【摘要】 目的探讨重组血红素氧合酶-1基因(Heme oxygenase-1,HO-1)腺相关病毒(rAVV-HO-1)对大鼠自体移植肾缺血再灌注损伤的保护作用。方法将48只雄性成年Lewis鼠随机分为两组。转染组(n=24)在供肾切取时应用rAVV-HO-1原位灌注;对照组(n=24)用磷酸盐缓冲溶液(PBS)原位灌注。所有供肾保存于4℃的UW液中24 h后行自体移植,同时切除对侧肾脏。留取术后3h、3d、7d的血清和尿标本以比较血肌酐、尿蛋白;取移植后3 h、3 d、7 d时的肾组织标本,以RT-PCR、免疫组化分析移植肾HO-1基因及蛋白的表达,间接法测定HO-1的活性。结果将rAVV-HO-1原位灌注后,转染组移植后3 h、3 d、7 d移植肾HO-1mRNA的表达强度分别为(0.55±0.13),(0.82±0.19)和(0.39±0.09),对照组移植后3 h、3 d、7 d的HO-1mRNA表达强度分别为(0.11±0.07)、(0.10±0.09)和(0.09±0.03),转染组在各时间点的HO-1 mRNA表达都明显高于对照组,差异均有统计学意义(P<0.05)。免疫组化结果显示转染组移植后3 h可见HO-1蛋白表达,而对照组无表达。3 d、7 d后转染组移植肾可见HO-1蛋白强阳性表达,而对照组仅可见HO-1蛋白微弱表达,两组具有显著差异。转染组移植后7 d血肌酐水平为(2.14±0.87)mg/dL,明显低于对照组(7.33±2.31)mg/dL,移植后7 d治疗组尿蛋白为(5.10±1.23)μg/mL,明显少于对照组(11.61±3.39)μg/mL,差异均有统计学意义(P<0.05)。移植后3 d、7 d治疗组胆红素浓度明显高于对照组(P<0.05)。结论用rAVV-HO-1转染大鼠移植肾可以诱导移植肾中HO-1的稳定表达,并对移植肾缺血再灌注损伤具有保护作用。更多还原

【Abstract】 【Objective】 To investigate the protective effect of heme oxygen-1(HO-1) gene transfer on rat autograft against ischemia/reperfusion injury.【Methods】rAVV/HO-1 was constructed.Donor Lewis rat kidneys were perfused with rAVV/HO-1(transplantation group,n =24) or PBS(control group,n =24),stored at 4℃ for 12 h,and transplanted orthotopically into Lewis rat recipients,followed by contralateral native nephrectomy.Post-transplant serum creatinine,urine protein,HO-1 mRNA and protein expression,and HO enzymatic activity were analyzed with corresponding methods at 3 h,3 d and 7 d after transplantation.【Results】In the gene transfer group HO-1 mRNA expression at 3 h,3 d and 7 d after transplantation was(0.55±0.13),(0.82±0.19) and(0.39±0.09),which was all significantly higher than that in the control group [(0.11 ±0.07),(0.10 ±0.09) and(0.09 ±0.03),separately,P <0.05).HO-1 protein could be detected in the gene transfer group at 3 h while no expression of HO-1protein was detected in the control group(P <0.05).HO-1 protein expression was significantly increased at 3 d and7 d after transplantation while the control group only exhibited weak expression.The serum creatinine level at 7 dafter transplantation was(2.14±0.87) mg/dL in the gene transfer group and(7.33±2.31) mg/dL in the control group,the difference of both groups was significant(P <0.05).The urine protein at 7 d was higher in control group than thatin the experimental group [(5.10±1.23) μg/mL vs(11.61±3.39) μg/mL,P <0.05)].Concomitantly,HO enzymaticactivity was increased at 3 d and 7 d after transplantation in the experimental group compared with that in thecontrol group(P <0.05).【Conclusion】rAVV/HO-1 can be successfully transferred into donor kidneys,which canprotect rat renal autografts from ischemia / reperfusion injury.更多还原