人胎盘间充质干细胞条件培养基对急性缺血再灌注肾损伤的修复作用

The mechanism of mesenchymal stem cells conditional medium derived from human placenta repairing the acute kidney ischemia-reperfusion injury

目的观察尾静脉注射人胎盘间充质干细胞条件培养基(human placenta-mesenchymal stem cell conditional medium,MSCs-CM)对急性缺血再灌注肾损伤(acute renal ischemia reperfusion injury,IR)的修复作用。方法健康雄性SD大鼠(n=30)分为3组:空白对照组(sham组),模型组(IR组)和治疗组(IR+MSCs组),每组10只。制备大鼠缺血再灌注肾损伤模型,建模24h后以尾静脉注射MSCs条件培养基,并连续跟踪4d监测血清及尿液的肌酐含量,随后取肾脏组织行过碘酸雪夫反应(PAS)染色检查观察肾脏损伤及修复状况;经2',7'-二氯荧光黄双乙酸盐(DCFH-DA)法检测肾组织内活性氧含量;Western Blot法检测肾脏组织内血红素氧化酶1(heme oxygenase-1,HO-1)及磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶1(NAPDH quinineoxidoreductase-1,NQO-1)的相对表达水平。结果模型组大鼠的血、尿肌酐含量相对于空白正常组明显升高,出现肾损伤;而治疗组大鼠的血、尿肌酐含量与IR组相比有明显改善且与Sham组指标相似;PAS染色观察显示,模型组大鼠存在肾小管损伤,而治疗组大鼠肾小管损伤明显轻于模型组;Western blot分析显示治疗组细胞抗氧化标志物NQO-1和HO-1的蛋白表达水平显著高于模型组(P<0.05)。结论 MSCs-CM经尾静脉移植后,对缺血再灌注所致的肾损伤具有明显的修复作用,其修复肾脏损伤机制可能与抗氧化应激有关。

Objective To observe the protective effect of tail vein injection of human placenta mesenchymal stem cell conditional medium（MSCs-CM） repairing ischemia reperfusion injury（IRI）.Methods Totally 30 healthy SD male rats were randomly divided into 3 groups,sham group,renal ischemia reperfusion model group（IR group） and MSCs conditional medium injection group（IR ＋ MSCs group） with ten rats in each group.The rodent model of ischemia-reperfusion（IR） injury of kidney was established,MSCs conditional medium were administrated into caudal vein at 24 h after IR.The Cr in serum and urine with consecutive 4 days were detected,and then kidney specimens were performed by PAS staining after rats were sacrificed,DCFH-DA was used to detect the content of active oxygen in renal tissue,and the relative expression of HO-1 and NQO-1 were detected by Western Blot.Results Compared with IR group,the creatinine from serum and urine in MSCs treated group（IR ＋ MSCs group） were ameliorated apparently after MSCs-CM injection.The pathological histology results demonstrated that the morphological damage of the renal tubule was observed in IR group,and amelioration was observed in MSCs treated group（IR ＋ MSCs group）.PAS showed that rats in the sham group had renal tubular injury,which was more severe than that of the MSCs treated group（IR ＋ MSCs group）.Western Blot showed notably higher NAPDH quinone oxidoreductase 1（NQO1） and HO-1 protein expression,the two indicators of anti-oxidative capacity,in MSCs treated group（IR ＋ MSCs group） than in sham group.Conclusion MSCs-CM transplanted after caudal vein is helpful for the repair of acute kidney injury induced by ischemia reperfusion injury.The repair mechanism might relate to the anti-oxidative stress.