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目的探讨内皮素-1(ET-1)、ET-2、ET-3、内皮素受体A(ETRA)、ETRB和内皮素转换酶(ECE)在8周糖尿病SD大鼠视网膜中的表达。方法采用限制性片段差异显示PCR(RFDD-PCR)技术建立正常和8周糖尿病大鼠视网膜基因差异表达谱,并以半定量RT-PCR和蛋白免疫印迹方法分别对两组间表达存在差异的6个基因(ET-1、ET-2、ET-3、ETRA、ETRB、ECE)的表达进行验证。结果 RFDD-PCR结果显示,糖尿病组ET-1、ET-2、ET-3、ETRA、ETRB和ECE表达上调。半定量RT-PCR结果和蛋白免疫印迹结果显示,6种基因和蛋白在糖尿病组的表达(相对光密度比值)高于正常组,表达差异具有统计学意义(P均<0.05,P均<0.01)。结论 ET-1、ET-2、ET-3、ETRA、ETRB和ECE在糖尿病视网膜中呈高表达,提示这6种基因可能参与了糖尿病视网膜的发病过程。
Objective To investigate the expression of endothelin-1, ET-2, ET-3, endothelin receptor A (ETRA), ETRB and endothelin-converting enzyme (ECE) in the retina of 8-week diabetic SD rats. Methods The differential expression profiles of retinal genes in normal and 8-week diabetic rats were established by RFDD-PCR. Semi-quantitative RT-PCR and Western blotting were used to detect the expression differences of retinal genes between the two groups (ET-1, ET-2, ET-3, ETRA, ETRB, ECE). Results The results of RFDD-PCR showed that the expressions of ET-1, ET-2, ET-3, ETRA, ETRB and ECE in diabetic group were up-regulated. Semi-quantitative RT-PCR results and Western blotting results showed that the expression of six genes and proteins in diabetic group (relative optical density ratio) was higher than that in normal group (P <0.05, P <0.01 respectively) ). Conclusions ET-1, ET-2, ET-3, ETRA, ETRB and ECE are highly expressed in the diabetic retina, suggesting that these six genes may be involved in the pathogenesis of diabetic retinopathy.