Suzuki H, Packham M A, Kinlough-Rathbone R L
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
Exp Mol Pathol. 1995 Feb;62(1):63-72. doi: 10.1006/exmp.1995.1007.
Immunocytochemistry with gold-labeled antibodies was used to compare the effects of stimulation of human platelets with thrombin (1 U/ml) and the thrombin receptor activating peptide, SFLLRN (20 microM). After 3 min, redistribution of fibrinogen, von Willebrand factor, and P-selectin (GMP-140, CD62) was examined, the percentages of [14C]serotonin and beta-thromboglobulin released from pre-labeled platelets were measured, and the amount of thromboxane B2 formed was assayed. Upon stimulation with either thrombin or SFLLRN, the platelets had changed from their normal disc shape to spheroidal forms with short pseudopodia. Few alpha-granules remained, the open canalicular system was expanded (more so with SFLLRN) and contained most of the fibrinogen and von Willebrand factor, although small amounts were evident on the platelet surface. Most of the P-selectin was on the surface. Both thrombin and SFLLRN caused complete release of beta-thromboglobulin and 88.3 and 77.5% release of [14C]serotonin, respectively. However, formation of TXB2 caused by thrombin was 10 times greater than that caused by SFLLRN (969 +/- 173 vs 76 +/- 22 ng/10(9) platelets). Thus, the redistribution of platelet alpha-granule contents is similar with thrombin or SFLLRN stimulation and is unaffected by the extent of thromboxane formation.
采用金标抗体免疫细胞化学法比较凝血酶(1 U/ml)和凝血酶受体激活肽SFLLRN(20 μM)刺激人血小板的效果。3分钟后,检测纤维蛋白原、血管性血友病因子和P-选择素(GMP-140,CD62)的重新分布,测量预标记血小板释放的[14C]5-羟色胺和β-血小板球蛋白的百分比,并测定血栓素B2的生成量。用凝血酶或SFLLRN刺激后,血小板从正常的圆盘状变为带有短伪足的球状。几乎没有α颗粒残留,开放管道系统扩张(SFLLRN刺激时更明显),并包含大部分纤维蛋白原和血管性血友病因子,尽管血小板表面也有少量存在。大部分P-选择素位于表面。凝血酶和SFLLRN均导致β-血小板球蛋白完全释放,[14C]5-羟色胺的释放率分别为88.3%和77.5%。然而,凝血酶引起的TXB2生成量比SFLLRN引起的大10倍(969±173对76±22 ng/10(9)个血小板)。因此,凝血酶或SFLLRN刺激时血小板α颗粒内容物的重新分布相似,且不受血栓素生成程度的影响。