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目的:探讨大鼠缺血再灌注后溶酶体组织蛋白酶D(Cathepsin D CD)、caspase-9在不同时段蛋白质及mRNA表达变化。方法:将60只S D大鼠随机分为三组:正常组(10只),假手术组(10只),脑缺血再灌注组(40只),线栓法制备大脑中动脉梗死模型(MCAO),免疫组化及RT-PCR法分别检测Cathepsin D、caspase-9的蛋白和mRNA表达。结果:与正常组和假手术组比较,模型组大鼠脑缺血再灌注损伤后6h Cathepsin D的蛋白和mRNA表达明显增强(P<0.05),24h达高峰,48h仍保存高水平。caspase-9蛋白和mRNA6h开始明显升高,12h达高峰,此后缓慢下降,但48h组仍显著高于对照组(P<0.05),结论:Cathepsin D、caspase-9在大鼠脑缺血再灌注后表达增强,溶酶体可能参与了脑缺血再灌注损伤后神经细胞凋亡的过程。
Objective: To investigate the changes of protein and mRNA expression of cathepsin D (CD) and caspase-9 at different time points after ischemia-reperfusion in rats. Methods: Sixty SD rats were randomly divided into three groups: normal group (10 rats), sham operation group (10 rats), cerebral ischemia reperfusion group (40 rats), middle cerebral artery occlusion model MCAO), immunohistochemistry and RT-PCR were used to detect the expression of Cathepsin D, caspase-9 protein and mRNA. Results: Compared with the normal group and the sham operation group, the expression of Cathepsin D protein and mRNA increased significantly 6 h after cerebral ischemia-reperfusion injury in model group (P <0.05), peaked at 24 h and remained high at 48 h. The protein and mRNA expression of caspase-9 increased significantly at 6h and peaked at 12h, then decreased slowly but remained significantly higher at 48h (P <0.05). Conclusion: Cathepsin D and caspase-9 play roles in cerebral ischemia-reperfusion After the expression increased, lysosomes may be involved in the process of neuronal apoptosis after cerebral ischemia-reperfusion injury.