CYLD干扰对实验性视网膜脱离中光感受器细胞坏死性凋亡伴发自噬的影响

Effects of cylindromatosis interference on autophagy in necroptosis of photoreceptor cells in experimental retinal detachment

目的探讨圆柱瘤基因(CYLD)干扰对实验性视网膜脱离中光感受器细胞坏死性凋亡伴发自噬的影响。方法大鼠视网膜下注射慢病毒载体3周后,在各实验组建立视网膜脱离的模型,分为正常对照组、未干预组、CYLD干预组、阴性干预组。建模3 d后,使用电镜观察光感受器细胞的形态学特点并判断其死亡方式。采用Western blot法检测视网膜裂解的LC3蛋白、beclin-1蛋白表达。结果电镜检测结果显示CYLD干预组光感受器细胞坏死数明显低于未干预组和阴性干预组(P<0.05)。Western blot结果表明CYLD干预组LC3蛋白、beclin-1蛋白表达水平显著低于未干预组和阴性干预组(P<0.01)。结论电镜检测和Western blot检测结果共同显示CYLD干扰可以通过抑制LC3和beclin-1的活化,从而抑制光感受器细胞的坏死性凋亡伴发的自噬。

Objective To evaluate the effects of cylindromatosis(CYLD)on the autophagy in necroptosis of photoreceptor cells in experimental retinal detachment.Methods Three weeks after subretinal injection of lentivirus CYLD shRNA,retinal detachment model was established in each group.The rats were divided into the following four groups:attached group,untreated group,sh-CYLD group,and sh-neg group.Three days after modeling,the morphological characteristics and death pattern of photoreceptor cell were observed by electron microscopy.Cleaved protein LC3 and Beclin-1 were measured by Western blot.Results Three days after modeling,transmission electron microscopy showed that necrosis of photoreceptor cells significantly decreased in the sh-CYLD group compared with that in the untreated group and the sh-neg group(P<0.05).Western blot showed that the protein expression of cleaved protein LC3 and Beclin-1 significantly decreased in the sh-CYLD group compared with that in the untreated group and the sh-neg group(P<0.01).Conclusion The results of transmission electron microscopy and Western blot show that CYLD interference can protect photoreceptors from autophagy in necroptosis through inhibit the activation of LC3 and Beclin-1.