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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 autologous renal allograft ischemia-reperfusion injury in rats. Methods The expression vector of HO-1 adenovirus was constructed and transplanted into renal tissue of 26 rats (experimental group) after renal artery perfusion. After 4 days of storage, autotransplantation was performed and the contralateral kidney was excised 5 days after transplantation. Twenty-five rats were Control. At 3 and 3 days after transplantation, the expression of HO-1 gene and protein in transplanted kidney was detected by RT-PCR and immunohistochemistry. The renal tissue homogenate was determined by enzyme linked immunosorbent assay (ELISA) The content of HO-1 protein in pulp (expressed as absorbance value). Results At 3 and 3 days after transplantation, the expression of HO-1mRNA in experimental group was 0.65 ± 0.11 and 0.86 ± 0.17, respectively, while the control group was 0 · 09 ± 0 · 01 and 0 · 15 ± 0.02, there was significant difference between the two groups (t = 14.38, 11.73, P <0.05). The content of HO-1 in experimental group was (297 ± 61) And (468 ± 51) ng / g, respectively, while the control group was (98 ± 30) and (155 ± 31) ng / g respectively. There was significant difference between the two groups (t = 8.27,14.83, P <0.05). Compared with the control group, the pathological changes of renal transplantation in experimental group were significantly reduced (P <0.05), and the level of serum creatinine was significantly decreased (t = 8.41, P <0.05). Conclusions Adenovirus vector can successfully mediate the transfer of HO-1 gene to rat kidney and protect the autologous transplantation-induced renal ischemia-reperfusion injury.