大鼠肾缺血再灌注损伤模型心肌内过氧化酶Ⅲ的表达变化

Expression changes of peroxiredoxin Ⅲ in the myocardium of renal ischemia reperfusion injury model in rats

目的观察过氧化酶Ⅲ(peroxiredoxinⅢ,PrxⅢ)在大鼠肾缺血再灌注损伤(renal ischemia reperfusion injury,RIRI)模型心肌内的表达变化,探讨PrxⅢ在抗氧化应激反应中的作用。方法大鼠切除右肾后,无损伤动脉夹夹闭左肾动脉,建立肾缺血再灌注损伤模型。再灌注24h后取材(血、肾、心)。采用酶偶联速率法和苦味酸法检测血尿素氮(blood urea nitrogen,BUN)、肌酐(seaum creatinine,SCr)浓度;HE染色法观察大鼠肾组织形态;硫代巴比妥酸比色法(thiobarbituric acid,TBA)检测心肌丙二醛(Malondialdehyde,MDA)含量;应用逆转录聚合酶链反应法和免疫印迹法测定大鼠心肌组织内抗氧化酶PrxⅢ的mRNA与蛋白表达水平。结果 RIRI组大鼠血清中BUN和SCr浓度明显高于对照组,RIRI组大鼠心肌组织中MDA含量、PrxⅢmRNA水平相对表达量、PrxⅢ蛋白水平相对表达量明显高于对照组,差异有统计学意义(P<0.05)。结论 RIRI后,心肌组织出现过氧化损伤;PrxⅢ在肾缺血再灌注损伤大鼠的心组织内发挥了抗氧化应激的作用。

Objective To observe the expression of peroxiredoxin Ⅲ （Prx Ⅲ） in renal ischemia reperfUsion injury model rats and study the role in oxidative stress response. Methods After removal of the right kidney, the left renal artery without injury was removed, and the model of renal ischemia and reperfusion injury was established. After 24 hours of reperfusion, the blood and kidney were taken. The serum blood urea nitrogen（BUN） and serum creatinine（SCr） were detected with pierie acid method and enzyme coupling rate method, the malondialdehyde （MDA） content in myocardial tissue was determined by thiobarbituric acid eolorimetric method. The renal morphology change was observed by HE staining. The Prx Ⅲ mRNA and protein expression in myocardium were evaluated by reverse transcription polymerase chain reaction（RT- PCR） and western blot. Results Compared with the control group, the BUN and SCr in serum of renal ischemia reperfusion injury group were significantly increased. The MDA content, the PrxⅢ mRNA relative expression level and protein relative expression level in myocardium of renal ischemia reperfusion injury group were higher than that of control group,there were statistically significant difference （P〈 0. 05）. Conclusion After renal ischemia reperfusion injury, myocardial tissue appeared oxidative damage. PrxⅢ plays a role of anti oxidative stress in the heart tissue of rats with renal ischemia reperfusion injury.