Rho激酶抑制剂Y27632对急性视网膜缺血-再灌注损伤大鼠凋亡相关基因表达的影响

Effect of Rho kinase inhibitor Y27632 on the expressions of apoptosis-related genes in rats with acute retinal ischemia-reperfusion injury

目的探讨Rho激酶抑制剂Y27632对急性视网膜缺血-再灌注损伤大鼠凋亡相关基因表达的影响。方法80只SD大鼠随机分为急性缺血-再灌注组(再灌注组)、生理盐水对照组(生理盐水组)、Y27632治疗组(Y27632组)和正常对照组(正常组),每组20只。生理盐水组和Y27632组右眼玻璃体腔注射生理盐水和稀释后的Y276324μL,随后再灌注组、生理盐水组和Y27632组大鼠右眼构建急性视网膜缺血-再灌注模型,正常组不做任何处理。再灌注后24 h和7 d采用HE染色观察大鼠视网膜结构,再灌注后24 h采用Western blot法检测Caspase-3蛋白表达,实时荧光定量PCR检测Bcl-2 mRNA表达。结果再灌注后24 h,再灌注组和生理盐水组视网膜厚度较Y27632组和正常组增加(P<0.05),Y27632组视网膜厚度较正常组增加(P<0.05);再灌注后7 d,再灌注组和生理盐水组视网膜厚度较Y27632组和正常组减少(P<0.05),Y27632组视网膜厚度较正常组减少(P<0.05)。再灌注组和生理盐水组Caspase-3蛋白表达高于正常组和Y27632组(0.973±0.153和0.969±0.144 vs.0.337±0.053和0.509±0.071)(P<0.05),Y27632组Caspase-3蛋白表达高于正常组(P<0.05)。Y27632组Bcl-2 mRNA表达高于再灌注组、生理盐水组和正常组(2.80±0.37 vs.2.27±0.62、2.28±0.54和1.07±0.23)(P<0.05),再灌注组和生理盐水组Bcl-2 mRNA表达高于正常组(P<0.05)。结论Y27632玻璃体腔注射可以减少急性视网膜缺血-再灌注损伤大鼠视网膜中Caspase-3蛋白表达,增加Bcl-2 mRNA表达。

Objective To investigate the effect of Rho kinase inhibitor Y27632 on the expressions of apoptosis-related genes in the rats with acute retinal ischemia-reperfusion injury.Methods A total of 80 SD rats were randomly and equally divided into four groups.The right eyes of group B and group C were injected with normal saline and diluted Y276324μL into vitreous.Acute retinal ischemia-reperfusion model was established in the right eyes of the rats in group A,group B and group C.Group D was not taken any treatment.At 24 hours and 7 days after reperfusion,the retinal structure of the rats was observed by HE staining.The protein expression of Caspase-3 was detected by Western blot,and mRNA expression of Bcl-2 was detected by real-time quantitative PCR at 24 hours after reperfusion.Results The retinal thickness in groups of A and B was thicker than that in groups of C and D at 24 hours after reperfusion,which was thicker in group C than that in group D(P<0.05).The retinal thickness in groups of A and B was less than that in groups of D and C at 7 days after reperfusion,which was less in group C than that in group D(P<0.05).The protein expression of Caspase-3 in groups of A and B was higher than that in groups of D and C(0.973±0.153 and 0.969±0.144 vs.0.337±0.053 and 0.509±0.071)(P<0.05),which was higher in group C than that in group D(P<0.05).The Bcl-2 mRNA expression was higher in group C than that in groups of A,B and D(2.80±0.37 vs.2.27±0.62,2.28±0.54 and 1.07±0.23)(P<0.05),which was higher in groups of A and B than that in group D(P<0.05).Conclusion Intravitreal injection of Y27632 can reduce the protein expression of Caspase-3 and enhance the mRNA expression of Bcl-2 in the retina of the rats with acute retinal ischemia-reperfusion injury.