通心络下调caspase-3蛋白表达及活性抑制软脂酸诱导的人外周血内皮祖细胞凋亡  

Tongxinluo inhibits palmitic acid-induced apoptosis of human endothelial progenitor cells by down-regulating expression and activity of caspase-3 protein

作　　者：江海龙[1] 梁春[1] 任雨笙[1] 姜其均[1] 吴建祥[1] 吴宗贵[1] 

机构地区：[1]第二军医大学附属长征医院心内科,上海200003

出　　处：《心脏杂志》2010年第1期1-5,共5页Chinese Heart Journal

基　　金：国家重点基础研究规划项目973基金资助(2005CB523309)

摘　　要：目的:探讨通心络(tongxinluo,TXL)对软脂酸(palmitic acid,PA)诱导的人外周血内皮祖细胞(endothelialprogenitor cell,EPCs)凋亡及caspase-3蛋白表达和活性的影响。方法:密度梯度离心法获取人外周血单个核细胞,培养7d后,贴壁细胞分成7组:对照组以M199培养液培养48h;PA各浓度组(共3组)分别用含200,400,800μmol/L PA的M199培养液孵育48h;通心络干预组(共3组)分别先用含100,200,400μmol/L TXL的M199培养液干预2h后,再加入400μmol/L PA孵育48h。采用流式细胞仪检测细胞凋亡率;提取细胞蛋白,采用Westernblot技术检测caspase-3蛋白表达水平;采用分光光度法检测caspase-3蛋白活性水平。结果:PA呈浓度依赖性诱导EPCs凋亡;加入TXL干预,EPCs细胞凋亡率明显降低;Western blot和分光光度法检测显示,PA组EPCs的caspase-3蛋白表达及活性明显高于对照组,200和400μmol/L TXL干预组明显低于PA组(P<0.05)。结论:PA呈浓度依赖性诱导EPCs凋亡,TXL能部分抑制PA的上述作用,其机制与下调caspase-3蛋白表达及活性有关。AIM： To explore the effect of tongxinluo （TXL） on apoptosis, expression and activity of caspase-3 in palmitic acid （PA）-induced peripheral blood-derived human endothelial progenitor cells in vitro. METHODS： Total mononuclear cells were isolated from peripheral blood in vitro by Ficoll density gradient centrifugation and plated on fibronectin-coated culture dishes. After 7 days, the attached cells were divided into seven groups： one control group （normal cells） , three PA groups [ attached cells were incubated with different concentrations of PA （200,400 and 800 mol/L） for 48 hi and three TXL groups [ attached cells were incubated with different concentrations of TXL （ 100,200 and 400 mol/L）, respectively, for 2 h before incubation with 400 μmol/L PA for 48 hi. Flow cytometry was used to detect apoptosis. Expression and activity of caspase-3 protein were detected by Western blot and spectrophotometry, respectively. RESULTS： After exposure to PA, the apoptosis rate in treatment groups was dose-dependently higher than in the control group （P 〈 0. 05）. TXL was added at different concentrations before PA incubation and cells in the TXL groups （200, 400 mol/L） showed lower apoptosis rate than in PA groups （P 〈0.05）. Expression of caspse-3 protein and activity of EPCs in TXL groups was lower than in PA groups （P 〈0. 05）. CONCLUSION： PA promotes cell apoptosis by up-regulating expression and activity of caspase-3. TXL partly inhibits the effects of PA.

关 键 词：软脂酸 通心络 内皮祖细胞 easpase-3 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]