大鼠骨髓内皮祖细胞移植对单侧输尿管梗阻大鼠肾脏损伤的修复

Effect of endothelial progenitor cells on renal impaivement of unilateral ureteral obstruction rats

目的探讨大鼠骨髓内皮祖细胞移植对单侧输尿管梗阻大鼠肾脏的修复作用。方法应用密度梯度离心法分离骨髓单个核细胞,内皮祖细胞选择性培养液培养后移植到单侧输尿管梗阻大鼠。应用PAS、HE、Masson染色方法判断梗阻大鼠损伤和修复情况,应用免疫组织化学检测-αSMA和HIF-1α表达,应用Western blot检测HIF表达。结果各组间肝功能、肾功能无显著性差异;与假手术组比,UUO组肾脏病理积分显著增加,移植组肾脏病理积分显著低于UUO组(P<0.05);UUO组α-SMA在14、21d显著高于假手术组(P<0.05),移植组-αSMA在14、21d显著低于UUO组(P<0.05);假手术组仅有少量HIF表达,且主要表达在胞质,UUO组HIF表达随病程进展逐渐增加(P<0.05),移植组HIF表达显著低于UUO组(P<0.05)。结论EPCs移植改善了UUO大鼠梗阻肾脏的损伤、纤维化和低氧状况。

Objective To reaserch effect of endothelial progenitor cells on renal impairment of unilateral ureteral obstruction （UUO） rats. Methods Mononuclear cells（MNCs） in rat bone marrow were isolated by density gradient centrifugation and were cultured according to previously described techniques, then transplanted by direct renal artery injection. Histopathological changes in kidney slices and severity were judged by interstitial lesion scoring by HE, PAS, and Masson methods. The expression of α-SMA and HIF-1α was detected by immunohistoehemistry method. The protein expression of HIF-1α was assessed by Western blot method. Results Hepatic function and renal function did not vary among transplantation group and UUO group at the same time. The interstitial fibrosis and tubular injury in transplantation group was lighter than that in UUO group. Compared with sham group, expression of alph-SMA was significantly higher in UUO group on 14,21d（P〈0.05）, but that in transplantation group was lower than that in UUO group（P〈0.05） respectively on 14,21d. There was trace amount of expression of HIF in sham group. But in UUO group that was significantly increased （P〈0.05） ,and on 21d reached at peak. And expression in transplantation group was lower than that in UUO group respectively on 7,14,21d（P〈0.05）. Conclusion EPCs transplantation improve renal interstitial damage, fibrosis and hypoxia of UUO rats.