Growing evidence has suggested the feasibility and effectiveness of blood coagulation with low temperature plasma (LTP) at atmospheric pressure. In the present study we examined morphological changes and contents released out from platelet after LTP treatment to investigate the mechanism of LTP accelerate blood coagulation. Changes of isolated rat platelets treated with LTP had been detected by flow cytometry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Additionally, a scheme that the mixture of different color fluorescent stained platelets treated by LTP and then detected by flow cytometry had proposed to quantitatively assess plasma triggered platelets aggregation. Furthermore, we had used label free quantitative mass spectrometry to identify changes in the supernatant proteome released from LTP treated platelets. On the surface of LTP treatment hastened whole rat blood clots, a layer of semi-transparent film had formed, which had been demonstrated to form from platelet-like membrane structure by ultrastructural analysis with TEM. Flow cytometry combined with fluorescent staining confirmed that LTP treatment promoted platelet aggregation. The observation of plasma treated stained platelets under fluorescence confocal microscopy also confirmed this judgment. We separated the supernatant from the platelets treated with LTP, and then analyzed their differences using mass spectrometry. Compared with control group, Gene ontology (GO) analysis showed that the concentration of 16 protein molecules in the experimental group was increased. LTP treatment can promotes platelet aggregation and the release of platelet activating proteins, which is the potential mechanism of speeding up blood coagulation.

越来越多的证据表明，在大气压下用低温血浆（LTP）进行血液凝结的可行性和有效性。在本研究中，我们检查了LTP治疗后从血小板释放的形态变化和含量，以研究LTP促进血液凝固的机制。通过流式细胞术，透射电子显微镜（TEM）和扫描电子显微镜（SEM）检测了经LTP处理的离体大鼠血小板的变化。另外，提出了一种方案，该方案通过LTP处理然后通过流式细胞术检测不同颜色的荧光染色血小板的混合物，以定量评估血浆触发的血小板聚集。此外，我们使用了无标记定量质谱法来鉴定从LTP处理的血小板释放的上清液蛋白质组中的变化。在LTP处理的表面加速了全大鼠血凝块的形成，形成了一层半透明的膜，通过TEM的超微结构分析已证明该半透明的膜是由血小板状的膜结构形成的。流式细胞仪结合荧光染色证实LTP处理促进血小板聚集。在荧光共聚焦显微镜下观察血浆处理的染色血小板也证实了这一判断。我们从用LTP处理的血小板中分离出上清液，然后使用质谱分析它们的差异。与对照组相比，基因本体论（GO）分析显示，实验组中16个蛋白质分子的浓度增加了。LTP治疗可以促进血小板聚集和血小板活化蛋白的释放，这是加速血液凝固的潜在机制。