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目的观察过氧化物还原酶I-硫氧还蛋白(PrxI-Trx)、超氧化物歧化酶(SOD)及过氧化氢酶(CAT)抗氧化体系在大鼠肝脏缺血再灌注损伤模型中的表达变化,探讨其在抗氧化应激反应中的作用。方法通过无损伤血管夹钳夹通往大鼠肝左叶、肝中叶的血管和胆管蒂,30 min后松开血管夹,制造大鼠70%肝脏缺血再灌注损伤模型。损伤再灌注6h后取血和肝脏。全自动生化分析仪测丙氨酸氨基转移酶(ALT)含量。HE法观察大鼠肝脏形态学改变。采用RT-PCR的方法观察在肝脏缺血再灌注损伤中PrxI-Trx、SOD、CAT氧化还原体系mRNA水平的表达变化。采用Western blotting测定PrxI、SOD和CAT的蛋白表达水平。结果与对照组相比,血清中ALT水平和HE结果均显示肝脏缺血再灌注损伤组大鼠肝细胞明显受损。PrxI-Trx、SOD和CAT的mRNA水平明显升高。同时,PrxI、SOD和CAT的蛋白表达水平也明显升高。结论 PrxI-Trx、SOD和CAT在肝脏缺血再灌注损伤中均发挥了抗氧化应激作用,对肝细胞具有保护作用。
Objective To observe the effects of peroxiredoxin I-thioredoxin (PrxI-Trx), superoxide dismutase (SOD) and catalase (CAT) antioxidant system on hepatic ischemia-reperfusion injury in rats Expression changes, to explore its role in the anti-oxidative stress response. Methods The blood vessels and bile duct pedicels leading to the left lobe of liver and middle lobe of liver were ligated without injury and the vessel clips were released 30 min later to establish a model of 70% hepatic ischemia-reperfusion injury in rats. Blood and liver were harvested 6h after injury and reperfusion. Automatic biochemical analyzer measured alanine aminotransferase (ALT) content. HE staining was used to observe the changes of rat liver morphology. The changes of mRNA expression of PrxI-Trx, SOD and CAT redox system in liver ischemia-reperfusion injury were observed by RT-PCR. The protein expression levels of PrxI, SOD and CAT were determined by Western blotting. Results Compared with the control group, the level of ALT in serum and the results of HE all showed that the hepatocytes of rats with liver ischemia-reperfusion injury were obviously damaged. PrxI-Trx, SOD and CAT mRNA levels were significantly increased. At the same time, PrxI, SOD and CAT protein expression levels also significantly increased. Conclusion PrxI-Trx, SOD and CAT exert anti-oxidative stress in liver ischemia-reperfusion injury and have a protective effect on hepatocytes.