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RhoA在内皮-单核细胞激活多肽-Ⅱ增强血肿瘤屏障通透性中的作用
引用本文:李振,刘云会,薛一雪,刘丽波,王萍. RhoA在内皮-单核细胞激活多肽-Ⅱ增强血肿瘤屏障通透性中的作用[J]. 解剖科学进展, 2014, 0(1): 68-72
作者姓名:李振  刘云会  薛一雪  刘丽波  王萍
作者单位:[1]中国医科大学附属盛京医院神经外科,辽宁沈阳110004 [2]中国医科大学基础医学院神经生物教研室,辽宁沈阳110001
基金项目:国家自然科学基金项目(No.81172197,81171131.30973079);高等学校博士学科点专项科研基金(No.20092104110015,20102104110009);沈阳市科学技术计划项目计划(No.F10-205-1-37,F10-205-1-22)
摘    要:
目的探索信号分子RhoA在内皮-单核细胞激活多肽-Ⅱ(endothelial monocyte activating polypeptide-Ⅱ,EMAP-H)增强血肿瘤屏障(blood-tumor barrier,BTB)通透性中的作用。方法采集出生3~5d的Wistar胎鼠的大脑皮质,应用酶消化法及葡聚糖离心法获得脑微血管段后,接种于培养皿中进行脑微血管内皮细胞(brain microvascular endothelial cells,BMECs)原代培养;将BMECs与C6脑胶质瘤细胞共培养,构建体外BTB模型;共培养后的BMECs随机分成3组(每组6例):对照组、EMAP-Ⅱ组和C3 exoenzyme+EMAP-Ⅱ组。测定跨内皮阻抗值和辣根过氧化物酶流量评估各组BTB通透性变化情况;Western blot法检测BMECs上紧密连接相关蛋白ZO-1的表达水平;免疫荧光法检测ZO-1和细胞骨架蛋白F-actin在BMECs上的分布与表达。结果与对照组相比较,EMAP-Ⅱ组BTB通透性显著增高,ZO-1和F-actin在BMECs上的表达水平显著降低,应力纤维形成明显增多;EMAP-Ⅱ的上述作用受到RhoA抑制剂C3 exoenzyme预处理的显著抑制。结论信号分子RhoA在EMAP-Ⅱ增强BTB通透性中发挥着重要的作用。

关 键 词:内皮-单核细胞激活多肽-Ⅱ  血肿瘤屏障  通透性  RhoA  Wistar胎鼠  大鼠

Role of signaling molecule RhoA in the endothelial monocyte-activating polypeptide-II-induced enhancement in blood-tumor barrier permeability
LI Zhen,LIU Yun-hui,XUEYt-xue,LIU Li-bo,WANG Ping. Role of signaling molecule RhoA in the endothelial monocyte-activating polypeptide-II-induced enhancement in blood-tumor barrier permeability[J]. Progress of Anatomical Sciences, 2014, 0(1): 68-72
Authors:LI Zhen  LIU Yun-hui  XUEYt-xue  LIU Li-bo  WANG Ping
Affiliation:1. Department of Neurosurgery of Shengjing Hospital, and 2. Department of Neurobiology of College of Basic Medicine, China Medical University, Shenyang 110001, China )
Abstract:
Objective To investigate the role of RhoA in the endothelial monocyte-activating polypeptide-II (EMAP-II)-induced enhancement in the blood-tumor barrier(BTB) permeability. Methods Relatively pure cerebral microvessel fragments were obtained from the cortex of 3-5 days old Wistar rats by using careful dissection, enzyme digestion and dextran centrifugation. Then, these fragments were seeded on dishes and cultured primarily. BTB models were constructed by co-cultivation of rat brain microvascular endothelial cells (BMECs) with C6 glioma cells in vitro. Confluent monolayers of co-cultured BMECs were divided randomly into 3 groups (n=6 for each): control, EMAP-II and C3 exoenzyme + EMAP-II groups. Transendothelial electric resistance values and horseradish peroxidase flux were measured to evaluate changes in the BTB permeability. The expression level of tight junction-related protein ZO-1 on BMECs was measured by western blot assay. Immunofluorescence was used to identify the expression and distribution of ZO-1 and actin-cytoskeleton protein F-actin on BMECs. Restdts Compared with control group, the BTB permeability of EMAP-II group was increased significantly, the expression levels of ZO-1 and F-actin on BMECs were significantly decreased, accompanied with marked increase in the stress fibers formation. These above-mentioned effects of EMAP-II were significantly inhibited by pretreatment with C3 exoenzyme, the inhibitor of RhoA. Conclusion Signaling molecule RhoA plays an important role in EMAP-II-induced increase in the BTB permeability.
Keywords:endothelial monocyte-activating polypeptide-II  blood-tumor barrier  permeability  RhoA  Wistar rat  rat
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