原花青素通过对Caspase-3和Bcl-xl、Bax蛋白的调节抑制人脐静脉内皮细胞凋亡  

Inhibition of Procyanidins on the Apoptosis of Human Umbilical Vein Endothelial Cell through the Regulating Caspase-3 and Bcl-xl and Bax

作　　者：丁黎敏[1] 黃小民[1] 张微[1] 陆如凤[1] 

机构地区：[1]浙江中医药大学附属第一医院急诊科,浙江杭州310006

出　　处：《中华中医药学刊》2013年第3期582-584,I0007,I0008,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省医药卫生科学研究基金资助项目(2007A146)

摘　　要：目的:用过氧化氢建立人脐静脉内皮细胞(HUVEC)凋亡模型,加以原花青素(procyanidin)干预,研究原花青素是否对过氧化氢诱导的内皮细胞凋亡有抑制作用并探讨其机制。方法:体外培养人脐静脉内皮细胞,将细胞分为6组,即空白对照组(Control)、氧化损伤组(H2O2)、氧化损伤加入VitC对照组(VC+H2O2)、氧化损伤加入原花青素5、25、50μmol/L浓度组(procyanidin-L+H2O2、procyanidin-M+H2O2、procyanidin-H+H2O2)。将750μmol/LH2O2作用于加入VitC及不同浓度原花青素预培养24 h的内皮细胞,继续培养18 h(半胱氨酸蛋白酶表达测定时间为14 h),以吖啶橙/溴化乙锭(AO/EB)双荧光染色显微镜观察法、AnnexinV/PI流式细胞术检测细胞凋亡,并采用Western blot方法检测Caspase-3、Bcl-xl、Bax蛋白的表达。结果:原花青素呈剂量依赖性抑制H2O2诱导的内皮细胞的凋亡,并显著下调Caspase-3、Bax蛋白表达,明显上调Bcl-xl蛋白表达,并呈剂量依赖性,各指标差异有显著性(P<0.01)。结论:原花青素能抑制过氧化氢诱导的血管内皮细胞的凋亡,其作用可能与下调Casapase-3表达,升高Bcl-xl/Bax比值有关。Objective：The human umbilical vein endothelial cells（HUVEC）apoptosis model was established by hydrogen peroxide and intervented by procyanidins（procyanidin）.The effect of procyanidins on hydrogen peroxide-induced endothelial cell apoptosis and its mechanism were explored.Methods：The human umbilical vein endothelial cells were cultured and divided into six groups,the control group（Control）,oxidative damage group（H2O2）,oxidative combined with VitC control group（VC ＋ H2O2）,oxidative combined with procyanidins at doses of 5 μmol/L,25 μmol/L,50 μmol/L groups（procyanidin-L ＋ H2O2,procyanidin-M ＋ H2O2,procyanidin-H ＋ H2O2）.The 750 μmol/L H2O2 was added to the cells which were cultured with VitC and different concentrations of procyanidins for 24 hours,culturing for 18 hours（cysteine protease expression determination time was 14 hours）,acridine orange/ethidium bromide（AO/EB）dual staining microscope observation,Annexin V/ PI flow cytometry were used to detect cell apoptosis,and Western blot was used for analysis of Caspase-3,Bcl-xl and Bax protein expressions.Results：The procyanidins can dose-dependently inhibit H2O2-induced apoptosis of endothelial cells,and significantly lower the expressions of Caspase-3 and Bax protein,significantly up-regulate Bcl-xl protein expression,in a dose-dependent manner,all indicators showed the difference was significant（P〈0.01）.Conclusion：The procyanidins can inhibit vascular endothelial cell apoptosis induced by hydrogen peroxide,which may be related with the down-regulating casapase-3 expression and increasing Bcl-xl/Bax ratio.

关 键 词：原花青素 过氧化氢 内皮细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学]