重组人促红细胞生成素促进小鼠缺血再灌注损伤肾脏HO-1表达

Recombinant human erythropoietin promotes expression of HO-1 mRNA after renal ischemia reperfusion injury in mice

目的观察重组人促红细胞生成素(rhEPO)对小鼠缺血再灌注损伤(IR)肾脏血红素加氧酶-1(HO-1)mRNA表达的影响,探讨rhEPO对IR的保护作用。方法 90只雄性C57BL/6小鼠随机分为3组:假手术组(n=30)、IR组(n=30)及rhEPO干预组(n=30),分别于再灌注1、2、3、6、24、48h处死小鼠取相应标本,处死前留血标本进行肾功能检测(血肌酐),采用PAS染色评估肾组织形态学改变、TUNEL染色分析并比较各组凋亡细胞数量的变化,Real-time PCR检测肾脏组织中HO-1、白细胞介素6(IL-6)mRNA表达。结果再灌注3、6、24h,rhEPO干预组HO-1mRNA表达高于IR组,差异有统计学意义(P<0.05);再灌注6、24、48hrhEPO干预组IL-6mRNA表达低于IR组,差异有统计学意义(P<0.05)。rhEPO干预组24h血肌酐含量与IR组相比降低(P<0.05);rhEPO干预组肾脏病变与IR组比较减轻。TUNEL染色示:与假手术组相比,IR组凋亡细胞数量增多(P<0.05),rhEPO处理后凋亡细胞数量少于IR组(P<0.05)。结论 rhEPO可能通过促进IR小鼠肾脏HO-1表达对抗缺血再灌注损伤,减轻炎症损伤,改善肾功能。

Objective To investigate the effects of recombinant human erythropoietin （rhEPO） on the expression of heme oxygenase-l（HO-1） mRNA after renal ischemia/reperfusion （IR） in mice. Methods Ninety male C57BL/6 mice were randomly divided into three groups, namely, the sham operation group （n=30）, renal IR group （n=30）, and rhEPO treatment group （n=30）. Mice were sacrificed at 1, 2, 3, 6, 24, and 48 h after renal reperfusion, and the renal function was evaluated by determining blood ereatinine. Histological damages were observed using a semi-quantitative histomorphological scoring system from 0 to 4. Cell apoptosis was analyzed by TUNEL staining in each group. HO 1 and IL-6 mRNA expression was examined by real-time PCR. Results Compared with renal IR group, the expression of HO-1 mRNA was significantly higher in rhEPO treatment group at 3, 6, and 24 h after reperfusion（P〈0.05）. The expression of IL-6 mRNA was significantly lower in the rhEPO treatment group at 6, 24, and 48 h after reperfusion（P〈0.05）. Serum creatinine level in the rhEPO treatment group was significantly lower than that in the renal IR group at 24 h after reperfusion（P〈0.05）. Compared with the renal IR group, renal histology injury was greatly attenuated by rhEPO in rhEPO treatment group. TUNEL staining analysis indicated that the apoptotic cells in the IR group were significantly more than those in the sham operation group（P〈0. 05）, and those in the rhEPO treatment group was significantly less than those in the IR group（P〈0.05）. Conclusion rhEPO can attenuate renal ischemia/reoerfusion iniurv in mice, orobablv through promoting the renal expression of HO-1 mRNA.