雌激素诱导的合成型血管平滑肌细胞凋亡与p38通路激活及肌细胞增强因子-2的表达  

MOLECULAR MECHANISMS OF ESTROGEN INDUCED APOPTOSIS OF SYNTHETIC VASCULAR SMOOTH MUSCLE CELLS: p38 PATHWAY-DEPENDENT UP-REGULATION OF MYOCYTE ENHANCER FACTOR 2

作　　者：程蓓[1] 王乔[1] 雷岳山[1] 宋健[1] 

机构地区：[1]武汉大学医学院人体解剖学与组织胚胎学系,武汉430071

出　　处：《解剖学报》2007年第6期718-721,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30470906)

摘　　要：目的探讨雌激素诱导合成型血管平滑肌细胞(VSMCs)凋亡的分子机制。方法体外培养第3代的雌性大鼠VSMCs分为3组:17β-雌二醇(E2)组、p38通路阻断(SB+E2)组和对照组。药物处理72h后,ELISA法检测细胞内核小体以检测VSMCs凋亡率,Western blotting检测p38总蛋白、磷酸化p38(p-p38)的变化及肌细胞增强因子-2(MEF2)的表达,免疫组织化学染色确认MEF2的定位和表达。结果ELISA检测显示,E2组VSMCs凋亡率明显高于对照组,而SB+E2组无明显变化。Western blotting显示,E2组p38、p-p38和MEF2均明显增高,而SB+E2组p38表达无明显变化,p-p38和MEF2明显降低。免疫组织化学染色显示,MEF2主要定位于细胞核,各组表达水平与Western blotting结果一致。结论雌激素可能通过激活p38通路上调MEF2的表达,诱导VSMCs凋亡。Objective To explore the underlying molecular mechanisms of estrogen induced apoptosis of synthetic vascular smooth muscle cells （VSMCs）. Methods Cultured VSMCs on passage 3 were divided into three groups： 17β-estradiol （ E2 ） group, p38 inhibitor group （ SB ＋ E2 ） and control group. With treatment for 72 hours, the apoptosis of cells was quantified by measuring the intracellular nucleosomes with ELISA; expressions of total p38, phospho-p38 and myocyte enhancer factor-2 （MEF2） were analyzed by Western blotting, with a contlrmative immunohistochemical staining for MEF2. Results Compared with that of control cells, the rate of apoptosis in VSMCs grown in E2 was significantly increased, with up-regulations of total p38, phospho-p38 and MEF2. These effects of E2 were blocked in SB ＋ E2 group in which the rate of cell apoptosis and expression of p38 had no obvious change compared with that of control group, while the expression of p-p38 and MEF2 was decreased. Immunohistochemical staining showed that MEF2 was mainly located in cellular nucleus, and its expression was increased in E2 group but decreased in SB ＋ E2 group. Conclusion Estrogen may induce apoptosis of synthetic VSMCs through p38 pathwaydependent up-regulation of MEF2.

关 键 词：雌激素 平滑肌细胞 P38通路 肌细胞增强因子-2 免疫组织化学 免疫印迹法 大鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]