h, the retinal explants were divided into control (R16 nutrient medium), glutamate (R16 nutrient medium containing 5 mM/l glutamate) and glutamate+EPO (R16 nutrient medium containing 5 mM/l glutamate and 6.0 U/ml EPO) groups, and culturing was continued for another 72 h. The mRNA and protein expression of total RhoA, ROCK1 and ROCK2 in the retinal explants was examined by RT-PCR and western blotting, and active RhoA in the retinal explants was detected via GST-RBD binding and immunoblotting with an antibody specific to active RhoA. The total RhoA mRNA and protein expression did not differ substantially between the control, the glutamate and the glutamate+EPO groups. The glutamate increased the active RhoA, ROCK1 and ROCK2 expression in cultured retinal explants (P<0.05), whereas the expression of active RhoA, ROCK1 and ROCK2 in the glutamate + EPO group was significantly lower than that in the simple glutamate group (P<0.05) and similar to that in the control group. In conclusion, EPO downregulates active RhoA, ROCK1 and ROCK2 expression in retinal explants cultured with glutamate.