权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Regulation of neutrophil functions by platelets

血小板对中性粒细胞功能的调节

基本信息

批准号：
10672174
负责人：
YATOMI Yutaka
金额：
$ 1.86万
依托单位：
Yamanashi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672174/
关键词：
platelet neutrophil aggregation sphingolipids sphingosine sphingosine 1-phosphate ceramide apoptosis スフィンゴシン

项目摘要

(1) We have developed a new device which can sensitively detect the presence of platelet aggregates and platelet-neutrophil aggregates in whole blood. A pulse laser and a CCD camera are attached to a flow cytometer, which is an automated reticulocyte analyzer, R-3000 (Sysmex, Japan). Forward scatter intensity and fluorescence intensity of auramine O, a dye for staining RNA and DNA, are used to selectively gate platelet aggregates and platelet-neutrophil aggregates. Cells that fall in the gated area are checked for their images to correctly distinguish these aggregates from other cell components.Upon stimulation with ADP or serotonin, formation of platelet-neutrophil aggregates was clearly detected with this new device. Now we are trying to develop a system which enables its quantitative measurement.(2) We examined the sphingolipid metabolism of peripheral blood cells, i.e., platelets, erythrocytes, neutrophils, and mononuclear cells. A distinguishing characteristic of sphingolipid metabolism in these highly-differentiated cells was their high sphingosine (Sph) kinase activity. About 40% of platelet Sph 1-phosphate (Sph-1-P) could be released extracellularly by 12-O-tetradecanoylphorbol 13-acetate, possibly through mediation by protein kinase C. On the other hand, in erythrocytes, neutrophils, and mononuclear cells, a significant percentage of Sph-1-P formed inside the cell was discharged without stimulation, while the stimulation-dependent release was marginal. Sph and its methylated derivative, N,N-dimethylsphingosine, induced apoptosis not only in neutrophils but also in mononuclear cells, while Sph-l-P elicited CaィイD22+ィエD2 mobilization in platelets. Our results suggest that all blood cells may remove plasma Sph, which is harmful or suppressive to cellular functions, and change it into Sph-1-P, acting as the source of plasma Sph-1-P, which may play a variety of important roles in blood vessels.

(1)我们研制了一种新的设备，可以灵敏地检测全血中是否存在血小板聚集物和血小板-中性粒细胞聚集物。在一台自动网织红细胞分析仪R-3000(Sysmex，日本)的流式细胞仪上安装了一台脉冲激光和一台CCD摄像机。用于RNA和DNA染色的金胺O的前向散射强度和荧光强度用于选择性地门控血小板聚集物和血小板-中性粒细胞聚集物。检查落在门控区域的细胞的图像，以正确地将这些聚集体与其他细胞成分区分开来。在ADP或5-羟色胺的刺激下，这种新设备可以清楚地检测到血小板-中性粒细胞聚集体的形成。现在我们正在努力开发一种能够进行定量测量的系统。(2)我们检测了外周血细胞，即血小板、红细胞、中性粒细胞和单核细胞的鞘磷脂代谢。在这些高度分化的细胞中，鞘磷脂代谢的一个显著特征是它们具有高的鞘氨醇(Sph)激酶活性。约40%的SPH-1-P可被12-O-十四酰佛波醇13-醋酸酯释放到细胞外，这可能是通过蛋白激酶C介导的。另一方面，在红细胞、中性粒细胞和单核细胞中，有相当大比例的SPH-1-P在细胞内形成的Sph-1-P在没有刺激的情况下释放，而刺激依赖性的释放很少。SPH及其甲基化衍生物N，N-二甲基鞘氨醇不仅可诱导中性粒细胞凋亡，还可诱导单核细胞凋亡，而SPH-L-P可诱导血小板内CaィイD22+ィエD2动员。我们的结果提示，所有的血细胞都可以清除对细胞功能有害或抑制的血浆Sph，并将其转化为Sph-1-P，作为血浆Sph-1-P的来源，可能在血管中发挥多种重要作用。