摘要
目的研究Rho associated kinase(ROCK)特异性抑制剂Y-27632是否抑制缓激肽开放血肿瘤屏障。方法应用EVOM测定仪测定跨内皮阻抗值,分析血肿瘤屏障的通透性;应用辣根过氧化物酶渗漏实验分析血肿瘤屏障的通透性;应用免疫荧光方法观察原代大鼠脑微血管内皮细胞丝状肌动蛋白结构和分布的改变。结果BK作用15min时,跨内皮阻抗值最低,辣根过氧化物酶流量最高,血肿瘤屏障通透性最高;此时大鼠脑微血管内皮细胞边界的丝状肌动蛋白分布不连续,应力纤维形成增加。ROCK的特异性抑制剂Y-27632显著抑制了由缓激肽引起的血肿瘤屏障通透性升高和应力纤维的增加。结论Y-27632抑制缓激肽引起的血肿瘤屏障通透性升高,可能与丝状肌动蛋白结构和分布的改变和应力纤维的增加相关。
Objective To investigate the inhibitory effect of a specific Rho associated kinase（ROCK） inhibitor- Y-27632 on the opening of blood tumor barrier（BTB） by bradykinin（BK） and its possible mechanism. Methods The permeability of BTB was analysed by the value of transendothelial electric resistance （TEER） and horseradish peroxidase （HRP） flux, the formation and distribution of F-actin were observed by immunofluorescence. A relation between actin cytoskeleton rearrangement and the BK-induced opening of BTB was demonstrated by a specific ROCK inhibitor,Y-27632. Results TEER value of BTB was the lowest and the value of HRP flux of BTB reached a peak 15 min after injection of BK, at the same time, the distribution of F-actin in rat brain microvascular endothelial cells（RBMECs） was incontinuous and the number of stress fibers was increased compared with control. Y-27632 inhibited the increase of the permeability of BTB and the formation of stess fibers caused by BK. Conclusion The inhibition of Y-27632 on the increase of the permeability of BTB by BK might be related to the F-actin cytoskeleton rearrangement.
出处
《解剖科学进展》
CAS
2009年第4期400-402,407,共4页
Progress of Anatomical Sciences
基金
国家自然科学基金(No.30670723
No.30973079
No.30800451
No.30700861
No.30872656)
辽宁省高等学校科研项目计划(No.2008850)
辽宁省自然科学基金(No.2005212)
