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

作　　者：薛晓婕[1] 刘佳佳[2] 

机构地区：[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

出　　处：《放射免疫学杂志》2013年第1期63-65,共3页Journal of Radioimmanology

基　　金：国家自然科学基金资助项目(30371305)

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(Polymorphonuelearneutrophils,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-αmRNA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-αmRNA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。Objective To explorc the role of Bax-α in non-apoptotic PCD of neutrophils induced by ONO-AE-248. Methods Neutrophils from peripheral venous blood of healthy volunteers with fieoll method were cultured in vitro with ONO-AE-248 （5 × 10-5 mol/L） for 12h. The activation of Bax-α of neutrophils stimulated with medium alone, LPS or TNF-α and ONO-AE-248 was observed. The viability of neutrophils was detected by flow-cytometric analysis of double staining with Annexin V-FITC and propidium iodide （PI）. The expression level of Bax-α was detected in ONO-AE-248-stimulated 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. Gonclusion Bax-α 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 

分 类 号：R329.12[医药卫生—人体解剖和组织胚胎学]