ONO-AE-248对诱发中性粒细胞非凋亡性程序化死亡中Bax-α表达的研究
Study on the Expression of Bax-α of Non-apoptotic PCD of Neutrophils Induced by ONO-AE-248
薛晓婕 1刘佳佳2
作者信息
- 1. 湖北省黄石市中心医院检验科,435000
- 2. 泸州医学院分子生物学实验室,646000
- 折叠
摘要
目的:为探讨接受ONO-AE-248 刺激后重要的表达蛋白-Bax-α在中性粒细胞(Polymorphonuelear neutrophils,PMN)非凋亡程序化细胞死亡中的表达及意义,以进一步确定这种新型细胞死亡模式提供线索.方法:采用Ficoll法新鲜分离健康志愿者外周静脉血PMN与 ONO-AE-248 共同培养12 h后,流式观察ONO-AE-248对细胞活性的影响并提取总RNA,并行反转录-PCR检测Bax-α mRNA的表达;Western- blot检测Bax-α蛋白的表达.结果:ONO-AE-248 刺激 12 h时PMN Bax-α mRNA的表达与空白组比较无明显变化(P>0.05);蛋白印迹法显示,Bax-α蛋白的表达无差异;流式结果显示,ONO-AE-248刺激的中性粒细胞发生快速死亡.结论:Bax-α在ONO-AE-248诱导的非凋亡性程序化死亡中可能发挥着部分相同或相关的作用.
Abstract
Objective To explore the role of Bax-a in non-apoptotic PCD of neutrophils induced by ONO-AE-248. Methods Neutrophils from peripheral venous blood of healthy volunteers with ficoll method were cultured in vitro with ONO-AE-248 (5 × 10-5 mol/L) for 12h. The activation of Bax-a of neutrophils stimulated with medium alone, LPS or TNF-α and ONO-AE-248 was observed. The viability of neutrophils was detected by flow-cytoraebic analysis of double staining with Annexin V-FTTC and propidium iodide (PI). The expression level of Bax-a was detected in ONO-AE-248-stunulated PMN by RT-PCR and Western blot technic. Results PMN cound express the mRNA and protein of Bax-α, the expression of Bax-α expressed no significant changes between ONO-AE-248 group with medium alone group(P＞0.05). Flow-cytometric analysis showed PMN were induced apoptosis rapidly by ONO-AE-248. Conclusion Bax-a played the same or like-pathway effect between the two kinds of cell death.
关键词
中性粒细胞/ONO-AE-248/非凋亡性程序化细胞死亡/Bax-α/凋亡Key words
neutrophils/ONO-AE-248/non-apoptotic programmed cell death/Bax-a/apoptosis引用本文复制引用
出版年
2013
NETL
NSTL
维普
万方数据