MicroRNA-29c靶向调控BIRC2在预电刺激小脑顶核后大鼠局灶性脑缺血再灌注损伤中的作用

发布时间：2018-04-18 06:46

本文选题：MicroRNA-29c + BIRC2　；参考：《广西医科大学》2014年硕士论文

【摘要】：目的探讨MicroRNA-29c（miR-29c）在预电刺激小脑顶核后大鼠局灶性脑缺血再灌注损伤中的保护机制。方法将60只Sprague Dawley雄性大鼠采用随机数字表法分为3组：（1）假手术组；（2）单纯造模组；（3）预电刺激组：即首先预电刺激小脑顶核1h，24h后再行右侧局灶性脑缺血，2h后再灌注。然后用TTC染色检测脑梗死体积比例；RT-qPCR检测miR-29c、BIRC2mRNA表达量；Western-blotting检测BIRC2蛋白表达量；TUNEL染色检测凋亡的细胞数；构建BIRC2荧光素酶报告基因载体，双荧光素酶报告基因检测系统检测荧光素酶的表达变化。结果（1）脑梗死体积比例测定结果：预电刺激组与单纯造模组相比，脑梗死体积比例减小30.2%（p0.01）。（2）PCR检测结果：假手术组、预电刺激组、单纯造模组：miR-29c的相对表达量分别为（1.0、0.343±0.009、0.955±0.212），预电刺激组较单纯造模组明显下降，下降约3倍（p0.01）；BIRC2mRNA的相对表达量分别为（1.0、2.404±0.330、1.168±0.247），预电刺激组较单纯造模组明显增高，增高2倍多（p0.01）。（3）BIRC2蛋白表达量检测结果：假手术组、预电刺激组、单纯造模组BIRC2蛋白的相对表达量分别为（0.5±0.364、1.407±0.270、0.939±0.227），预电刺激组较单纯造模组明显增加，，约增高1.5倍（p0.05）。（4）TUNEL凋亡细胞数检测结果：预电刺激组凋亡细胞数（22.37±1.27）较单纯造模组凋亡细胞数（35.9±1.93）明显减少，约减少37.68%（p0.01）。（5）双荧光素酶报告基因系统显示：转染野生型载体组的荧光素酶活性显著下调，而转染突变型载体组的荧光素酶活性变化不明显，说明miR-29c可特异性抑制包含有BIRC23’UTR野生型识别元件报告基因的表达（p0.01）。结论预电刺激小脑顶核对其后的脑缺血再灌注损伤有保护作用，其机制可能与miR-29c通过靶向调节BIRC2的表达、减轻局灶性脑缺血再灌注损伤有关。
[Abstract]:Objective to investigate the protective mechanism of microRNA-29cmmiR-29c in focal cerebral ischemia-reperfusion injury after prestimulation of cerebellar parietal nucleus in rats.Methods Sixty Sprague Dawley male rats were randomly divided into 3 groups: control group (n = 1) sham operation group (n = 2) model group (n = 2) prestimulation group (n = 3): first prestimulation of cerebellar fastigial nucleus for 24 h and then right focal cerebral ischemia for 2 h and then reperfusion.Then TTC staining was used to detect the proportion of cerebral infarction volume and RT-qPCR was used to detect the expression of miR-29cnBIRC2 mRNA. Western-blotting was used to detect the expression of BIRC2 protein. Tunel staining was used to detect the number of apoptotic cells, and the BIRC2 luciferase reporter gene vector was constructed.The expression of luciferase was detected by double luciferase reporter gene detection system.Results 1) the ratio of cerebral infarction volume was measured: compared with the control group, the ratio of cerebral infarction volume in the prestimulation group was smaller than that in the control group: sham operation group, preelectric stimulation group, sham operation group, preelectric stimulation group,The relative expression of miR-29c in the control group was 0.343 卤0.009 卤0.955 卤0.212g, respectively. The relative expression of BIRC2 mRNA in the prestimulation group was significantly lower than that in the control group, and the relative expression of BIRC2 mRNA in the prestimulation group was about 3 times lower than that in the control group. The relative expression of BIRC2 mRNA in the prestimulation group was significantly higher than that in the control group, and the relative expression of BIRC2 mRNA in the preelectric stimulation group was significantly higher than that in the control group, and the relative expression of miR-29c mRNA in the prestimulation group was significantly higher than that in the control group.The results showed that the relative expression of BIRC2 protein in sham operation group, preelectric stimulation group and simple model group was 0.5 卤0.364 卤0.270 卤0.939 卤0.227, respectively. The expression of BIRC2 protein in preelectric stimulation group was significantly higher than that in the simple model group, and the expression of BIRC2 protein was significantly higher in the prestimulation group than in the control group, and the relative expression of BIRC2 protein in the sham operation group, preelectric stimulation group and simple model group were 0.5 卤0.364 卤0.270 卤0.939 卤0.227, respectively.The number of apoptotic cells in preelectric stimulation group (22.37 卤1.27) was significantly lower than that in model group (35.9 卤1.93).The system of double luciferase report gene showed that the luciferase activity of the wild-type vector group decreased significantly, but the luciferase activity of the mutant vector group did not change significantly.These results suggest that miR-29c can specifically inhibit the expression of a reporter gene containing BIRC23'UTR wild type recognition element.Conclusion prestimulation of cerebellar fastigial nucleus has protective effect on cerebral ischemia-reperfusion injury, and its mechanism may be related to the regulation of BIRC2 expression by targeting by miR-29c and the reduction of focal cerebral ischemia-reperfusion injury.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3

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