沉默排斥性导向分子A对脑缺血再灌注损伤大鼠血脑屏障的保护作用

doi:10.16098/j.issn.0529-1356.2019.03.001

解剖学报 ›› 2019, Vol. 50 ›› Issue (3): 269-274.doi: 10.16098/j.issn.0529-1356.2019.03.001

• 神经生物学 • 下一篇

沉默排斥性导向分子A对脑缺血再灌注损伤大鼠血脑屏障的保护作用

李敏¹ 伏红霞¹ 张艳梅¹ 钟义良² 秦新月^2*

1.内蒙古自治区人民医院神经内科，呼和浩特 010017; 2.重庆医科大学附属第一医院神经内科，重庆 400016

收稿日期:2018-09-18 修回日期:2018-11-03 出版日期:2019-06-06 发布日期:2019-06-06
通讯作者: 秦新月 E-mail:2711382863@qq.com
基金资助:
RGMa对缺血再灌注脑损伤后血脑屏障紧密连接蛋白的影响及机制

Silencing repulsive guidance molecule A showing protective effects on blood brain barrier after the injury of cerebral ischemia and reperfusion in rats

LI Min¹ FU Hong-xia¹ ZHANG Yan-mei¹ ZHONG Yi-liang² QIN Xin-yue ^2*

1.Department of Neurology, Inner Mongolia Autonomous Region People’s Hospital, Hohhot 010017, China;2.Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China

Received:2018-09-18 Revised:2018-11-03 Online:2019-06-06 Published:2019-06-06
Contact: QIN Xin-yue E-mail:2711382863@qq.com
Supported by:
EFFECTS AND MECHANISM OF RGMA ON BLOOD BRAIN BARRIER IN RATS WITH ISCHEMIA REPERFUSION

摘要/Abstract

摘要：

目的观察沉默排斥性导向分子A（RGMa）对脑缺血再灌注损伤大鼠血脑屏障及紧密连接蛋白的影响。方法雄性成年 SD 大鼠立体定位侧脑室注射腺病毒后建立大脑中动脉闭塞(MCAO) /再灌注（I/R）模型。将成年雄性SD大鼠40只，分为空白对照组（sh-con）和RGMa干扰组（sh-RGMa），分别在注射后1 d和3d观察腺病毒对RGMa的影响。将其余120只SD大鼠随机分为假手术组（sham）、缺血再灌注组（I/R）、空病毒组（I/R+sh-con）及病毒组（I/R+sh-RGMa）；I/R 72 h 后采用神经功能缺损评分评估各组大鼠神经功能恢复情况; 采用2，3，5-氯化三苯基四氮唑(TTC)染色测量脑梗死体积；采用静脉注射伊文思蓝观察血脑屏障通透性的改变；应用Western blotting和免疫组织化学染色检测RGMa的变化；应用Western blotting检测血脑屏障完整性相关紧密连接蛋白5（claudin-5）、基质金属蛋白酶9（MMP-9）和闭锁小带蛋白1（ZO-1）的表达。结果缺血再灌注后 72 h，I/R组较 sham 组神经功能评分降低，沉默RGMa能改善血脑屏障通透性，减少脑梗死体积；可下调MMP-9及上调claudin-5和ZO-1的表达。结论沉默RGMa对脑缺血再灌注损伤大鼠的血脑屏障有保护作用。

关键词: 排斥性导向分子A, 缺血再灌注损伤, 血脑屏障, 免疫印迹法, 大鼠

Abstract:

Objective To investigate whether silencing repulsive guidance molecule A (RGMa) shows protective effects on blood brain barrier (BBB) and tight junction protein after the injury of cerebral ischemia and reperfusion (I/R) in rats. Methods Middle cerebral artery occlusion (MCAO) /reperfusion was employed to establish the models in the male adult rats. Fourty male Sprague-Dawley rats were divided into blank control group (sh-con) and RGMa interference group (sh-RGMa). The effects of adenovirus on RGMa were observed at day 1 and day 3 after injection. The remaining 120 SD rats were randomly divided into sham group, I/R group, I/R+sh-con group and I/R+sh-RGMa group. After reperfusion for 72 hours, the neurological recovery was evaluated in rats by neurological deficit score，the infarcted volume was measured by 2,3,5-triphenyl tetrazolium chloride (TTC) staining and the permeability of BBB was performed through evans blue by tail vein injection. The expression of RGMa was detected by Western blotting and immunohistochemistry assay. The expression of claudin-5, matrix metalloprotein 9 (MMP-9）and zonula occludin 1（ZO-1）were detected by Western blotting. Results Silencing RGMa could improve the permeability of BBB, the infarcted volume, down-regulate the expression of MMP-9, and up-regulate the expression of claudin-5 and ZO-1. Conclusion Silencing RGMa shows protective effects on BBB after I/R injury in rats.

Key words: Repulsive guidance molecule A, Ischemia and reperfusion injury, Blood brain barrier, Western blotting, Rat

李敏伏红霞张艳梅钟义良秦新月 . 沉默排斥性导向分子A对脑缺血再灌注损伤大鼠血脑屏障的保护作用[J]. 解剖学报, 2019, 50(3): 269-274.

LI Min FU Hong-xia ZHANG Yan-mei ZHONG Yi-liang QIN Xin-yue. Silencing repulsive guidance molecule A showing protective effects on blood brain barrier after the injury of cerebral ischemia and reperfusion in rats[J]. Acta Anatomica Sinica, 2019, 50(3): 269-274.

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沉默排斥性导向分子A对脑缺血再灌注损伤大鼠血脑屏障的保护作用

Silencing repulsive guidance molecule A showing protective effects on blood brain barrier after the injury of cerebral ischemia and reperfusion in rats

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