RhoA在缓激肽选择性开放血肿瘤屏障中的作用  

Study of the effect of RhoA in bradykinin selectively increaseing the permeability of blood-tumor barrier

作　　者：马腾[1] 刘啸白[2] 薛一雪[1] 

机构地区：[1]中国医科大学基础医学院神经生物学教研室,辽宁沈阳110001 [2]中国医科大学第96期临床医学7年制,辽宁沈阳110001

出　　处：《中国药理学通报》2011年第2期187-191,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30872656,30670723,30973079);高等学校博士学科点专项科研基金(No20092104110015);沈阳市科学技术项目计划(No1072033-1-00,1081266-9-00)

摘　　要：目的探讨RhoA是否介导缓激肽开放血肿瘤屏障。方法应用RhoA的特异性抑制剂C3胞外酶(C3 ex-oenzyme)预处理体外血肿瘤屏障模型的大鼠脑微血管内皮细胞后,测量跨内皮阻抗值(TEER),辣根过氧化物酶(HRP)渗漏量,分析血肿瘤屏障通透性的改变;Western blot法检测紧密连接相关蛋白claudin-5可溶性片段向不溶性片段的转变;免疫荧光法和细胞荧光标记法观察体外血肿瘤屏障模型的大鼠脑微血管内皮细胞紧密连接相关蛋白claudin-5和丝状肌动蛋白结构和分布的改变。结果 C3 exoenzyme抑制缓激肽诱导TEER值的降低,HRP流量升高;C3 ex-oenzyme抑制claudin-5可溶性片段向不溶性片段的转变;C3exoenzyme抑制claudin-5由细胞膜向细胞质重新分布,抑制丝状肌动蛋白由细胞膜边缘向细胞中央区分布,分布于细胞边缘的F-actin明显增加,应力纤维形成明显减少。结论 RhoA介导缓激肽开放血肿瘤屏障。Aim To investigate the role of RhoA in bradykinin（BK）-induced the opening of blood-tumor barrier（BTB）.Methods Rat brain microvascular endothelail cells（RBMECs）of BTB model in vitro were pretreated with a specific RhoA inhibitor,C3 exoenzyme and then treated with bradykinin（BK）for 15 min.HRP flux and TEER assays revealed BTB permeability.An shift in claudin-5 distribution from insoluble to soluble fractions was observed by Western blot.The stress fiber formation and distribution of F-actin and claudin-5 in RBMECs of BTB model in vitro were assessed by fluorescence microscopy.Results C3 exoenzyme could partially inhibit endothelial leakage and restore normal TEER values in RBMECs of BTB model in vitro.An obvious shift in claudin-5 distribution from insoluble to soluble fractions was prevented by C3 exoenzyme.C3 exoenzyme inhibited BK-induced relocation of claudin-5 from cell membrane into the cytoplasm as well as stress fiber formation in RBMECs of BTB model in vitro.Conclusion RhoA plays a key role in BK-induced opening of BTB.

关 键 词：RHOA 缓激肽 血肿瘤屏障 紧密连接 CLAUDIN-5 肌动蛋白重排 

分 类 号：R-322[医药卫生]