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目的:研究PAF诱导P-选择素表达的细胞内信号机制。方法:置备人血小板悬液,用流式细胞术测定PAF诱导的P-选择素表达。结果:用依他酸2mmol·L~(-1)和BAPTA200μmol·L~(-1)分别阻断外钙内流和络合内钙,显著抑制PAF诱导的P-选择素表达[13.3±0.9)％;(16.8±1.9)％ vs PAF对照组(47.5±1.3)％,P<0.01]。用阿米洛利400μmol·L~(-1)抑制Na~+/H~+交换,Genistein 300μmol·L~(-1)抑制蛋白酪氨酸磷酸化也均可抑制PAF引起的P-选择素上调[(37.5±2.1)％;(29±4)％ vsPAF对照组(47.5±1.3)％,P<0.01)]。结论:蛋白酪氨酸磷酸化,细胞内钙动员及Na~+/H~+交换激活中介PAF诱导的血小板P-选择素表达。
OBJECTIVE: To study the intracellular signal mechanism of PAF-induced P-selectin expression. Methods: Human platelet suspension was prepared and PAF-induced P-selectin expression was measured by flow cytometry. Results: The intracellular calcium influx and complexation intracellular calcium were blocked by 2mmol·L -1 etoposide and 200 μmol·L -1 BAPTA respectively, and significantly inhibited the P-selectin expression induced by PAF [13.3 ± 0.9 )%; (16.8 ± 1.9)% vs PAF control group (47.5 ± 1.3)%, P <0.01]. Inhibition of Na ~ + / H ~ + exchange by 400μmol·L -1 amiloride inhibited the phosphorylation of tyrosine phosphorylated by genistein 300μmol·L -1, which also inhibited P-selectin (37.5 ± 2.1)%; (29 ± 4)% vs the level of PAF (47.5 ± 1.3)%, P <0.01). CONCLUSION: Protein tyrosine phosphorylation, intracellular calcium mobilization and Na ~ + / H ~ + exchange activate PAF-induced platelet P-selectin expression.