血红素氧合酶1基因转移对大鼠肾缺血再灌注损伤的防护作用

Heme oxygenase-1 gene transfer protects rat kidney transplant from ischemia/reperfusion injury

目的探讨血红素氧合酶1(HO-1)基因转移对大鼠自体移植肾缺血再灌注损伤的保护作用。方法构建HO-1腺病毒表达载体,经肾动脉灌注转染26只大鼠(实验组)移植肾,4℃保存24h后行自体移植,移植后5d切除对侧肾脏;以25只大鼠为对照。于移植后3h、3d,应用逆转录聚合酶链反应(RT-PCR)及免疫组织化学方法检测移植肾HO-1基因及蛋白的表达;应用酶联免疫吸附试验(ELISA)法测定肾组织匀浆中HO-1蛋白的含量(以吸光度值表示)。结果移植后3h及3d,实验组移植肾HO-1mRNA的表达强度分别为0·65±0·11及0·86±0·17,而对照组分别为0·09±0·01及0·15±0·02,两组相比差异具有统计学意义(t=14·38,11·73,P均<0·05);实验组移植肾HO-1蛋白含量分别为(297±61)及(468±51)ng/g,而对照组分别为(98±30)及(155±31)ng/g,两组相比差异具有统计学意义(t=8·27,14·83,P均<0·05)。与对照组相比实验组移植肾病理改变明显减轻(P<0·05),血肌酐水平明显降低(t=8·41,P<0·05)。结论腺病毒载体可成功介导HO-1基因对大鼠肾脏的转移,对自体移植肾缺血再灌注损伤具有保护作用。

Objective To investigate the protective effect of Heme oxygenase-1(HO-1)gene transfer on rat renal autograft against ischemia/reperfusion injury. Methods HO-1 recombinant adenovirus vectors were contructed and transduced into rat renal autograft by renal arterial perfusion. The renal autografts were transplanted orthotopically after store at 4℃ for 24 h, followed by contralateral native nephrectomy 5 d after transplantation. There were 25 rats in the control group. 5 h and 3 d after transplantation , reverse transcriptase polymerase chain reaction(RT-PCR) and immunohistochemistry were used to detect the expression of HO-1 gene; enzyme-labeled immunosorbent (ELISA) was used to measure HO-1 protein content in the homogenate of renal autograft. Results The intensity of HO-1mRNA expression at 3 h and 3 d after transplantation were 0.65±0.11,0.86±0.17 in the experimental group and 0.09±0.01, 0.15±0.02 in the control group respectively. The differences between the two groups were significant (t=14.38,11.73,P<0.05). HO-1 protein content at 3h and 3d after transplantation were significantly increased in the experimental group, as compared with the control group [(297±61)ng/g and (468±51)ng/g versus (98±30)ng/g and (155±31)ng/g; t=8.27,14.83,P<0.05]. HO-1 transduced autografts had less renal ischemic injury and lower serum creatinine level compared with control animals (P<0.05). Conclusion Adenoviral vector can successfully transduce rat kidneys with the HO-1cDNA, which can protect rat renal autografts from ischemia/reperfusion injury.