肌球蛋白轻链激酶在烧伤大鼠血清引起的内皮细胞骨架改变过程中的作用(英文)  

Myosin light-chain kinase contributes to short-term endothelial cell cytoskeletal alteration induced by serum from burned rats

作　　者：陈波[1] 郭晓华[1] 王述昀[1] 黄绪亮[1] 黄巧冰[1] 

机构地区：[1]第一军医大学病生教研室,广东广州510515

出　　处：《第一军医大学学报》2004年第5期481-484,488,共5页Journal of First Military Medical University

基　　金：Sponsored by National Natural Science Foundation of China (No. 30028008); National Foundation of Basic Science Research (No.G2000057004 and G1999054202).

摘　　要：目的观察烧伤大鼠血清刺激人脐静脉内皮细胞株ECV-304引起的纤维状肌动蛋白在细胞内分布变化的时间效应以及肌球蛋白轻链激酶在其中的作用。方法用烧伤大鼠血清分别孵育细胞30 min、1 h、2 h、4 h和6 h。或在烧伤血清作用30 min之前或之后使用5 μmol/L的ML-7作用30 min。使用罗丹明标记的鬼笔环肽探针特异性结合并显示细胞内的纤维状肌动蛋白,并在Nikon TE-300荧光显微镜下观察照相。结果正常情况下,纤维状肌动蛋白主要分布在内皮细胞内的外周膜部位。使用烧伤大鼠血清进行30 min^12 h的刺激后,内皮细胞内可观察到明显的应力纤维形成,并且随着刺激时间的延长而增强。可以通过使用肌球蛋白轻链激酶特异性抑制剂ML-7预处理细胞30 min来抑制这种效应。此外,ML-7还能够改善烧伤大鼠血清引起的内皮细胞内肌动蛋白重排。结论本研究表明,大鼠烧伤血清能够通过激活肌球蛋白轻链激酶引起明显的细胞内肌动蛋白排列的改变。在烧伤血清刺激细胞后,抑制肌球蛋白轻链激酶可以改善烧伤大鼠血清引起的内皮细胞骨架重排。Objectives To investigate the time-dependent effects of serum from burned rats on cytoskeletal filamentous actin (F-actin) reorganization by visualizing their distribution in human umbilical vein endothelial cell line ECV-304 and evaluate the role of myosin light-chain kinase (MLCK) in this process. Methods The serum-starved ECV-304 cells were incubated with the serum from burned rats for 30 min, 1, 2, 4, and 6 h, respectively, and 30 min before or after the incubation, the cells were treated with 5 μmol/L ML-7 for 30 min. F-actin was stained with rhodamine-phalloidin and observed under fluorescence microscope. Results Under normal condition, F-actin was distributed mainly in the cortical area of the endothelial cells. After stimulation with the burn serum, stress fiber formation could be clearly seen in the endothelial cells, exhibiting a time-dependent enhancement in a time course ranging from 30 min to 6 h. Such an effect could be significantly inhibited by a 30-min pretreatment of the cells with MLCK-specific inhibitor ML-7. Inhibition of MLCK also reversed actin reorganization in the endothelial cells pretreated with the burn serum. Conclusion Serum from burned rats induces characteristic morphological changes in the endothelial cell actin cytoskeleton mainly due to the MLCK activation, an effect that can be reversed by the inhibition of MLCK.

关 键 词：肌球蛋白轻链激酶 烧伤 血清 内皮细胞骨架 

分 类 号：R644[医药卫生—外科学]