过氧化物还原酶I-硫氧还蛋白、超氧化物歧化酶及过氧化氢酶在大鼠肝脏缺血再灌注损伤中的表达变化

Expression of peroxiredoxin I-thioredoxin,superoxide dismutases and catalases mRNA in hepatic ischemia-reperfusion injury of rats

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 expression change of peroxiredoxin I-thioredoxin （PrxI-Trx）, superoxide dismutases （SOD） and catalases （CAT） in hepatic Ischemia-Reperfusion injury of rats, and to explore their role in oxidative-stress response. Methods The hepatic ischemia-reperfusion injury model of rats was established by clipping the hepatic blood vessel released both hepatic left and middle lobes as well as bile duct pedicle with non-damage vascular clamp for 30 minutes. The serum and hepatic tissues were taken 6 hours after clipping. The morphological changes were observed under a microscope using HE staining. The serum alanine aminotransferase（ALT） was detected by Automatic biochemical analyzer. The PrxI-Trx, SOD and CAT mRNA expression were evaluated by RT-PCR. The protein level of PrxI, SOD and CAT were estimated by Western blotting. Results Compared with the control group, the ALT level and HE results of hepatic ischemia-reperfusion injury group showed that liver cells were significantly impaired. The PrxI-Trx, CAT and SOD mRNA expression were increased significantly. The protein level of PrxI, SOD and CAT also increased significantly. Conclusion PrxI-Trx, SOD and CAT may play an important role in inhibiting the oxidative-stress response induced by ischemia-reperfusion injury.