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|本期目录/Table of Contents|

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

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2010年第1期
页码:
1-5
栏目:
基础研究
出版日期:
2010-01-04

文章信息/Info

Title:
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; 梁春; 任雨笙; 姜其均; 吴建祥; 吴宗贵
第二军医大学附属长征医院心内科，上海 200003
Author(s):
JIANG Hai-long;  LIANG Chun;  REN Yu-sheng;  JIANG Qi-jun;  WU Jian-xiang;  WU Zong-gui
Department of Cardiology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
关键词:
软脂酸; 通心络; 内皮祖细胞; caspase-3
Keywords:
palmitic acid;  tongxinluo;  endothelial progenitor cells;  caspase-3
分类号:
R329.25
DOI:
-
文献标识码:
A
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(endothelial progenitor 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7 d后，贴壁细胞分成7组：对照组以M199培养液培养48 h；PA各浓度组(共3组)分别用含200，400，800 μmol/L PA的M199培养液孵育48 h；通心络干预组(共3组)分别先用含100，200，400 μmol/L TXL的M199培养液干预2 h后，再加入400 μmol/L PA孵育48 h。采用流式细胞仪检测细胞凋亡率；提取细胞蛋白，采用Western blot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400 μmol/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。
Abstract:
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 h］ and three TXL groups ［attached cells were incubated with different concentrations of TXL (100, 200 and 400mol/L), respectively, for 2 h before incubation with 400 μmol/L PA for 48 h］. 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.

参考文献/References

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备注/Memo

备注/Memo:
收稿日期：2008-12-30.基金项目:国家重点基础研究规划项目973基金资助（2005CB523309） 通讯作者:吴宗贵，教授，主要从事动脉粥样硬化研究Email:zgwu@medmail.com.cn 作者简介:江海龙，博士生Email:bluefate121@126.com
更新日期/Last Update: 2010-01-05