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鞘氨醇-1-磷酸/鞘氨醇-1-磷酸受体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].

作者信息

Wang Zhong-Ying, Xie Ru-Feng, Yang Jie, Ren Ya-Na, Yang Yi-Ming, Fan Hua-Hua

机构信息

Department of Biology and Medicine, East China Normal University, Shanghai, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2012 Aug;20(4):989-94.

Abstract

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 cytometry 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 Rhodamine 123 mean fluorescence intensity in S1P 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 S1P 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 PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.

摘要

本研究的目的是检测1-磷酸鞘氨醇(S1P)对fMLP激活的中性粒细胞的启动作用,主要检测中性粒细胞呼吸爆发产物,并研究S1P活性涉及的信号通路。采用流式细胞术评估新分离的中性粒细胞;通过呼吸爆发中细胞色素C还原间接检测超氧阴离子输出;用二氢罗丹明123检测呼吸爆发强度;通过蛋白质免疫印迹法探索中性粒细胞呼吸爆发的信号转导途径。结果显示,用S1P预处理后,fMLP激活的中性粒细胞释放的超氧阴离子水平显著升高;S1P启动的fMLP激活的中性粒细胞组中罗丹明123平均荧光强度显著高于fMLP处理组;PI3K和Akt蛋白参与中性粒细胞呼吸爆发信号通路。结论是,S1P是一种新的启动试剂,可启动fMLP激活的中性粒细胞的呼吸爆发;该信号通路可能是S1P与其受体相互作用,激活PI3K,然后通过NADPH氧化酶激活Akt传递信号,最终导致呼吸爆发。

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