| 摘要: |
| 目的 探讨RNA干扰排斥性导向分子a (repulsive guidance molecule a,RGMa)对大鼠脑缺血再灌注损伤后大脑皮质血管再生的作用及其可能的机制。方法 大鼠随机分为假手术组(S组)、脑缺血再灌注损伤组(I/R组)、脑缺血再灌注损伤+RGMa特异性RNA干扰重组腺病毒干预组(I/R+rAd-shRGMa组)以及脑缺血再灌注损伤+空载体重组腺病毒注射组(I/R+rAd-HK组)。大鼠在脑缺血前接受腺病毒注射,然后采用线栓法制备大鼠大脑中动脉缺血再灌注损伤模型。再灌注后2 d免疫荧光双标法定位RGMa及其受体Neogenin在血管内皮细胞上的表达。腺病毒注射后7 d,免疫组化标记CD31+的血管内皮细胞,计数大脑皮质缺血周围区微血管数;蛋白质印迹实验检测血管内皮生长因子a(VEGFa)蛋白表达水平;评估神经功能缺损。结果 在缺血再灌注损伤后,RGMa及其受体Neogenin在CD31+细胞上均有表达。RNA干扰抑制RGMa表达后,微血管数量增多,VEGFa蛋白表达水平升高,神经功能缺损减轻。结论 RNA干扰RGMa表达可促进缺血再灌注损伤后缺血皮质周围区血管再生,该过程可能与脑组织VEGFa表达水平上调有关。 |
| 关键词: 排斥性导向分子a 脑缺血 再灌注损伤 血管再生 |
| DOI：10.16781/j.0258-879x.2016.04.0441 |
| 投稿时间：2015-11-03修订日期：2015-11-30 |
| 基金项目:国家自然科学基金面上项目(81271307). |
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| Effect of RGMa-targeted RNA interference on angiogenesis in rat cerebral cortex after focal cerebral ischemia/reperfusion injury |
| WANG Yu,QIN Xin-yue*,ZHANG Rong-rong,XING Xiang-feng,XIE Fei |
(Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Neurology, Chongqing 400016, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of RNA interference targeting repulsive guidance molecule a (RGMa) on angiogenesis in rat cerebral cortex after focal cerebral ischemia/reperfusion (I/R) injury and its potential mechanism. Methods The experimental animals were randomly divided into sham operated group(S), cerebral I/R group, I/R plus RGMa-specific recombinant adenovirus (I/R+rAd-shRGMa) and I/R plus empty carrier recombinant adenovirus group (I/R+rAd-HK). Adenovirus were injected into the right cortex and hippocampus in rats before undergoing I/R surgery. The transient middle cerebral occlusion (tMCAO) model was induced by ligation with nylon monofilament. Location of RGMa and Neogenin was assessed by Laser confocal microscope 2 days after tMCAO. Seven days after adenovirus injection, the expression of CD31 was detected by immunohistochemistry and the expression of vascular endothelial growth factor a (VEGFa) was detected by Western blotting analysis. Neurological score was evaluated before animals were sacrificed. Results Both RGMa and Neogenin were expressed in CD31+ cells after focal cerebral I/R injury as shown by double staining. Suppression of RGMa via RNA interference increased the number of CD31+ cells and the expression of VEGFa, and improved the neurological deficits. Conclusion RGMa-targeted RNA interference can promote angiogenesis in peri-infarction cortex after cerebral I/R injury, which is partly related to the increase of VEGFa expression in the cerebral tissues. |
| Key words: repulsive guidance molecule a brain ischemia reperfusion injury angiogenesis |
