同型半胱氨酸对血管内皮细胞骨架与内皮细胞功能的影响  

Effects of homocysteine on the cytoskeleton-actins and function of vascular endothelial cells

作　　者：彭红艳[1] 常青[2] 唐海兰[3] 李自成[4] 

机构地区：[1]武汉科技大学附属天佑医院超声影像科,武汉430064 [2]暨南大学医学院组胚教研室 [3]暨南大学实验技术中心 [4]暨南大学第一临床学院心内科

出　　处：《临床心血管病杂志》2010年第11期859-863,共5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:通过动态观察同型半胱氨酸(Homocysteine,Hcy)作用后的血管内皮细胞功能的变化,以及对内皮细胞骨架肌动蛋白G-actin和F-actin的定量分析,探讨骨架肌动蛋白在Hcy损伤内皮中的作用及机制。方法:将体外培养的人脐静脉内皮细胞株(ECV-304)分为正常对照组、低剂量Hcy组(0.1mmol/L)、中剂量Hcy组(0.5mmol/L)和高剂量Hcy组(1.0mmol/L),观察各组细胞培养24h时其细胞骨架和相关功能的变化;中剂量Hcy组及高剂量Hcy组还观察了6h、12h时的相应变化。分别采用硝酸还原酶法、硫代巴比妥酸法检测各组条件培养基中一氧化氮(NO)、脂质过氧化代谢产物丙二醛(MDA)的含量。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)检测内皮细胞骨架G-actin、F-actin含量,并计算G/F比值。结果:随着Hcy作用时间的延长或剂量的增大,NO含量总体呈下降趋势,而MDA含量增高;G-actin、F-actin总体含量及G/F比值上升,并呈一定的时间、剂量相关性。结论:Hcy可通过改变骨架蛋白G-actin和F-actin含量,以及G/F比值引致内皮细胞功能的损伤,这也可能是其诱导动脉粥样硬化早期变化的重要机制之一。Objective：On the basis of dynamic observations of vascular endothelial cell＇s function,as well as quantitative analysis of cytoskeleton-actins of ECs（Endothelial cells）,to study the potential mechanisms of ECs damage induced by Hcy.Method：Human umbilical vein endothelial cells（ECV-304）cultivated in DMEM were di-vided into control group,low-dose Hcy group（0.1mmol/L）,medium-dose Hcy group（0.5mmol/L）and high-dose Hcy group（1.0mmol/L）.After 24 hincubation with Hcy,the changes of cytoskeleton-actins and related function of ECs were observed in each group,the corresponding changes were observed at 6h,12 hin the medium-dose group and high-dose Hcy group.The concentration of nitric oxide（NO）and malondial dehyde（MDA）in the conditioned medium were measured by Nitrate reductase enzymes and Thibabituric acid（TBA）respectively. The contents of filamentous actin（F-actin）and globular monomeric actin（G-actin）stained with Rhodamine-Phalloidin and Alexa Flour 488-Dnase I respectively were measured with laser scanning confocal microscope（LSCM）,and G/F ratio was counted.Result：The overall trends of NO production reduced,but MDA contents,the overall G-actin content,F-actin content,G/F ratio increased with the prologation of times and increasing of Hcy contents,which suggested a time and content-related changes.Conclusion：Hcy injured function of endothelial cells by changing the contents of G-actin and F-actin,and G/F ratio.It may be one of important mechanisms in inducing early atherosclerosis formation.

关 键 词：细胞骨架 内皮 血管 同型半胱氨酸 肌动蛋白 定量分析 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]