Background

Health concerns associated with the exposure to legacy perfluoro-alkyl substances (PFAS) led to the development of new-generation PFAS, such as C6O4. Here we investigated the possible effects of C6O4 on the platelet’s activation profile, by incubating human platelets from healthy donors with C6O4 at different concentrations and evaluating the effects on activation, production and phenotype of platelets micro-particles (MPV) and aggregation under-flow. Based on the eventual platelet pro-aggregation profile detected, the preventive effect of acetylsalicylic acid (ASA) was also explored.

Methods

Adhesion-induced platelet aggregation of platelet rich plasma (PRP) under flow was evaluated on collagen-coated microchip at a shear stress of 10 Dyne. The turbidimetric method was used to investigate platelet aggregation. Finally, the in vitro generation of pro-coagulant MPV in PRP was evaluated by flow cytometry, as characterized by CD41 and annexin V positive events, under resting conditions and after stimulation with agonists at low shear stress.

Results

The generation of platelet aggregates under flow was significantly increased by the pretreatment of PRP with 100–200 ng/mL C6O4, compared to both the control condition and the experiment performed in presence of ASA. Arachidonic acid (AA), ADP and collagen induced an higher maximal aggregation, at turbidimetric evaluation, when PRP was pretreated with 100–500 ng/mL C6O4. In addition, PRP stimulated with AA also showed a steeper slope of the aggregation curve. The aggregation induced by the tested agonists was almost abolished by ASA. Finally, pretreatment with C6O4 increased the number of MPV in resting conditions and in presence of ADP and TRAP. ASA tended to reduce MPV generation.

Conclusions

Exposure to C6O4 associates with an increased platelet response to agonists, translating into a possible increased risk of cardiovascular events. Pending a further clarification on the toxicokinetics of this compound, our results claim the possible prophylactic use of ASA.

中文翻译：

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C6O4对血小板聚集途径的干扰：新一代全氟烷基物质的提示

背景

与传统全氟烷基物质（PFAS）暴露相关的健康问题导致了新一代PFAS（例如C6O4）的发展。在这里，我们通过将健康供体的人血小板与不同浓度的C6O4一起孵育，并评估了对血小板微粒（MPV）和聚集下流的活化，产生和表型的影响，研究了C6O4对血小板活化特性的可能影响。基于最终检测到的血小板聚集前体，还探讨了乙酰水杨酸（ASA）的预防作用。

方法

在胶原涂覆的微芯片上，在剪切力为10达因的条件下，评估了流动引起的富血小板血浆（PRP）粘附诱导的血小板聚集。用比浊法研究血小板聚集。最后，在静息条件下和在低剪切应力下用激动剂刺激后，通过流式细胞术评估了PRP中促凝MPV的体外生成，以CD41和膜联蛋白V阳性事件为特征。

结果

与对照条件和在ASA存在下进行的实验相比，用100–200 ng / mL C6O4预处理PRP可以显着增加流动下的血小板聚集物的生成。当用100–500 ng / mL C6O4预处理PRP时，比浊法评估时花生四烯酸（AA），ADP和胶原蛋白诱导更高的最大聚集。此外，AA刺激的PRP也显示出较陡的聚集曲线斜率。ASA几乎消除了由测试的激动剂诱导的聚集。最后，在静息条件下以及在ADP和TRAP存在下，用C6O4进行预处理可增加MPV的数量。ASA倾向于减少MPV的产生。

结论

暴露于C6O4与血小板对激动剂的反应增加有关，转化为可能增加的心血管事件风险。在对该化合物的毒性动力学进行进一步澄清之前，我们的研究结果声称可以预防性使用ASA。