Wang Zhongying, Fan Huahua, Xie Rufeng, Yang Jie, Ren Yana, Yang Yiming, Li Wei
Blood Engineering Laboratory, Shanghai Blood Center, Shanghai, China.
Acta Haematol. 2015;134(1):49-56. doi: 10.1159/000369291. Epub 2015 Apr 11.
BACKGROUND/AIMS: This study investigated the priming effect of sphingosine 1-phosphate (S1P) on formyl-Met-Leu-Phe-OH (fMLP)-activated neutrophils, by specific analysis of the neutrophil respiratory burst and the signaling pathway involved in S1P activity.
The neutrophil respiratory burst was indirectly detected by the cytochrome c reduction method and the dihydrorhodamine 123 staining method. The signal transduction pathways of neutrophil respiratory burst primed by S1P were detected by Western blotting.
Our results showed that the S1P receptors (S1PRs) 1, 4 and 5 were the predominantly expressed neutrophil S1PRs at the cDNA level. After pretreatment with S1P, the fMLP-activated neutrophils released increased levels of superoxide anions. PI3K and Akt proteins were involved in the signaling pathway of the neutrophil respiratory burst primed by S1P.
The results indicate that S1P is a new priming reagent for neutrophils and primes the respiratory burst of fMLP-activated neutrophils. S1P interacts with its receptors on neutrophils, resulting in NADPH oxidase activation by the PI3K-Akt cell signaling pathway and induction of the neutrophil respiratory burst.
背景/目的：本研究通过对中性粒细胞呼吸爆发及鞘氨醇-1-磷酸（S1P）活性相关信号通路的特异性分析，研究S1P对甲酰甲硫氨酸-亮氨酸-苯丙氨酸-OH（fMLP）激活的中性粒细胞的预激效应。
采用细胞色素c还原法和二氢罗丹明123染色法间接检测中性粒细胞呼吸爆发。通过蛋白质免疫印迹法检测S1P预激的中性粒细胞呼吸爆发的信号转导通路。
我们的结果显示，在cDNA水平上，S1P受体（S1PRs）1、4和5是中性粒细胞中主要表达的S1PRs。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增加。PI3K和Akt蛋白参与了S1P预激的中性粒细胞呼吸爆发的信号通路。
结果表明，S1P是一种新的中性粒细胞预激试剂，可预激fMLP激活的中性粒细胞的呼吸爆发。S1P与其在中性粒细胞上的受体相互作用，通过PI3K-Akt细胞信号通路激活NADPH氧化酶并诱导中性粒细胞呼吸爆发。
