摘要
目的 研究二烯丙基二硫化物 (DADS)诱导CNE2细胞凋亡及 p38MAPK信号转导通路对此过程的作用。 方法 DADS处理CNE2细胞 2 4h后 ,荧光显微镜下观察形态学变化及凋亡细胞计数 ,MTT法测定细胞活性 ,流式细胞仪检测凋亡细胞 ,蛋白质印迹法检测磷酸化p38MAPK表达。结果 在培养的CNE2细胞中 ,DADS(50～ 1 50 μmol·L- 1 )作用 2 4h后 ,DADS诱导CNE2细胞产生典型的凋亡细胞形态学变化 (核浓染 ,核碎裂 ) ,流式细胞仪结果显示 ,随着DADS给药剂量增加 ,细胞周期中各期细胞所占百分率的变化无规律 ,细胞凋亡呈剂量依赖性 ,DADS(50～ 1 50 μmol·L- 1 )浓度依赖性刺激磷酸化p38MAPK的表达 ,p38MAPK抑制剂SB2 0 3580明显增强DADS致凋亡作用。结论 DADS诱导CNE2细胞凋亡时激活磷酸化 p38MAPK表达 。
AIM To investigate the effects of p38 MAPK signaling on diallyl disulfide ( DADS ) induced apoptosis in CNE2 cells. METHODS Morphological changes and quantification of apoptotic cells were determined under fluorescence microscope after a 24 h treatment of CNE2 by DADS. Cell viability was determined with MTT method. Apoptosis detection was taken to the cells on flowcytometry. The expression of phospho p38 MAPK was measured by Western blotting. RESULTS After treatment with DADS at 50～150 μmol·L -1 for 24 h,DADS elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation). The amount of apoptosis cells increased in a concentration dependent manner but cell cycle arrest did not at 50～150 μmol·L -1 ,incubation of CNE2 with DADS for 24 h also induced phospho p38 MAPK expression in a concentration dependent manner. Interestingly, DADS induced apoptosis was markedly increased by preincubation with SB203580, a specific p38 MAPK inhibitor,and increased the expression of phospho p42/ p44 CCDPK in a concentration dependent manner. CONCLUSION DADS activates p38 MAPK,and phospho p38 MAPK signaling appears to play protective roles in DADS induced apoptosis in CNE2.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2003年第4期418-423,共6页
Chinese Pharmacological Bulletin
基金
中南大学归国留学人员专项基金资助课题No181176 0 59
