## 4 Discussion
Calcium mobilization is another critical factor in clot retraction, as it regulates cytoskeletal rearrangements and platelet contraction [8; 42; 43]. TCE significantly reduced intracellular calcium levels in platelets stimulated by collagen by modulating IP\({}_{3}\)R\({}_{1}\), a channel responsible for calcium release from intracellular stores. This reduction in calcium signaling disrupts cytoskeletal organization, impairing both filopodia retraction and clot volume reduction. Finally, TCE significantly suppressed MAPK (ERK and p38) and PI3K/Akt pathways activated by collagen. These pathways are essential for platelet activation and aggregation, and their inhibition aligns with TCE's effects on suppressing platelet function. The PI3K/Akt and MAPK signaling pathways are known to interact in various pathological conditions, including inflammation-associated thrombosis and cardiovascular diseases [44]. Activation of the PI3K/Akt pathway can lead to the activation of MAPKs, influencing platelet aggregation and thrombus formation. Conversely, MAPK pathways can modulate PI3K/Akt signaling to enhance platelet adhesion and activation, particularly under oxidative stress conditions. The concurrent inhibition of both pathways by TCE suggests that it may exert its anti-platelet effects through multiple interconnected mechanisms rather than acting on a single pathway. Previous studies have demonstrated that Taraxacum species contain bioactive compounds, such as polyphenols and flavonoids, which exhibit anti-inflammatory and anti-thrombotic effects via COX-1 and PI3K/Akt signaling inhibition. This aligns with our findings, suggesting that TCE's bioactive components may contribute to its anti-platelet activity through similar mechanisms. These findings suggest that TCE's effects on platelet signaling pathways may be attributed to its known bioactive constituents and their ability to regulate multiple intracellular signaling cascades involved in platelet function. This study utilized optical aggregometry in platelet-rich plasma (PRP), which does not reflect the effects of erythrocytes and leukocytes on platelet function. PRP was chosen to precisely assess platelet activation mechanisms with minimal interference. However,impedance aggregometry using whole blood could better capture the physiological interactions between these blood components. Future studies should incorporate whole blood-based methods to provide a more comprehensive evaluation of TCE's antiplatelet effects. Although our study provides promising in vitro evidence of TCE's antiplatelet effects, further in vivo studies are needed to assess its efficacy in animal models of thrombosis and hemostasis. Additionally, pharmacokinetic and toxicity evaluations will be essential before considering clinical applications. While flavonoid- and polyphenol-rich extracts, including those from _Taraxacum_ species, have shown potential in cardiovascular research, translating these findings into clinical practice requires comprehensive preclinical validation. In conclusion, TCE effectively inhibits platelet activation and clot retraction by targeting critical pathways, including cAMP-PKA, PIJK/Akt, COX-1, and MAPK signaling. It regulates VASP phosphorylation, suppresses integrin \(\alpha\)IIb\(\beta\)3 activation, and reduces intracellular calcium mobilization, impairing cytoskeletal dynamics necessary for thrombus stabilization. Furthermore, TCE's ability to inhibit COX-1 without inducing hypersensitivity suggests its potential as an alternative antiplatelet agent for individuals intolerant to aspirin. Although this study did not perform a detailed phytochemical analysis, previous research indicates that _T. coreatum_ contains significant levels of flavonoids, such as luteolin and quercetin derivatives, which are known for their antioxidant and anti-inflammatory properties [13]. These compounds may contribute to the observed antiplatelet effects of TCE.