同型半胱氨酸诱导细胞周期G1/S阻滞抑制人脐静脉内皮细胞的增殖  

Homocysteine Inhibits Human Umbilical Vein Endothelial Cells Proliferation through Cell Cycle G1 / S Arrest

作　　者：李倩[1] 闫婷婷[1] 张泉[1] 李林[1] 吴伟康[1] 谭红梅[1] 

机构地区：[1]中山大学中山医学院病理生理教研室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2010年第6期767-771,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学青年基金(30600250);人事部留学回国人员基金(2006;164#)

摘　　要：【目的】探讨同型半胱氨酸(Hcy)对人脐静脉内皮细胞(HUVEC)增殖和细胞周期的影响及其可能的机制。【方法】HUVEC细胞采用含200mL/L胎牛血清、0.06g/L内皮细胞生长添加剂(ECGS)、0.05g/L肝素的M199培养。分为对照组(Control)、50μmol/L腺苷(Ade)组(A50)、50μmol/LHcy组(H50)以及Ade50μmol/L+Hcy50μmol/L组(A50H50)。分别通过细胞计数和MTT法观察Hcy对HUVEC增殖的影响,流式细胞仪检测Hcy对HUVEC细胞周期的影响,WesternBlot检测Hcy对细胞外信号调节蛋白激酶(ERK)及磷酸化状态的影响。【结果】A50H50组的细胞数与细胞增殖活性显著低于对照组、A50组和H50组(P<0.05)。A50H50组G1期细胞百分比显著高于对照组,S期细胞百分比则显著低于对照组(P<0.05),出现明显的G1/S期阻滞。A50H50组ERK磷酸化水平明显低于对照组。【结论】Hcy(50μmol/L)和Ade(50μmol/L)联合使用可引起HUVEC细胞周期的G1/S阻滞,抑制HUVEC细胞的增殖,其机制可能与ERK的磷酸化水平降低有关。【Objective】 To investigate the effect of moderate concentration of homocysteine （Hcy） on human umbilical vein endothelial cells （HUVEC） proliferation,cell cycle,and possible signaling mechanisms. 【Method】 HUVEC were cultured in M199 medium supplemented with 20% fetal bovine serum,0.06 g / L endothelial cell growth supplement and 0.05 g / L heparin. HUVEC were divided into control,adenosine （50 μmol / L,A50）,Hcy （50 μmol / L,H50） and Ade plus Hcy （A50H50） groups. Cell proliferation was assessed by cell counting and MTT method. Cell cycle distribution was assessed by flow cytometry. The phosphorylation status of extracellular signal-regulated protein kinase （ERK） was detected by Western blot analysis. 【Results】 Cell number and viability were significantly decreased in A50H50 group compared with control,A50 and H50 groups （P 0.05）. The percentage of G1 phase cells of A50H50 group was significantly increased,while the percentage of S phase cells was significantly decreased compared with control group （P 0.05）,which represented an obvious G1 / S arrest in A50H50 group. The phosphorylated ERK of A50H50 group was significantly decreased compared with the other 3 groups. 【Conclusion】 Hcy combined with Ade induced G1 / S arrest leading to impaired HUVEC proliferation,which was associated with decreased phosphorylation of ERK.

关 键 词：同型半胱氨酸 人脐静脉内皮细胞 细胞周期 细胞外信号调节蛋白激酶 

分 类 号：R318.11[医药卫生—生物医学工程]