MicroRNA-21对肾缺血再灌注损伤的调控作用及其机制的研究

发布时间：2019-05-27 02:42

【摘要】：目的： 探索microRNA-21在肾缺血再灌注损伤中作用机制,以期找到防治肾缺血再灌注损伤microRNA分子靶标。 方法： 采用微型血管夹,同时夹闭雄性BALB/C小鼠双侧肾蒂构建实验所需的肾缺血再灌注损伤模型，参照前期实验的结果和本次试验的目的将小鼠分为Pre-miR-21+IRI组（阻断肾蒂血管45分钟，，再灌注24h，于术前24小时和6小时分别按200ng/kg腹腔注射mir-21agomir）、PBS+IRI组（阻断肾蒂血管45分钟，再灌注24h于术前24小时和6小时分别腹腔注射等体积的PBS）、Pre-miR-21+shame组（仅打开腹腔不阻断肾蒂血管，于术前24小时和6小时分别按200ng/kg腹腔注射mir-21agomir）、PBS+shame组（仅打开腹腔不阻断肾蒂血管，于术前24小时和6小时分别腹腔注射等体积PBS），（n≥5）。四组小鼠缺血再灌注损伤后，收集小鼠血液检测血清肌酐、尿素氮判断肾功能。断颈法处死小鼠，从小鼠背部取一手术切口获取双侧模型肾脏。一侧肾脏行HE染色，光学显微镜下观察模型肾病理组织变化，另一侧肾脏检测模型肾中microRNA-21，PDCD4、PTEN基因的mRNA和caspase-3蛋白的表达。 结果： （1）Pre-miR-21+IRI、PBS+IRI组、Pre-miR-21+shame组和PBS+shame组组小鼠存活率分别为87%、75%、100%、100%。Pre-miR-21+IRI组中血清肌酐、尿素氮和肾组织病理评分明显低于PBS+IRI组，但上述两组均高于PBS+shame组和Pre-miR-21+shame（P＜0.05）；PBS+shame组与Pre-miR-21+shame组肾组织病理评分和血清肌酐、尿素氮无明显差异(P＞0.05)。 （2）四组小鼠中miR-21的表达存在显著差异，从高到低的排列顺序为Pre-miR-21+IRI、Pre-miR-21+shame组、PBS+IRI组、PBS+shame组(P＜0.05)；Pre-miR-21+IRI组中靶基因PDCD4、PTEN的mRNA表达量低于PBS+IRI组（P＜0.05）；PBS+IRI组中靶基因PDCD4、PTEN的mRNA表达量低于PBS+shame组（P＜0.05）；PBS+shame组与Pre-miR-21+shame组两组之间靶基因PDCD4、PTEN的mRNA表达量无明显差异(P＞0.05)。 （3）、Pre-miR-21+IRI组中cleaved caspase-3表达明显低于PBS+IRI组（P＜0.05），PBS+IRI组中cleaved caspase-3高于PBS+shame组和Pre-miR-21+shame组（P＜0.05），差异均有统计学意义。各组中未经激活的pro-caspase-3无明显差异，P＞0.05无统计学意义。 结论： （1）成功构建了雄性BALB/c小鼠肾脏中mir-21过表达模型和肾缺血再灌注损伤模型； （2）mir-21能对其靶基因PDCD4和PTEN进行负向调控； （3）在肾缺血再灌注损伤的环境中mir-21能通过对其靶基因PDCD4和PTEN的负向调控，抑制肾缺血再灌注损伤所造成凋亡，从而对肾脏起到一定的保护作用。
[Abstract]:Objective: to explore the mechanism of microRNA-21 in renal ischemia-reperfusion injury in order to find a molecular target for the prevention and treatment of renal ischemia-reperfusion injury. Methods: the model of renal ischemia-reperfusion injury was established by clamping bilateral renal pedicle of male BALB/C mice with microvascular clip. According to the results of the previous experiment and the purpose of this experiment, the mice were divided into Pre-miR-21 IRI group (blocking renal pedicle vessels for 45 minutes, reperfusing them for 24 hours, injecting mir-21agomir intraperitoneally according to 200ng/kg 24 hours and 6 hours before operation). PBS IRI group (blocking renal pedicle vessels for 45 minutes, 24 hours after reperfusion, 24 hours before operation and 6 hours before operation, respectively, intraabdominal injection of the same volume of PBS), Pre-miR-21 shame group (only open abdominal cavity did not block renal pedicle vessels, Mir-21agomir), PBS shame group was injected intraperitoneally with 200ng/kg 24 hours and 6 hours before operation (only open abdominal cavity did not block renal pedicle vessels, and PBS), (n 鈮

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