铜离子对血管内皮细胞增殖与分化的影响  

EFFECT OF COPPER-ION ON PROLIFERATION AND DIFFERENTIATION OF VASCULAR ENDOTHELIAL CELLS

作　　者：罗堃[1] 范兆心[1] 冯喆[1] 李顺[1] 陈晓禾[1] 解慧琪[1] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室.干细胞与组织工程研究室,成都610041

出　　处：《中国修复重建外科杂志》2009年第7期832-835,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划(863)资助项目(2009AA02Z106);教育部新世纪优秀人才支持计划(NCET-07-575)~~

摘　　要：目的评价Cu2+对人脐静脉血管内皮细胞(human umbilical vein endothelial cell,HUVEC)增殖与分化的影响。方法体外培养并传代HUVEC。将HUVEC以5×103个/孔密度接种于96孔板,根据向孔板中加入溶液浓度的不同将细胞随机分成3组,A组5μmol/L CuSO4,B组25μmol/L CuSO4,C组为空白对照,每组4个复孔,基础培养基为MCDB131,采用MTT法检测细胞增殖并绘制生长曲线。另取HUVEC以2×105个/孔密度接种于6孔板,分组同前,荧光定量RT-PCR检测3组HUVEC的内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)基因和Tie-1基因表达。结果生长曲线示前3d为指数增长期,第4天开始进入平台期。A组细胞增殖能力从第3天开始明显高于B、C组,B组在2d内高于C组,但从第4天开始显著低于C组,比较差异均有统计学意义(P<0.05)。荧光定量RT-PCR检测示作用48h后,A、B、C组eNOS基因表达分别为7.294±1.488、0.149±0.044和1.000±0.253,Tie-1基因的表达分别为1.481±0.137、1.131±0.191和1.000±0.177。A组与B、C组比较,2个基因的表达均有上调(P<0.05);B、C组比较,eNOS基因表达下调(P<0.05),Tie-1基因表达差异无统计学意义(P>0.05)。结论5μmol/L Cu2+能有效促进HUVEC的增殖与分化。Objective To evaluate the effect of copper-ion on the proliferation and differentiation of human umbilical vein endothelial cell （HUVEC）. Methods HUVEC were cultured and passaged in vitro. HUVEC were inoculated into 96-well plate with density of 5 × 10^3/well. All the cells were divided into 3 groups randomly according to different culture mediums： group A （5 μmol/L CuSO4）, group B （25 μmol/L CuSO4）, group C （control group）. Every group had 4 wells, and the basic culture medium was MCDB131. The cell growth curves of 3 groups were drawn by using MTT. HUVEC were inoculated into 6-well plate with density of 2 × 10^5/well. Grouping of the cells was the same as the above. The gene expressions of endothelial nitric oxide synthase （eNOS） and tyrosine kinase with immunoglobulin-like and EGF-like domain 1 （Tie-1） were detected by real-time RT-PCR. Results The growth curves revealed that the exponential growth time was the first 3 days, plateau growth time begun on the 4th day. The proliferation of group A was stronger than that of groups B and C from the 3rd day, within 2 days, the proliferation of group B was stronger than that of group C; however, it decreased and was weaker than group C from the 4th day, all showing statistically significant difference （P 〈 0.05）. The results of real-time RT-PCR revealed that the expressions of eNOS in groups A, B and C were 7.294 ± 1.488, 0.149 ± 0.044 and 1.000 ± 0.253; and the expressions of Tie-1 in groups A, B and C were 1.481 ± 0.137, 1.131 ± 0.191 and 1.000 ± 0.177. Group A compared with groups B and C, both of 2 genes were up-regulated （P 〈 0.05）. Group B compared with group C, eNOS was down-regulated （P 〈 0.05） and the difference of Tie-1 expression was not statistically significant （P 〉 0.05）. Conclusion 5 μmol/L copper-ion can promote the proliferation and differentiation of HUVEC effectively.

关 键 词：CU^2+ 人脐静脉血管内皮细胞 细胞增殖 基因表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]