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.

本研究旨在对比肾缺血再灌注损伤中，由肾脏微RNA介导的细胞存活通路在吸入性氟烷预处理与缺血预处理中的作用。在实验设计中，将雄性Wistar大鼠分为四组（缺血组、氟烷预处理组、缺血预处理组和假手术组；每组7只），所有大鼠均接受右侧肾切除术。缺血模型大鼠的左侧肾脏进行45分钟的夹闭，随后进行4小时的再灌注。氟烷预处理组大鼠暴露于1 MAC氟烷15分钟。缺血预处理组包括3个循环的2分钟夹闭和5分钟再灌注。术后对肾脏活检样本进行评估，以全面分析肾源微RNA，并采用RT-PCR技术预测细胞存活通路。结果显示，氟烷预处理与缺血预处理均能改善缺血再灌注损伤，血清肌酐水平得到同等程度的改善。RT-PCR的结果显示，氟烷预处理和缺血预处理分别影响了与细胞存活通路相关的不同微RNA。