大鼠血红细胞活性氧在肾缺血再灌注损伤中的表达及作用研究

摘要

目的 探讨随着肾缺血再灌注(renal ischemia reperfusion injury,IRI)时间的延长,血液中活性氧(reactive oxygen species,ROS)的变化及其与肾功能、肾脏病理变化的关系.方法 将雄性SD大鼠42只,随机分为正常组5只、假手术组(sham group,SG)5只和模型组32只,模型组下设IRI后1、6、12和24h4个时间点,每个时间点8只大鼠.通过阻断大鼠双侧肾脏血流制成IRI模型.采用全自动生化分析仪检测各组大鼠的肾功能,糖原染色(periodic Acid Schiff,PAS)了解肾脏病理损害,流式细胞仪检测红细胞ROS.结果 随着缺血再灌注后时间的延长,大鼠肌酐、尿素氮水平逐渐升高,红细胞中ROS荧光强度阳性率逐渐升高,肾小管出现不同程度的管腔扩张、变性与坏死,间质炎性细胞浸润、充血水肿等变化.相关性分析发现,红细胞ROS荧光强度阳性率与肾小管损伤评分、肌酐、尿素氮水平成正相关(r=0.735,P＜0.01;r=0.682,P＜0.01;r=0.622,P＜0.01).结论 IRI后大鼠血红细胞ROS荧光强度阳性率明显上升；血红细胞ROS的改变可能参与了IRI肾脏的损伤过程.%Objective To investigate the changes of reactive oxygen species (ROS) in the blood and its relationship with the change of renal function and renal pathological as the time extension after renal ischemia-reperfusion (IRI). Methods A total of 42 male Sprague-Dawley rats were randomly divided into normal group (n=5) ,sham group (n = 5) and IRI group (n=32). IRI group was divided into four subgroups with different time points: 1 hour, 6 hour, 12 hour and 24 hour, each time point contain 8 animals. IRI model was established by blocking bilateral renal blood flow of the rats. Renal function was detected by using automatic biochemical analyzer; the renal pathological damage was estimated with periodic acid Schiff (PAS); and the ROS of erythrocyte was detected by flow cytometry. Results With the extension of the time after ischemia-reperfusion injury rats creatinine, the level blood urea nitrogen (BUN) and the positive rate of ROS fluorescence intensity in erythrocyte is gradually increased. Tubular can appearing vary degrees of luminal expansion and degeneration and necrosis and interstitial inflammatory cell infiltration and congestion and edema and so on. Correlation analysis found that the positive rate of erythrocyte ROS fluorescence intensity was positively correlated with tubular injury score, creatinine and blood urea nitrogen level(r=0. 735, P<0. 01; r=0. 682, P<0. 01; r=0. 622, P< 0. 01). Conclusion The erythrocyte ROS fluorescent intensity positive rate of rat was significantly increased after IRI. The erythrocyte ROS may be involved in the process of kidney damage in IRI.