咖啡酸抗H_2O_2诱导的人脐静脉内皮细胞凋亡及机制探讨  

Effects of caffeic acid on apoptosis of HUVEC-12 induced by H_2O_2 and the mechanism involved

作　　者：熊丹[1] 严奉祥[1] 欧和生[1] 

机构地区：[1]南华大学药物药理研究所

出　　处：《中国临床药理学与治疗学》2007年第12期1367-1371,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：湖南省卫生厅基金(204062);湖南省教育厅重点基金(03A040);国家教育部重点基金(204095);国家"十五"攻关项目(02EFN214300457)

摘　　要：目的:用H_2O_2诱导人脐静脉内皮细胞株凋亡建立氧化应激模型,观察咖啡酸对H_2O_2诱导的人脐静脉内皮细胞株(HUVEC-12)凋亡的保护作用并初步探讨其机制。方法:分别用不同浓度的咖啡酸和H_2O_2处理HUVEC-12,通过四甲基偶氮唑盐还原法(MTT法)检测细胞的活力,流式细胞术测定内皮细胞凋亡率,用吖啶橙溴化乙啶荧光染色(AO-EB染色)观察细胞凋亡形态,Western blotting检测NF-κB、P53和Bcl-2的表达。结果:用0.5 mmol/L H_2O_2和不同浓度咖啡酸共同孵育24 h,MTT提示咖啡酸能明显减轻H_2O_2对HUVEC-12的损伤,提高细胞的存活率,流式细胞检测术提示咖啡酸能使H_2O_2诱导的HUVEC-12凋亡率由21.9%±2.1%降至3.8%±1.1%。AO-EB染色提示咖啡酸能降低H_2O_2诱导的细胞凋亡,使凋亡细胞减少;同时NF-κB和P53表达上调,Bcl-2表达下调。结论:不同浓度的咖啡酸对H_2O_2引起的HUVEC-12损伤有保护作用,并呈现一定的浓度依赖性和时效性,其机制可能与调节NF-κB、P53和Bcl-2蛋白的表达有关。AIM： To investigate the effects of caffeic acid on apoptosis of HUVEC-12 induced by H2O2, and to explore the related pharmacological mechanism. METHODS： The ceils were incubated with H2O2（0.5 mmol/L） and caffeic acid （1, 10, 100 μmol/L）. The viability of HUVEC-12 was measured by MTF assay at different concentrations or times. Flow cytometry analysis was used to determine the rate of cell apoptosis. AO/EB dyeing was used to detect the morphological changes of cells. Westem blotting was used to detect the NF-κB, P53 or Bcl-2 protein expression. RESULTS： HUVEC-12 was pretreated withl, 10, 100 μmol/L caffeic acid for 30 minutes followed by 0.5 mmol/L H2O2, the rate of cell survival reduced in dose-dependent manner. Results of flow cytomery demonstrated that the apoptotic rate of cells decreased from 21.9% ± 2.1% to 3.8% ± 1.1%. Spontaneous apoptosis was significantly increased in the cells incubated with caffeic acid, meanwhile, the NF-κB protein was up-regulated, Bcl-2 protein was down-regulated, and P53 protein was up-regulated. CONCLUSION： Caffeic acid could resist the H2O2 -induced cell apoptosis, with the possible molecular mechanism of regulating the NF-κB, P53 and BCL-2 expression.

关 键 词：咖啡酸 人脐静脉内皮细胞 凋亡 NF-ΚB 

分 类 号：R965.2[医药卫生—药理学]