摘要
目的 观察晚期糖基化终产物 (AGE)对人脐静脉内皮细胞 (HUVEC)的增殖活力和细胞内纤维状肌动蛋白 (F actin)形态学的影响 ,探讨AGE导致毛细血管通透性升高的病理生理学基础。方法 采用四唑盐 (MTT)比色法 ,分别检测并绘制正常组和AGE处理组的HUVEC活力曲线 ;并分别于培龄第 1～ 12天进行F actin的荧光染色。结果 AGE组HUVEC的活力曲线与正常组相比显著下移。荧光染色发现 :正常组HUVEC的F actin纤维主要富集于毗邻细胞膜的周边 ,形成典型的鹅卵石样形态 ,细胞质中未见密集的F actin纤维 ;AGE组细胞内的F actin纤维形态和分布发生明显的改变 ,出现应力纤维、丝状伪足和板状伪足三种异常形态 ,至刺激后期 ,大部分细胞周边的F actin纤维消失 ,胞质中出现密集的应力纤维。结论 AGE对内皮细胞的活力有明显的抑制作用。同时 ,AGE促使内皮细胞F actin从位于细胞膜周边的状态转为弥散于整个细胞质的状态 ,内皮细胞收缩 ,胞间裂隙形成 ,可能引起毛细血管通透性升高。
Objective To investigate the pathophysiological facts underlying the increase in capillary permeability by observing the impact of advanced glycosylation end products (AGE) on proliferation activity of human umbilical vein endothelial cell (HUVEC) and on filamentous actin (F-actin) in it morphologically. Methods Cell viability curves of normal and AGE-albumin treated HUVEC were drawn using MTT cell proliferation assay (MTT-CPA). F-actin was examined with fluorescence staining of rhodamine-phalloidin every day from the 1 st day to the 12 th day of culture. Results The viability curve of the AGE-treated group declined significantly as compared with that of the normal group. Observation by fluorescence microscopy showed that the fibers of F-actin in the cells of the normal group concentrated along the cell membrane and no dense fibers were observed in the cytoplasm. Three forms of abnormal F-actin fibers, i.e. stress fibers, filopodia, and lamellipodia, appeared in AGE-treated HUVEC , while most of the peripheral fibers disappeared and dense stress fibers were observed in the cytoplasm during the late stage. Conclusion AGE inhibited significantly the viability of the endothelium and induced the reorganization of actin in it, i.e. the F-actin transformed from concentrated distribution along the membrane to a diffused one in the cytoplasm, which might mediate the cell shrinkage and the subsequent formation of intercellular gaps, followed by increase in capillary permeability.
出处
《中华老年多器官疾病杂志》
2004年第1期41-44,共4页
Chinese Journal of Multiple Organ Diseases in the Elderly
基金
国家重点基础研究规划项目 (973
G2 0 0 0 0 5 70 0 4)
广东省团队计划项目 (10 717)资助
