Kawa S, Kimura S, Hakomori S, Igarashi Y
The Biomembrane Institute and Department of Pathobiology, University of Washington, Seattle 90104, USA.
FEBS Lett. 1997 Dec 29;420(2-3):196-200. doi: 10.1016/s0014-5793(97)01516-0.
In previous studies, we reported that sphingosine 1-phosphate (Sph-1-P) inhibits the chemotactic motility of some cancer cell lines such as mouse melanoma cells, as well as human smooth muscle cells, at a very low concentration, as demonstrated by a transwell migration assay method (Proc. Natl. Acad. Sci. USA 89, 9698, 1992; J. Cell Biol. 130, 193, 1995). In this study, we investigated the effect of Sph-1-P on the chemotactic motility and invasiveness of human neutrophils, utilizing three different assay systems: (a) a transwell migration assay where IL-8 or fLMP was added as a chemotactic factor, (b) a phagokinetic assay with gold colloids, and (c) a trans-endothelial migration assay with human umbilical vein endothelial cells (HUVECs) plated on collagen layers. We found that among various sphingosine derivatives, Sph-1-P specifically inhibited the IL-8- or fLMP-induced chemotactic migration of neutrophils at concentrations below 1 microM. Phagokinetic activity of neutrophils was also suppressed by Sph-1-P, but more moderately than by the PKC inhibitory sphingosine analog, trimethylsphingosine. Finally, Sph-1-P inhibited trans-endothelial migration and invasiveness of neutrophils into HUVEC-covered collagen layers, whereas no effect on their adhesion to HUVECs was observed. These observations strongly suggest that Sph-1-P can act as a specific and effective motility regulator of human neutrophils, raising the possibility of future applications of Sph-1-P, or its analogs, as anti-inflammatory agents regulating invasive migration of neutrophils through endothelial layers at injured vascular sites.
在先前的研究中，我们报道过1-磷酸鞘氨醇（Sph-1-P）在极低浓度下就能抑制某些癌细胞系（如小鼠黑色素瘤细胞）以及人平滑肌细胞的趋化运动，这是通过Transwell迁移试验方法证实的（《美国国家科学院院刊》89, 9698, 1992；《细胞生物学杂志》130, 193, 1995）。在本研究中，我们利用三种不同的试验系统研究了Sph-1-P对人中性粒细胞趋化运动和侵袭性的影响：（a）添加白细胞介素-8（IL-8）或fLMP作为趋化因子的Transwell迁移试验；（b）用金胶体进行的吞噬动力学试验；（c）用铺在胶原层上的人脐静脉内皮细胞（HUVECs）进行的跨内皮迁移试验。我们发现，在各种鞘氨醇衍生物中，Sph-1-P在浓度低于1微摩尔时能特异性抑制IL-8或fLMP诱导的中性粒细胞趋化迁移。Sph-1-P也抑制中性粒细胞的吞噬动力学活性，但程度比蛋白激酶C（PKC）抑制性鞘氨醇类似物三甲基鞘氨醇要轻。最后，Sph-1-P抑制中性粒细胞向覆盖有HUVECs的胶原层的跨内皮迁移和侵袭，而未观察到其对中性粒细胞与HUVECs黏附的影响。这些观察结果有力地表明，Sph-1-P可作为人中性粒细胞特异性且有效的运动调节因子，这增加了未来将Sph-1-P或其类似物用作抗炎剂的可能性，这类抗炎剂可调节中性粒细胞在受损血管部位穿过内皮层的侵袭性迁移。
