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目的研究肾脏来源的间充质细胞(KDMSC)对大鼠肾脏急性缺血损伤的作用并探讨其作用机制。方法建立肾脏缺血再灌注模型,实验动物随机分为4组,每组10只:对照组、缺血模型组、培养基对照组、干细胞治疗组。贴壁法分离筛选获得间充质干细胞,并原代培养至第3代,采用CM-DiI荧光染料标记后,应用微量注射器将干细胞直接注射入缺血的肾脏部位,3 d后观察大鼠血浆肌酐,尿素氮含量;RT-PCR观察肾脏组织IL-6,TNF-α表达情况;免疫组织化学检测KDMSC存活及血管内皮生长因子(VEGF)表达。结果与模型组相比,干细胞组血浆肌酐(Cr)与尿素氮(BUN)水平明显下降(P<0.05);IL-6与TNF-α表达降低(P<0.05);免疫组织化学染色显示,3 d后干细胞仍然存活,部分细胞与VEGF阳性细胞共存。结论KDMSC移植能明显改善大鼠缺血再灌注大鼠肾功能,可能与抑制炎症因子释放有关。
Objective To investigate the effect of kidney-derived mesenchymal cells (KDMSC) on renal acute ischemic injury in rats and its possible mechanism. Methods Renal ischemia-reperfusion model was established. The experimental animals were randomly divided into 4 groups with 10 animals in each group: control group, ischemia model group, medium control group and stem cell treatment group. MSCs were isolated and screened by adherence method and were primary cultured to the third passage. After labeling with CM-DiI fluorescent dye, the stem cells were directly injected into the ischemic kidney by microinjector. After 3 days, the rat plasma Creatinine and urea nitrogen. The expression of IL-6 and TNF-α in renal tissues was detected by RT-PCR. The survival of KDMSCs and the expression of vascular endothelial growth factor (VEGF) were detected by immunohistochemistry. Results Compared with the model group, the levels of serum creatinine (Cr) and urea nitrogen (BUN) were significantly decreased (P <0.05) and the expressions of IL-6 and TNF-α were decreased in the stem cells group (P < After 3 days, the stem cells still survived, and some cells coexisted with VEGF positive cells. Conclusion KDMSC transplantation can significantly improve renal function in rats with ischemia-reperfusion injury, which may be related to the inhibition of the release of inflammatory cytokines.