Sphingosine 1-phosphate in amniotic fluid modulates cyclooxygenase-2 expression in human amnion-derived WISH cells
Sphingosine 1-phosphate in amniotic fluid modulates cyclooxygenase-2 expression in human amnion-derived WISH cells
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DOI：
10.1074/jbc.m300625200
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发表时间：
2003-08-22
影响因子：
4.8
通讯作者：
Bae, YS
中科院分区：
文献类型：
--
作者：
Kim, JI;Jo, EJ;Bae, YS
花生四烯酸的代谢，特别是花生四烯酸（PGs）的产生，已被提出在分娩的调节中发挥关键作用。此外，已经报道了几种细胞外蛋白调节羊膜细胞中PG的合成。在这项研究中，我们发现溶解在羊水中的脂质成分调节WISH人羊膜细胞中PG的合成，并确定其中一种成分为1-磷酸鞘氨醇（S1 P）。WISH细胞表达几种S1 P受体，包括S1 P（1）、S1 P（2）和S1 P（3）。当用S1 P刺激WISH细胞时，PGE（2）合成以浓度依赖性方式增加，在约100 nM时显示最大活性。S1 P处理还引起环氧化酶2（考克斯-2）mRNA和蛋白的上调，这在刺激3-12 h内明显。在S1 P诱导WISH细胞活化的细胞内信号通路方面，我们发现S1 P刺激两种MAPK，ERK和p38激酶。我们研究了这两种MAPK在S1 P诱导的考克斯-2表达中的作用。PD-98059可完全阻断S1 P诱导的考克斯-2表达，但SB-203580不阻断，表明ERK在此过程中起关键作用。转染S1 P（1）或S1 P（3）反义寡核苷酸可抑制S1 P诱导的WISH细胞考克斯-2表达和PGE（2）的产生，表明S1 P（1）和S1 P（3）参与了这一过程。本研究证实了羊水中S1 P的生理作用及其对WISH细胞中考克斯-2表达和PGs合成的调节作用。
The metabolism of arachidonic acid, in particular the generation of prostaglandins (PGs), has been proposed to play a key role in the regulation of labor. Moreover, several extracellular proteins have been reported to modulate PG synthesis in amnion cells. In this study, we found that lipid components dissolved in the amniotic fluid modulate PG synthesis in WISH human amnion cells and identified one of these components as a sphingosine 1-phosphate (S1P). WISH cells express several S1P receptors including S1P(1), S1P(2), and S1P(3). When WISH cells were stimulated with S1P, PGE(2) synthesis increased in a concentration-dependent manner, showing maximal activity at around 100 nM. S1P treatment also caused the up-regulation of cyclooxygenase-2 (COX-2) mRNA and protein, which was apparent within 3-12 h of stimulation. In terms of the intracellular signaling pathway of S1P-induced WISH cell activation, we found that S1P stimulated two kinds of MAPK, ERK, and p38 kinase. We examined the roles of these two MAPKs in S1P-induced COX-2 expression. S1P-induced COX-2 expression was blocked completely by PD-98059 but not by SB-203580, suggesting that ERK has a critical role in the process. Transfection of S1P(1) or S1P(3) but not of S1P(2) antisense oligonucleotide inhibited S1P-induced COX-2 expression and PGE(2) production in WISH cells, indicating the involvements of S1P(1) and S1P(3) in the processes. This study demonstrates the physiological role of S1P in amniotic fluid and its effect on the modulation of COX-2 expression and PGs synthesis in WISH cells.