目的探讨缺血再灌注(IR)期间小鼠肾组织乙醛脱氢酶2(ALDH-2)表达的变化。方法采用雄性C57BL/6J小鼠夹闭双侧肾蒂的方法建立小鼠肾脏IR损伤模型,肾蒂夹闭时间为30min。采用随机数字表法将小鼠分为7组:假手术组、缺血再灌注1h、3h、6h、12h、24h及72h组,每组8只小鼠。分别于假手术后1d及再灌注后1、3、6、12、24及72h获取各组小鼠肾脏,采用实时荧光定量逆转录聚合酶链反应(Real-timePCR)法检测肾组织ALDH-2mRNA的表达,采用蛋白质印迹法检测肾组织ALDH=2蛋白的表达。结果小鼠肾脏IR后肾组织ALDH-2mRNA的水平迅速升高,至再灌注后3h时达到峰值,与假手术组比较,IR3h组和IR6h组均显著升高(P〈0.001);小鼠肾脏氓后肾组织ALDH-2蛋白表达逐渐降低,至再灌注后3h降至最低,之后逐渐升高,至再灌注后24h达到峰值,与假手术组比较,各瓜组肾组织ALDH-2蛋白表达的差异均有统计学意义(P〈0.01)。结论肾脏IR后ALDH-2在转录和蛋白水平的表达均有显著改变;IR过程对肾脏AL—DH-2的表达产生影响,并间接影响其对肾脏氓后致伤效应的调控作用。
Objective To observe the expression change of aldehyde dehydrogenase 2 (ALDH- 2) in renal tissues of mice under ischemia-reperfusion (IR) injury. Method Male C57BL/6J mice IR injury model was establish by clipping bilateral renal pedicles for 30 min. Fifty-six mice were divided into 7 groups by using the random number table method: control group, and IR 1 h, 3 h, 6 h, 12 h, 24 h and 72 h groups. Kidneys in IR 1 h, 3 h, 6 h, 12 h, 24 h and 72 h groups were harvested at each time point, and those in the control group were harvested at 24 h of sham operation. Total mRNA expression levels of ALDH-2 in the kidneys were detected by real-time PCR. Western blotting was used to detect the ALDH-2 protein expression. Result The mRNA expression levels of ALDH-2 were increased right after IR and reached the peak at 3 h. The expression levels of ALDH-2 mRNA in IR 3 h group and IR 6 h group were significantly higher than in control group (P〈0. 001). The expression of ALDH-2 protein began to decline after IR and reached a minimum at 3 h, then ALDH-2 protein expression was increased and reached its peak at 24 h (P〈0. 01). The ALDH-2 expression in IR groups was significantly lower than in control group ( P 〈 0. 01 or P 〈 0. 001 ). Conclusion The expression of ALDH-2 has a remarkable change after kidney IR, suggesting that the process of IR has an impact on ALDH-2 expression, and indirectly affects the function of regulation of ALDH-2 in kidney IR injury.