缺血后处理对大鼠肾脏miRNAs表达谱的影响  

Expression Profiling of miRNAs in Rat Renal Ischemic Post-Conditioning

作　　者：王春阳[1] 叶冬波 倪少滨[1] 陈起引[1] 赵忠山[1] 麻立[1] 

机构地区：[1]哈尔滨医科大学附属第一医院,黑龙江哈尔滨150001

出　　处：《现代生物医学进展》2015年第28期5408-5413,5458,共7页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅科学技术研究面上项目(12521198);黑龙江省卫生厅科研课题(2012-528)

摘　　要：目的:观察缺血后处理对Wistar大鼠肾组织miRNAs表达的影响。方法:将20只Wistar大鼠随机分为缺血再灌注组(I/R组)和缺血后处理组(IPO组),每组10只。I/R组行剖腹手术,分离双肾动脉后切除右肾,夹闭左肾动脉阻断血供,45 min后恢复;IPo C组行剖腹手术,分离左肾动脉后将其夹闭,在阻断血液供应45 min后恢复血供时采取10 s再灌、10 s停灌处理,反复5个周期。在所有实验完成6小时后,处死大鼠切除左肾,分别提取两组肾组织中的总RNA和miRNA,利用miRNA微阵列对其进行杂交检测,通过芯片扫描和数据聚类分析,获取两组大鼠肾脏组织miRNAs表达谱,并进行q RT-PCR验证,筛选差异表达的miRNAs。结果:通过q RT-PCR验证共筛选出7种表达差异较大的miRNAs,其中下调的三种,分别是miR-27a,miR-665,miR-let7f,上调的四种,分别为miR-532-3p,miR-205,miR-122,miR-291b。结论:差异表达miRNAs可能参与缺血后处理减弱缺血再灌注损伤的作用机制中。Objective： To observe the expression profiling of miRNAs in Wistar rats renal ischemic postconditioning. Methods：According to the principle of random, the 20 Wistar rats were divided into ischemia reperfusion group（group I/R） and ischemic postconditioning group（IPO group）. The I/R group underwent laparotomy, removed the right kidney after separate bilateral renal pedicle,45 minutes after occlusion of the left renal artery to restore blood supply; the IPO group underwent laparotomy, separation of the left renal artery and then clipping it, non-invasive blood vessel occlusion 45 min after recovery of blood supply to 10 s reperfusion, 10 s stopping irrigation, 5 cycles after processing clamp. After the completion of the experiment, the kidney tissues of the I/R and IPO of the two groups of rats were removed. The total RNA and miRNA of two kidney tissues were extracted. In miRNA microarray hybridization,the chip scanning and data clustering analysis was performed, and got two groups of rat kidney miRNAs expression. Real time quantitative RT-PCR was performed to confirm the results obtained by microarray analysis. Results： By q RT-PCR validation, 7 miRNAs with great differential expression were selected, including three downregulated, respectively miR-27 a, miR-665, miR-let7 f, four up-regulated, respectively miR-532-3p, miR-205, miR-122, miR-291 b. Conclusions： Differential expression of miRNAs may be involved in the mechanism of ischemic postconditioning which attenuated ischemia reperfusion damage.

关 键 词：缺血后处理 肾脏 MIRNAS 基因芯片 

分 类 号：Q95-3[生物学—动物学] R692[医药卫生—泌尿科学]