前列地尔对原代培养神经元类缺血再灌注损伤后线粒体解耦联蛋白4表达的影响

Effects of Alprostadil on UCP4 expression in primary cultured rat′s neurons induced by ischemia-like-reperfusion

目的:探讨线粒体解耦联蛋白4(UCP4)在原代培养神经元类缺血再灌注损伤后不同时间点的表达变化及前列地尔对其影响。方法:以原代培养神经元为研究对象,建立类缺血再灌注损伤模型,应用免疫细胞化学方法观察类缺血再灌注后3 h,6 h,12 h时UCP4的表达变化。实验组为在缺血期及再灌期均给予0.01 mg/mL前列地尔,观察前列地尔对UCP4表达的影响。结果:UCP4阳性细胞随类缺血再灌注时间的延长,阳性神经元胞浆的灰阶值数值变大,染色变浅,反映神经细胞内UCP4蛋白表达量减少。在再灌注3 h时UCP4的表达即降低;再灌注6 h时UCP4的表达明显降低,再灌注12 h时其表达仍在下降。在再灌注后3 h,6 h,12 h组与正常对照组比较,差异具有显著性意义(P<0.05),实验组UCP4阳性细胞胞浆的灰阶值在再灌注后3 h,6 h,12 h时均低于类缺血再灌注模型组,差异有显著性(P<0.05)。结论:UCP4在类缺血再灌注后不同时间点的表达呈动态变化,提示UCP4可能参与了脑缺血再灌注损伤的病理过程;而前列地尔可上调UCP4的表达,从而发挥保护神经细胞的作用。

Objective：To study the expression of UCP4 in primary cultured rat＇s cortical neurons induced by ischemia-like-reperfusion and influence of the Alprostadil on it. Methods ：Ischemia-like-reperfusion was made on primary cultured rat＇s neurons. Immunoh istochemical method was used to investigate the expression of UCP4 after reper- fusion at 3 h,6 h,12 h. Alprostadil group was given 0. 01 mg/mL Alpprostadil during ischemia and reperfusion. The expression of UCP4 was investigated in Alprostadil group. Results：With the prolongation of the time in ischemic group and treatment group,intracellular gray-scales of positive neurons increased, the colour lighted,this indicated that the expression of UCP4 decreased. The expression of UCP4 was reduced 3 h after ischemia-like-reperfusion and was reduced significantly 6 h after reperfusion,it still decreased 12 h after ischemia like-reperfusion. Compared with the normal control group,UCP4 was significantly decreased at different time in the model group（P〈0.05）. Intracellular gray-scales of Alprastadjl group was lower than that of the model group at different time（P〈0.05）. Conclusion： The expression of UCP4 was different at different time after ischemia-like-reperfusion. This suggested that UCP4 probably took part in physiological process of ischemia-like-reperfusion. Alprostadil can increase the expression of UCP4 and protect neurons.