同型半胱氨酸对血管内皮细胞骨架的影响  

Effects of homocysteine on the cytoskeleton of vascular endothelium

作　　者：彭红艳[1] 常青[1] 唐海兰[2] 李自成[3] 

机构地区：[1]暨南大学医学院组胚教研室,广州510632 [2]暨南大学中心实验室 [3]暨南大学第一临床学院心内科

出　　处：《临床心血管病杂志》2008年第4期294-297,共4页Journal of Clinical Cardiology

基　　金：国家自然科学基金资助项目(No:30240076)

摘　　要：目的:研究同型半胱氨酸(Homocysteine,Hcy)对内皮细胞形态及细胞骨架肌动蛋白的影响,并探讨骨架变化与内皮损伤的关系。方法:将体外培养的ECV-304内皮细胞株随机分为正常对照组、低剂量Hcy组、中剂量Hcy组和高剂量Hcy组。倒置相差显微镜观察各组培养24h的内皮细胞的形态变化;并用Rhodamine-Phalloidin标记纤维状肌动蛋白(F-actin),Alexa flour488-DnaseⅠ标记球状肌动蛋白单体(G-actin),激光扫描共聚焦显微镜(LSCM)观察各组内皮细胞骨架肌动蛋白的不同变化。结果:正常对照组内皮细胞基本为梭形,呈典型的鹅卵石样外观;各Hcy组内皮细胞形态则明显不同,随着Hcy浓度的增加,胞体出现收缩,细胞变圆、渐至不规则,且轮廓增强,细胞间隙增大,甚至坏死脱落。LSCM下发现,正常对照组内皮细胞F-actin在周边形成致密周围带,胞质内有部分应力纤维;随着Hcy浓度的递增,外周致密带逐渐出现断裂、甚至消散;胞质内应力纤维变短、增粗,最终消失。而G-actin正常时仅局限在核周,在受到不同浓度Hcy作用后,明显向外周扩散,且荧光强度增大。结论:内皮细胞骨架肌动蛋白的形态变化与重排,可能是Hcy损伤内皮细胞的重要机制之一。Objective：To investigate the effects of homocysteine （Hcy） on morphology and cytoskeleton-actins of endothelial cells,and discuss the relationship between cytoskeleton change and endothelial injury. Method, ECV- 304 （an endothelial cell strain） cells were cultivated with DMEM in vitro, and then divided randomly into normal control group, low-dose Hcy group （0.1 mmol Hcy/L） ,middle -dose Hcy group（0.5 mmol Hcy/L） and high-dose Hcy group（1.0 mmol Hcy/L）. 24 hours later, the morphologic changes of endothelial cells were observed in inverted phase contrast microscope. And then Rhodamine-Phalloidin was used for labelling filamentous actin （F-actin）, Alexa flour 488 -Dnase Ⅰ was used for labelling globular monomeric antin （G-actin）, the morphology and the distribution of F-actin and G-actin were observed with laser scanning confocal microscope （LSCM）. Result： The shape of endothelial cells of control group were fusiform,arranging like the typical cobblestone appearance; The cells of Hcy group undergone distinct morphologic changes： cells were getting contracted and the intercellular spaces increased with the increase of Hcy concentration, even necrosis and defluvium appeared. The F-actins in control group were concentrated and closed to the cell periphery in which form dense periphery band, and there were some stress fibers in the cytoplasm. Dense periphery band gradually showed fragmentation and debris, the stress fibers in the cytoplasm were decurtated and thicken, and ultimately disappeared in the high-dose Hey group. G-actins of control group were limited in periphery distrit of the nuclei, with the increase of Hcy concentration, Gactin gradually dispersed, and fluorescence intensity increased. Conclusion：The morphologic change and rearrangement of cytoskeleton-actins may be one of the important mechanisms of the damage of Hcy on endothelial cells induced by Hcy.

关 键 词：细胞骨架 内皮 血管 同型半胱氨酸 肌动蛋白 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]