S1P/S1PR信号通路对中性粒细胞功能的影响  

Effect of Sphingosine 1-Phosphate/Sphingosine 1-Phosphate Receptor Signal Pathway on Function of Neutrophils

作　　者：王中英[1,2] 谢如锋[2] 杨洁[2] 任亚娜[2] 杨懿铭[2] 范华骅[2] 

机构地区：[1]华东师范大学生命科学学院生物医学系,上海200062 [2]上海市血液中心血液工程研究室,上海200051

出　　处：《中国实验血液学杂志》2012年第4期989-994,共6页Journal of Experimental Hematology

摘　　要：本研究旨在检测1-磷酸鞘氨醇(S1P)对fMLP活化的中性粒细胞的引发作用,检测粒细胞活化后发生呼吸爆发的产物,并探究S1P引发作用的信号通路。流式细胞术检测新鲜分离的中性粒细胞的状态;高铁细胞色素C还原法间接计算超氧阴离子(O2-)释放量;二氢罗丹明123流式细胞术检测粒细胞的呼吸爆发程度;免疫印迹法检测PI3K-Akt信号通路相关蛋白的表达。结果表明,S1P预处理后fMLP活化的粒细胞所释放的O2-量明显增高;S1P引发的fMLP活化组中,罗丹明123的平均荧光强度明显高于fMLP单独处理组;PI3K及Akt蛋白参与了粒细胞呼吸爆发的信号传递。结论:S1P是一种新的粒细胞引发试剂,较高浓度S1P(10-6μmol/L)能够引发fMLP活化的中性粒细胞发生呼吸爆发;该信号通路可能为S1P与粒细胞上的S1P受体作用,活化下游的PI3K,再通过Akt传递信号,最终活化NADPH氧化酶,发生呼吸爆发。The aim of this study was to examine the priming effect of sphingosine 1-phosphate （S1P） on fMLP- activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometly was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine123 mean fluorescence intensity in SiP primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that SIP is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3 K, then activates Akt-transmiting signals through NADPH oxidase, finally results in the respiratory burst.

关 键 词：中性粒细胞 呼吸爆发 1-磷酸鞘氨醇 超氧阴离子 磷酸肌醇-3-激酶 

分 类 号：R331.142[医药卫生—人体生理学] R392.12[医药卫生—基础医学]