白细胞介素-32在大鼠肾脏缺血再灌注损伤中的作用及其机制

Effect of IL-32 on Renal Ischemia-reperfusion Injury and its Mechanism in Rats

目的:探讨白细胞介素-32(Interleukin,IL-32)在大鼠肾缺血再灌注(ischemia-reperfusion injury,IRI)损伤中的作用及其机制。方法:80只雄性SD大鼠随机分为4组:假手术组(S组,n=20)、缺血再灌注组(I/R组,n=20)、缺血前IgG抗体预处理组(IgG+I/R组,n=20)和缺血前IL-32抗体预处理组(Anti-IL-32+I/R组,n=20)。采用夹闭双侧肾蒂30min后恢复血供的方法制备肾脏缺血再灌注损伤模型,再灌注3h、6h、12h、24h分别处死大鼠收集血样和肾脏样本,自动生化分析仪测定血清中肌酐(Cr)、尿素氮(BUN)浓度,酶联免疫吸附试验(ELISA)检测大鼠血清IL-32、TNF-α、IL-1β的表达,化学比色法检测肾组织髓过氧化物酶(MPO)的活性。结果:与S组比较:1、血清Cr、BUN浓度,I/R、IgG+I/R组均显著升高(P<0.05),Anti-IL-32+I/R组无显著性差异(P>0.05);2、血清IL-32、TNF-α、IL-1β水平,I/R、IgG+I/R组均显著升高(P<0.05),Anti-IL-32+I/R组无显著性差异(P>0.05);3、肾组织MPO活性,I/R、IgG+I/R组显著增强(P<0.05),Anti-IL-32+I/R组无显著性差异(P>0.05)。结论:IL-32在肾脏缺血再灌注损伤后的大鼠血清中高表达,阻断IL-32能减少炎症因子释放、抑制中性粒细胞聚集,减轻肾功能损害。

Objective：To investigate the effect of IL-32on renal ischemia-reperfusion injury and its mechanism in rats.Methods：Eighty male SD rats were randomly divided into 4groups：Sham group（S group,n=20）,renal IRI group（I/R group,n=20）,IgG preconditioning before ischemia group（IgG＋I/R group,n=20）and Anti-IL-32preconditioning before ischemia group（Anti-IL-32＋I/R group,n=20）.Both renal pedicales of rats were clamped for 30minutes to make the animal models of renal ischemia-reperfusion injury,then removed the clamps.3h,6h,12h,24hafter renal reperfusion,the plasma and kidneys were collected for detecting the serum creatinine,BUN level by auto biochemical analysis,IL-32,TNF-αand IL-1βexpression in serum by ELISA assay,activity of myeloperoxidase in kidney tissues by chemical chromatometry.Results：The levels of serum Cr and BUN in I/R group and IgG＋I/R group were significantly higher than S group（P〈0.05）,but there is no significant difference between Anti-IL-32＋I/R group and S group（P〈0.05）.Similarly,The levels of IL-32,TNF-αand IL-1βin I/R and IgG＋I/R group were significantly higher than S group（P〈0.05）,but there is no significant difference between Anti-IL-32＋I/R group and S group（P〈0.05）.The activity of MPO reduced significantly（P〈0.05）.Conclusions：IL-32is highly expressed in serum and kidney on renal ischemia-reperfusion injury.Blockages of IL-32could decrease the level of inflammatory,inhibit the movement of neutrophils,and reduce the renal histopathologic damage.