前列地尔对体外培养类缺血再灌注模型视网膜神经节细胞的自噬影响

AUTOPHAGY EFFECT OF ALPROSTADILON CULTURED RETINAL GANGLION CELLS IN A MODEL OF ISCHEMIA-LIKE-REPERFUSION

目的:通过观察前列地尔对体外培养类缺血再灌注模型视网膜神经节细胞(RGC)自噬的影响,探讨其产生保护作用的机制。方法:以原代培养Sprague-Dawley大鼠RGC为研究对象,将培养的细胞随机分为对照组、模型组、给药组。建立类缺血再灌注损伤模型,给药组在建立模型前后加入浓度为45μg/L前列地尔。采用荧光定量PCR方法检测各组细胞中ULK1、Beclin1、Bcl-2mRNA的相对表达量。结果:①与对照组相比,ULK1mRNA、Beclin1mRNA表达均明显升高,其中模型组(P<0.01),给药组(P<0.05);而给药组ULK1mRNA、Beclin1mRNA表达较模型组明显下调(P<0.01)。②模型组与对照组相比,Bcl-2mRNA表达差异无统计学意义(P>0.05);给药组与对照组相比,Bcl-2mRNA表达升高(P<0.05),与模型组相比表达也明显升高(P<0.01)。结论:前列地尔对体外培养类缺血再灌注模型RGC可能通过抑制细胞自噬作用,进而发挥其保护细胞的作用。

Objective： To evaluate the autophagy effect of alprostadil on the cultured retinal ganglion cells （RGC） in a model of ischemia-like-reperfusion in order to investigate the possible protective mechanism of it. Methods： The primary cultured RGC from the Sprague-Dawley rats were randomly divided into groups control, model and alprostadil. The model of ischemia-like-reperfusion was established. Alprostadil was applied at a concentration of 45μg/L before and after establishment of the model in group alprostadil. The relative expression of ULK1, Beclinl and Bcl-2 mRNA in the three groups was evaluated by real-time PCR. Results： As compared with group control, the expression of ULK1 mRNA and Beclinl mRNA was increased in groups model and alprostadil （ P〈0.01 and P 〈0.05 ; respectively）, while the expression of ULK1 mRNA and Beclinl mRNA was less in group alprostadil than in group model （ P 〈0.01）. There was no significant difference in the Bcl-2 mRNA expression between groups control and model （P 〈0.05）, but the expression of Bcl-2 mRNA in group alprostadil was higher than that in groups control （ P 〈0.05） or model （ P 〈0.01）. Conclusion： Alprostadil probably inhibits the autophagy effect on the cul- tured retinal ganglion cells in the model of ischemia-like-reperfusion then plays a role of protection.