## 4 Discussion  
Platelets play a critical role in hemostasis; however, their overactivation under pathological conditions can lead to thrombotic diseases such as ischemic stroke, myocardial infarction, and atherosclerosis [35; 36]. This study demonstrates that TCE has potent inhibitory effects on platelet activation and clot retraction by targeting multiple signaling pathways and cellular mechanisms involved in platelet function. TCE effectively inhibited platelet aggregation induced by collagen, thrombin, and ADP, which are mediated through specific membrane receptors. However, TCE had no effect on TPA-induced aggregation, as TPA bypasses membrane receptor-mediated pathways by directly activating PKC. This suggests that TCE's anti-platelet effects are selective for receptor-dependent signaling pathways rather than broadly inhibiting all platelet activation mechanisms. To further explore this, we examined TCE's influence on key intracellular signaling pathways involved in platelet activation. The observed inhibition of granule release, calcium mobilization, and integrin \(\alpha\)Ilb\(\bar{6}3\) activation suggests that TCE primarily modulates upstream pathways such as PI3K/Akt and eicosanoid signaling, which are crucial for receptor-mediated platelet activation. The lack of effect on TPA-induced aggregation  
Figure 6: Modulation of MAPK and PI3K/Akt Signaling Pathways by TCE. The phosphorylation status of key signaling molecules was assessed in platelets pretreated with TCE, followed by stimulation with collagen (2.5 ug/mL). (**A**) Influence of TCE on MAPK signaling: Phosphorylation levels of ERK and p38 were analyzed. (**B**) Effects of TCE on PI3K/Akt pathway components: Changes in phosphorylation of PI3K, Akt, PDK, and GSK3\(\bar{6}\) were evaluated. Data are expressed as mean \(\pm\) SD (\(t\) = 3). Statistical significance is indicated as \({}^{*}p<0.05\) and \({}^{**}p<0.001\) compared to collagen-treated platelets. supports the notion that TCE does not directly interfere with PKC-dependent pathways, highlighting its specificity in modulating platelet function. Additionally, TCE reduced TXA\({}_{2}\) synthesis by inhibiting COX-1 activity, a key enzyme in the eicosanoid pathway. Suppression of TXA\({}_{2}\) synthesis attenuates downstream platelet activation, similar to aspirin, a well-known COX-1 inhibitor [37]. Previous studies have shown that flavonoids and phenolic acids from _Taraxacum_ species exhibit anti-inflammatory and anti-thrombotic effects via COX-1 and PI3K/Akt signaling inhibition [38]. This aligns with our findings, suggesting that TCE's bioactive components may contribute to its anti-platelet activity through similar mechanisms. Importantly, TCE offers potential therapeutic advantages over aspirin for individuals with aspirin hypersensitivity or intolerance, as it does not trigger adverse effects such as aspirin-exacerbated respiratory disease. However, although TCE demonstrates promising antiplatelet effects, this study does not include preclinical toxicity or pharmacokinetic assessments, which are essential for evaluating its therapeutic applicability. Future research should investigate the safety profile, bioavailability, and metabolism of TCE to determine its suitability as an alternative antiplatelet agent. TCE also significantly inhibited clot retraction, a process critical for thrombus stabilization. Clot retraction is mediated by the contraction of platelet filopodia, which requires an intact cytoskeleton comprising actin filaments and microtubules [30; 39]. TCE's inhibition of clot retraction involves multiple mechanisms. First, TCE activates the cAMP-PKA pathway, leading to the phosphorylation of VASP at Ser157, a key site associated with reduced interaction with actin filaments. This disruption impairs actin polymerization and filopodia formation, which are essential for clot retraction [40; 41]. Second, TCE suppresses integrin \(\alpha\)IIb\(\beta\)3 activation, further impairing the contractile forces exerted on fibrin fibers.