摘要
目的:研究谷胱甘肽(GSH)对人离体嗜中性白细胞超氧阴离子产生和清除的影响及机制。方法:用细胞色素C还原法测定3种刺激剂:N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(fMLP)、佛波豆蔻醚乙酸盐(PMA)和花生四烯酸(AA)介导的嗜中性白细胞超氧阴离子释放量。用Western blot法检测嗜中性白细胞蛋白质因子p47phox磷酸化。结果:GSH呈浓度依赖性促进fMLP介导的嗜中性白细胞释放超氧阴离子,不影响PMA和AA介导的嗜中性白细胞释放超氧阴离子。酪氨酸激酶抑制剂5,7,45-三羟基异黄酮明显抑制fMLP介导的氧自由基产生,而蛋白激酶C抑制剂十字孢碱仅有轻度抑制作用。在非初始化的嗜中性白细胞GSH可清除fMLP、PMA和AA诱导产生的超氧阴离子。GSH刺激fMLP介导的嗜中性白细胞质因子p47phox磷酸化,与其对超氧阴离子释放的影响一致。结论:GSH对刺激剂介导的超氧阴离子的产生依赖于膜受体的激活和还原型辅酶Ⅱ(NADPH)氧化酶p47phox磷酸化。
Objective：To study the effect of glutathione （GSH） on stimulus-induced superoxide anion generation and scavenging in human neutmphils in vitro and the mechanism. Methods： The superoxide anion generation in human neutrophils induced by N-fonnyl-methionylleucyl-phenylalanine （tMLP）, phorbol 12-myristate 13-acetate （PMA）, arachidonic acid （AA） was determined by cytechrome C reduction method. The phosphorylation of p47^phox in neutrophils was detected by Western blot. Results：GSH increased the superoxide anion generation induced by fMLP in a concentration-dependent manner, but had no obvious effect on the superoxide anion generation induced by PMA and AA. fMLP-induced superoxide anion generation was strongly inhibited by genistein,the inhibitor of tymsine kinase,and slightly inhibited by staurosporine,the inhibitor of pwtein kinase C. GSH can scavenge fMLP-,PMA- and AA-induced superoxide anion generation in unprimed neutmphils. GSH increased fMLP-induced p27^phox phosphorylation in a concentration-dependent manner. Conclusion： The effect of GSH on stimulus-induced superoxide anion generation is dependent on the activation of membrane receptor-tymsine kinase pathway and the phospho- rylation of p47^phox of NADPH oxidase.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2006年第6期634-636,共3页
Journal of China Medical University
基金
教育部回国人员启动基金资助项目(教外司留[2004]527号)
