Abstract：
BACKGROUND: Previous literature suggests that sphingolipids may impact systemic coagulation and platelet aggregation, thus modulating the risks of thrombotic events. The goal of this investigation was to evaluate the role of serum sphingolipids on intrinsic platelet function to assess whether pharmacologic manipulation of sphingolipid metabolites would impact platelet aggregability.
METHODS: C57BL/6J mice were injected with either normal saline, 1 mg/kg FTY720 (synthetic sphingosine-1-phosphate [S1P] receptor analog), or 5 mg/kg SLM6031434 (sphingosine kinase two inhibitor). Mice were sacrificed at 6 h and whole blood (WB) was collected for impedance aggregometry assessing platelet responsiveness to arachidonic acid or adenosine diphosphate. Ex vivo studies utilized WB or platelet-rich plasma that was pretreated with S1P, FTY720, amitriptyline, or d-sphingosine then analyzed by aggregability and flow cytometry for platelet and platelet-derived microvesicle characteristics.
RESULTS: FTY720 and SLM6031434 pretreated induced similar arachidonic acid and adenosine diphosphate-mediated platelet aggregation as controls. Ex vivo WB and platelet-rich plasma treatment with S1P, FTY720, amitriptyline and d-sphingosine did not impact platelet aggregation. The percentages of CD41+, CD62P+ and CD41+/ceramide+, CD62P+/ceramide + platelets, and platelet-derived microvesicle were not significantly different between amitriptyline-treated and normal saline-treated cohorts.
CONCLUSIONS: Sphingolipid modulating agents, such as FTY720, SLM6031434, S1P, amitriptyline, ceramide, and d-sphingosine do not appear to independently impact platelet aggregation in murine models.
METHODS: C57BL/6J mice were injected with either normal saline, 1 mg/kg FTY720 (synthetic sphingosine-1-phosphate [S1P] receptor analog), or 5 mg/kg SLM6031434 (sphingosine kinase two inhibitor). Mice were sacrificed at 6 h and whole blood (WB) was collected for impedance aggregometry assessing platelet responsiveness to arachidonic acid or adenosine diphosphate. Ex vivo studies utilized WB or platelet-rich plasma that was pretreated with S1P, FTY720, amitriptyline, or d-sphingosine then analyzed by aggregability and flow cytometry for platelet and platelet-derived microvesicle characteristics.
RESULTS: FTY720 and SLM6031434 pretreated induced similar arachidonic acid and adenosine diphosphate-mediated platelet aggregation as controls. Ex vivo WB and platelet-rich plasma treatment with S1P, FTY720, amitriptyline and d-sphingosine did not impact platelet aggregation. The percentages of CD41+, CD62P+ and CD41+/ceramide+, CD62P+/ceramide + platelets, and platelet-derived microvesicle were not significantly different between amitriptyline-treated and normal saline-treated cohorts.
CONCLUSIONS: Sphingolipid modulating agents, such as FTY720, SLM6031434, S1P, amitriptyline, ceramide, and d-sphingosine do not appear to independently impact platelet aggregation in murine models.
摘要：
背景：以前的文献表明，鞘脂可能会影响全身凝血和血小板聚集，从而调节血栓事件的风险。这项研究的目的是评估血清鞘脂对固有血小板功能的作用，以评估鞘脂代谢产物的药理学操作是否会影响血小板聚集性。
方法：C57BL/6J小鼠注射生理盐水，1mg/kgFTY720（合成鞘氨醇-1-磷酸[S1P]受体类似物），或5mg/kg的SLM6031434(鞘氨醇激酶两种抑制剂)。在6h时处死小鼠,收集全血(WB)用于阻抗聚集测定法,评估血小板对花生四烯酸或二磷酸腺苷的反应性。离体研究使用WB或富含血小板的血浆，用S1P预处理，FTY720,阿米替林,或d-鞘氨醇，然后通过聚集性和流式细胞术分析血小板和血小板衍生的微囊泡特征。
结果：FTY720和SLM6031434预处理诱导的花生四烯酸和二磷酸腺苷介导的血小板聚集与对照相似。体外WB和富含血小板的血浆用S1P治疗，FTY720、阿米替林和d-鞘氨醇不影响血小板聚集。CD41+的百分比，CD62P+和CD41+/神经酰胺+，CD62P+/神经酰胺+血小板，和血小板来源的微囊泡在阿米替林治疗组和生理盐水治疗组之间没有显著差异.
结论：鞘脂调节剂，如FTY720，SLM6031434，S1P，阿米替林,神经酰胺,和d-鞘氨醇似乎不独立影响小鼠模型中的血小板聚集。
方法：C57BL/6J小鼠注射生理盐水，1mg/kgFTY720（合成鞘氨醇-1-磷酸[S1P]受体类似物），或5mg/kg的SLM6031434(鞘氨醇激酶两种抑制剂)。在6h时处死小鼠,收集全血(WB)用于阻抗聚集测定法,评估血小板对花生四烯酸或二磷酸腺苷的反应性。离体研究使用WB或富含血小板的血浆，用S1P预处理，FTY720,阿米替林,或d-鞘氨醇，然后通过聚集性和流式细胞术分析血小板和血小板衍生的微囊泡特征。
结果：FTY720和SLM6031434预处理诱导的花生四烯酸和二磷酸腺苷介导的血小板聚集与对照相似。体外WB和富含血小板的血浆用S1P治疗，FTY720、阿米替林和d-鞘氨醇不影响血小板聚集。CD41+的百分比，CD62P+和CD41+/神经酰胺+，CD62P+/神经酰胺+血小板，和血小板来源的微囊泡在阿米替林治疗组和生理盐水治疗组之间没有显著差异.
结论：鞘脂调节剂，如FTY720，SLM6031434，S1P，阿米替林,神经酰胺,和d-鞘氨醇似乎不独立影响小鼠模型中的血小板聚集。
