目的研究后肾间充质干细胞(MMSC)在肾缺血再灌注损伤中的作用。方法将C57BIJ6J雄鼠分为正常组(n=5)和实验组(n=30)。正常组作为术前对照,实验组进行肾缺血再灌注损伤手术处理,根据股静脉注射物不同,再分为两个亚组:缺血再灌注组(注射生理盐水,10ml/kg)和细胞治疗组(注射含5×10^5个MMSC的生理盐水,10ml/kg)。实验组术后12、24、72h和正常组各取5只小鼠全血和肾脏,检测血清肌酐(SCr)水平,分析。肾组织病理染色结果和肾小管Paller评分,评估MMSC的作用。采用反转录一聚合酶链反应法检测肾组织的微小RNA.26a(miR.26a)水平,观察miR-26a在各组间的差异。结果(1)缺血再灌注组和细胞治疗组术后12、24、72h的SCr均高于正常组,且术后24h的SCr均高于术后12、72h,差异均有统计学意义(均P〈0.05);术后12、24、72h缺血再灌注组和细胞治疗组间SCr差异无统计学意义(均P〉0.05)。(2)缺血再灌注组和细胞治疗组术后12、24、72h的肾小管Paller氏评分均高于正常组,且24h评分〉72h评分〉12h评分(均P〈0.05);在术后24h,细胞治疗组的Paller氏评分低于缺血再灌注组[(57.2±6.3)比(70.8±14.8)分](P〈0.05);与正常组和细胞治疗组相比,缺血再灌注组术后24h可见明显的蛋白管型,肾小管萎缩、肿胀。(3)缺血再灌注组和细胞治疗组术后12、24h的肾组织miR-26a相对表达量均低于正常组(均P〈0.05);在术后24、72h,细胞治疗组肾组织miR-26a相对表达量分别是0.416±0.139、1.152±O.239,明显高于缺血再灌注组(0.244±0.067、0.855±0.038,均P〈0.05),且肾组织miR·26a的相对表达量与SCr呈负相关(r=-0.5,P〈0.05)。结论MMSC对肾缺血再灌注损伤有一定的修复作用,伴随着肾组织miR-26a表达量的增加。
Objective To study the effect of metanephric mesenchymal stem cells (MMSCs) after renal ischemia reperfusion injury (IRI). Methods C57BL/6J male mice were divided into control group (n = 5) and experimental group ( n = 30). The control group was given a sham operation, while in the experimental group, a model of renal IRI was established. The experimental group was further divided into two groups according to the material injected through the femoral vein: IRI group (injected with normal saline, 10 ml/kg ) and cell therapy group (injected with normal saline containing 5 x l0s MMSCs, 10 ml/kg). Samples of blood and kidney tissues were collected from five mice from each group at 12 h, 24 h and 72 h after IRI. The serum creatinine (SCr) level was detected, and the results of kidney tissue pathological staining and Paller score of renal tubules were analyzed to assess the effect of MMSCs after renal IRI. In addition, the expression of microRNA-26a (miR-26a) in kidney tissues was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and compared among the groups. Results ( 1 ) In both IRI group and cell therapy group, the levels of SCr at 12 h, 24 h and 72 h after operation were all significantly higher than those of the control group, besides, the level of SCr at 24 h was significantly higher than that at 12 h and 72 h (all P 〈0. 05). The levels of SCr at 12 h, 24 h and 72 h were of no significant differences between IRI group and cell therapy group ( all P 〉 0. 05 ). ( 2 ) Paller scores of renal tubules at 12 h, 24 h and 72 h in both IRI group and cell therapy group were significantly higher than those in the control group, and the scores at 24 h were significantly higher than that at 72 h, while the latter were in turn higher than the scores at 12 h (all P 〈 0. 05). In the cell therapy group, Paller score of renal tubules at 24 h (57.2 ± 6. 3 ) was significantly lower than that in IRI group (70. 8± 14. 8 ) ( P 〈 0. 05 ).