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缺血再灌注损伤大鼠肾脏miRNA表达谱的筛查与分析
查宜凤1,王嘉锋1,李和文2,边琪2,赖学莉2,于光2*
0
(1. 第二军医大学长海医院麻醉科,上海 200433;
2. 第二军医大学长海医院肾内科,上海 200433
*通信作者)
摘要:
目的 筛查可能与肾脏缺血再灌注损伤病理生理过程相关的miRNA。方法 利用SD大鼠构建肾脏缺血再灌注损伤模型,12 h后心脏穿刺取血检测血尿素氮与肌酐水平,并利用miRNA芯片检测肾脏miRNA表达谱的变化,最后通过生物信息学检索的手段初步探讨差异表达miRNA可能的作用靶点。结果 肾脏缺血再灌注12 h后血清尿素氮与肌酐水平显著升高,芯片检测结果显示肾脏有36条miRNA出现差异表达,其中差异在2倍以上的miRNA有15条。实时定量PCR验证的结果与芯片基本相符,表达上调的miRNA包括miR-290、miR-894、miR-292-5p、miR-327、miR-374、miR-98、miR-352、miR-132、miR-146b和miR-196a,下调的miRNA包括miR-145、miR-329、miR-375、miR-140*和miR-29a。生物信息学检索显示,这些miRNA可能与炎症反应、细胞死亡与增殖、血管再生和纤维化有关。结论 肾脏缺血再灌注损伤时多条miRNA表达发生了改变,其可能通过调控炎症反应、细胞死亡与增殖、血管再生和纤维化影响肾脏损伤的发生与发展,但具体机制有待进一步研究证实。
关键词:  微RNAs  再灌注损伤  急性肾损伤  炎症
DOI:10.3724/SP.J.1008.2014.00465
投稿时间:2013-11-20修订日期:2014-01-16
基金项目:国家自然科学基金(81372038),上海市卫生局局级课题(2009115).
miRNA expression profile in rat kidney during renal ischemia/reperfusion injury:screening and analysis
ZHA Yi-feng1,WANG Jia-feng1,LI He-wen2,BIAN Qi2,LAI Xue-li2,YU Guang2*
(1. Department of Anesthesiology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Nephrology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding authors.)
Abstract:
Objective To screen for miRNAs potentially involved in the pathogenesis of renal ischemia/reperfusion injury. Methods Renal ischemia reperfusion injury model was established with SD rats. Twelve hours after reperfusion, blood urea nitrogen and serum creatinine were determined and miRNA expression in the kidney was detected using miRNA microarray. Bioinformatics methods were used for a preliminary analysis of potential targets for differentially expressed miRNAs. Results Blood urea nitrogen and serum creatinine were both elevated 12 h after reperfusion. miRNA microassay showed 36 aberrantly expressed miRNAs, with 15 miRNAs having an expression level higher than 2 folds. Results of real-time PCR were generally in accordance with the microarray results. The elevated miRNAs included miR-290, miR-894, miR-292-5p, miR-327, miR-374, miR-98, miR-352, miR-132, miR-146b and miR-196a; and the down-regulated miRNAs included miR-145, miR-329, miR-375, miR-140* and miR-29a. Bioinformatics showed that these miRNAs were related to inflammation, cell death and proliferation, angiogensis and fibrosis. Conclusion Several miRNAs are aberrantly expressed during renal ischemia/reperfusion injury, which may influence renal injury through regulating inflammation, cell death and proliferation, angiogensis and fibrosis, but the exact mechanism remains to be further investigated.
Key words:  microRNAs  reperfusion injury  acute kidney injury  inflammation