PLGF通过激活Rho/ROCK信号通路介导人脑微血管内皮细胞间紧密连接开放

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (8): 960-965.

PLGF通过激活Rho/ROCK信号通路介导人脑微血管内皮细胞间紧密连接开放

王淳¹,李波²,方文刚³,陈誉华⁴

1. 辽宁中医药大学
2. 中国医科大学基础医学院
3. 中国医科大学基础医学院发育生物学教研室
4. 中国医科大学

收稿日期:2012-10-23 修回日期:2012-11-27 出版日期:2013-08-05 发布日期:2013-07-18
通讯作者: 陈誉华 E-mail:yhchen@mail.cmu.edu.cn
基金资助:
胎盘生长因子（PLGF）介导小细胞肺癌细胞经血脑屏障转移入脑的机制研究

Placental growth factor induces brain microvascular endothelial cell tight junctions’opening by activation of Rho/ROCK signaling pathway

Received:2012-10-23 Revised:2012-11-27 Online:2013-08-05 Published:2013-07-18

摘要/Abstract

摘要： 目的探讨胎盘生长因子（PLGF）影响人脑血管内皮细胞（HBMEC）间通透性的分子机制。方法利用HBMEC构建体外血脑屏障模型，测定跨内皮细胞电阻抗（TEER）及检测HRP通过率，分析PLGF对血脑屏障完整性的影响；用Western blot法及免疫荧光法检测HBMEC间紧密连接相关蛋白质occludin和ZO-1的表达及分布，用蛋白激酶抑制剂干预和Western blot法检测Rho蛋白活性。结果 25 ng/ml PLGF 30 min显著降低体外血脑屏障模型TEER，使其通透性增高（P＜0.05）；增加S-occludin表达的同时降低了IS-occludin的表达，使ZO-1蛋白在细胞周边分布减少；此外，ROCK蛋白激酶抑制剂Y27632抑制了PLGF引起的体外血脑屏障透过性增高（P＜0.05），并激活细胞内Rho激酶活性。结论 PLGF通过激活HBMEC内Rho/ROCK信号通路，影响HBMEC间紧密连接完整性，进而改变血脑屏障的通透性。

关键词: 胎盘生长因子, 血脑屏障, 紧密连接, Rho/ROCK

Abstract: Objective To study the molecular mechanism of placental growth factor (PLGF) inducing brain vascular endothelial cell tight junctions’ opening. Methods Using human brain microvascular endothelial cells (HBMECs) to construct blood-brain barrier (BBB) modal in vitro, with which to study human recombinant PLGF protein inducing the change of electrical resistance (TEER) and permeability of the BBB. Moreover, the distribution and change of tight junction protein ZO-1 and occludin were observed by immumofluorescence and Western blotting methods to confirm the effects of PLGF on tight junction of BBB. Finally, detected the cell signals induced by PLGF on tight junction of BBB using protein inhibitors and Western blotting method.Results PLGF induced a decrease of TEER and an increased of HRP flux of BBB modal in vitro. Besides, PLGF increased soluble occludin and changed ZO-1 distribution, the inhibitor of ROCK (Y27632) blocked the HRP flux induced by PLGF, and RhoA-GTP was decteced in HBMEC after PLGF tratement. Conclusion PLGF induces tight junction’s opening of BBB in virto by activation Rho/ROCK signaling.

Key words: placental growth factor, blood-brain barrier, tight junction, Rho/ROCK

王淳李波方文刚陈誉华. PLGF通过激活Rho/ROCK信号通路介导人脑微血管内皮细胞间紧密连接开放[J]. 基础医学与临床, 2013, 33(8): 960-965.

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