RT-PCR of microRNAs derived from rats: Ischaemia, Sevoflurane preconditioning, Ischaemic preconditioning, Sham model

We compared sevoflurane preconditioning and ischaemic preconditioning effects against renal ischaemia reperfusion injury via microRNAs of the kidney that promote cell-survival pathways in rats. In our protocol, male Wistar rats were divided into four groups (Ischaemia, Sevoflurane preconditioning, Ischaemic preconditioning and Sham models; n=7 each) and all underwent right nephrectomy. Ischaemia model rats underwent 45-minute clamping of left kidney, followed by 4-hour reperfusion. Sevoflurane preconditioning involved exposure to 1 MAC sevoflurane for 15 minutes. Ischaemic preconditioning included 3 cycles of 2-minute clamping and 5-minute reperfusion. Renal biopsy samples were assessed postoperatively to comprehensively analyse renal mircoRNAs using RT-PCR and the predicted cell-survival pathways. Sevoflurane preconditioning and ischaemic preconditiong ameliorated ischaemia reperfusion injury, indicated by serum creatinine equally. The result of RT-PCR indicated that sevoflurane and ischaemic preconditioning affected different kinds of microRNAs related to the cell-survival pathways respectively.本研究以大鼠为实验对象，通过肾脏中可调控细胞存活通路的微小RNA（microRNAs），对比七氟烷预处理与缺血预处理对抗肾缺血再灌注损伤的效果。实验方案中，将雄性维斯塔尔大鼠随机分为四组：缺血组、七氟烷预处理组、缺血预处理组及假手术组，每组各7只；所有大鼠均接受右肾切除术。缺血组大鼠行左肾蒂阻断45分钟，随后予以4小时再灌注。七氟烷预处理组大鼠予1 MAC七氟烷吸入15分钟。缺血预处理组大鼠则接受3次循环的左肾蒂阻断2分钟、再灌注5分钟操作。术后获取肾脏活检样本，通过逆转录聚合酶链反应（RT-PCR）全面分析肾脏微小RNA及其预测调控的细胞存活通路。七氟烷预处理与缺血预处理均能改善肾缺血再灌注损伤，二者的血清肌酐改善效果相当。逆转录聚合酶链反应结果显示，七氟烷预处理与缺血预处理分别调控了不同类型的细胞存活通路相关微小RNA。