没食子儿茶素没食子酸酯抑制牛主动脉内皮细胞增殖和对细胞周期分布的影响  

Effect of epigallocatechin 3 gallate on cell proliferation and cell cycle of bovine aortic endothelial cells

作　　者：王桂平[1,2] 叶云[2] 潘学宾[1] 刘新艳[1] 

机构地区：[1]广州医学院护理学院药理学教研室,广东省广州市510182 [2]南方医科大学基因研究所,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2009年第2期309-312,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:血管内皮细胞在肿瘤血管生成中起关键作用,没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)可抑制内皮细胞生长和增殖,但其作用机制仍不清楚。目的:观察没食子儿茶素没食子酸酯对牛主动脉内皮细胞增殖和细胞周期分布的影响。设计、时间及地点:以细胞为观察对象的随机分组实验,于2006-08/2008-05在广州医学院完成。材料:出生1d、未哺乳的新生牛,取主动脉内皮细胞进行原代培养。方法:将指数生长期密度为5×107L-1的牛主动脉内皮细胞接种于96孔培养板,实验分为2组:实验组和对照组分别加入以培养基RPMI1640配制的不同浓度的EGCG180μL(终浓度为5,10,20,40,80g/L)及等体积不含EGCG的培养基,全自动酶标仪上测定不同时间3,6,12,24,36h各孔吸光度值,并计算细胞生长抑制率。选择适当的药物浓度和作用时间点进行EGCG抑制作用的半数抑制浓度(IC50)实验。收集对数生长期的牛主动脉内皮细胞,实验组加入不同浓度(10,20,30,40g/L)的EGCG,对照组加入不含药物的培养基,Multicycle软件分析G1/G0期、S期和G2/M期细胞所占比例。主要观察指标:①不同剂量EGCG在不同时间点对牛主动脉内皮细胞生长、增殖的影响。②运用直线回归分析EGCG对牛主动脉内皮细胞抑制的IC50。③流式细胞仪检测EGCG对内皮细胞周期分布的影响。结果:①EGCG呈剂量及时间依赖性抑制内皮细胞的增殖:EGCG作用12h后,细胞抑制率达高峰,此后其抑制率逐渐下降(P<0.05);从10g/L起,EGCG对内皮细胞增殖的抑制效应逐渐增加,40g/L即达高峰(P<0.05);40g/L与80g/L剂量组相比,抑制率无明显差异(P>0.05)。②在3,6,12,24h和36h等时间点EGCG的IC50分别为23.14,18.79,13.23,14.19,17.45g/L,IC50在12h达高峰,时间继续延长,作用并不增加。③各组G2/M期细胞所占比率无明显差异(P>0.05)。S期细胞比率由(58.02±1.21)%下降为(27.62±3.12)%,其中30g/L和40g/L剂量组与对照�BACKGROUND： Vascular endothelial cells play a key role in the process of tumor angiogenesis. Epigallocatechin 3 gallate （EGCG） can inhibit the proliferation of endothelial cells, but the mechanism of EGCG remains poorly understood. OBJECTIVE： To study the effect of EGCG on cell proliferation and cell cycle in bovine arteria endothelial cells （BAECs）. DESIGN, TIME AND SETTING： The randomized experiment based on cells was performed at the Medical College of Guangzhou from August 2006 to May 2008. MATERIALS： Primary aorta endothelial cells from 1 day born cow were primary cultured. METHODS： Exponential growth BAECs （5×10^/L） were cultured in 96-cell plate. The experiment was divided into the experimental and control groups, which were added different concentration of 180 p L EGCG （end concentration： 5, 10, 20, 40, 80 g/L） in RPMI1640 and equal volume RPMI1640, respectively. Optical density was measured by the automatic enzyme-labeled appearance at 3, 6, 12, 24, 36 hours after the treatment of EGCG, and cell growth inhibiting rate and median inhibitory concentration （IC50） were calculated. Cells in logarithmic growth phase were gathered and added EGCG with concentration of 10, 20, 30, 40 g/Lin the experiment group. The cell ratio of G1/G0 S and G2/were analyzed by the Multicycle soft. MAIN OUTCOME MEASURES： Effects of EGCG with different concentrations on cell growth and proliferation of BAECs at various time points were observed. IC50of EGCG was detected by linear regression analysis. The effect of EGCG on cell cycle distribution was measured by flow cytometry. RESULTS： （1）G could inhibit BAECs proliferation in a time and dose-dependent manner. The peak of anti-proliferation action of EGCG occurred at 12 hours, and then gradually decreased （P 〈 0.05）. The rate of inhibition increased gradually at 10 g/L, reached a peak at 40 g/L （P〈 0.05）, there was no significant difference between inhabitation rate of 40 mg group and that of 80 mg group （P〉 0.05）. （2）I

关 键 词：牛主动脉内皮细胞 细胞周期 细胞增殖 EGCG 

分 类 号：R349.53[医药卫生—基础医学]