目的 验证通过抑制p75 NTR(p75 neurotrophin receptor)可降低急性高眼压(acute ocular hypertension,AOH)对视网膜神 经节细胞(retinal ganglion cells,RGCs)的损伤作用.方法 应用大鼠急性高眼压模型,玻璃体腔注射p75 NTR抑制剂(TAT-Pep5),按建模后1、3、5d取材,分为正常对照组、假手术组、急性高眼压组、急性高眼压+TAT-Pep5组.采用免疫荧光技术、Westernblotting等方法检测急性高眼压模型中相关蛋白的表达变化.TdT介导的dUTP缺口末端标记(TUNEL)染色检测细胞凋亡情况.结果 急性高眼压模型视网膜Müller细胞上proNGF表达增加.注射p75 NTR抑制剂TAT-Pep5后急性高眼压视网膜上cleaved-caspase 3蛋白量减少,并且RGCs凋亡数目减少.结论 视网膜神经节细胞损伤可引起proNGF表达增加,选择性阻断Müller细胞上的p75 NTR或干预其信号传导通路,可以减少急性高眼压模型中RGCs凋亡.%Objective To explore the role of p75 NTR inhibitor on suppressing retinal ganglion cells (RGCs) apoptosis in a rat model of acute ocular hypertension.Methods The acute ocular hypertension (AOH) rat model was established,and animals were divided into control (Ctrl),sham operation,AOH,AOH + TAT-Pep5 treated groups (1,3 and 5d subgroups).Immunofluorescent staining was performed to detect the expression of proNGF and p75NTR.Western blotting was used to detect the protein expression levels of proNGF and cleaved-caspase 3.TUNEL assay was used to detect cell apoptosis.Results The expression of proNGF was increased in acute ocular hypertension model.When the p75NTR was blocked by its inhibitor (TAT-Pep5),the number of death RGCs (TUNEL positive cell number) were less than that of AOH group.The results of Western blotting showed that the protein expression levels of cleaved-caspase 3 could be up-regulated in retina from AOH group or down-regulated in retina from AOH + TAT-Pep5 group when compared with that of Ctrl and AOH groups.Conclusion RGCs injury can increase proNGF expression.p75NTR blockade can potentiate the survival of RGCs.
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