Illicit Drug N-Ethylpentylone Potentiates Platelet Activation through 5-Hydroxytryptamine2A Receptor-Mediated MAPK Signaling.
Several illicit drugs have been reported to be associated with ischemic stroke and myocardial infarction. However, the underlying mechanisms remain poorly investigated. In this study, we focused on N-Ethylpentylone (NEP), a synthetic cathinone with potent hallucinogenic property and explored its role in platelet activation.
Platelet function assays and flow cytometry were used to evaluate the effects of NEP on platelet aggregation, spreading, clot retraction, and integrin αIIbβ3 activation. The role of the 5-Hydroxytryptamine2A receptor (5-HT2AR) in NEP-mediated platelet responses was investigated using the selective 5-HT2AR antagonist M100907 in both ex vivo and in vivo. Phosphoproteomics identified NEP-regulated phosphorylation sites, and Western blotting validated mitogen-activated protein kinase (MAPK) pathway activation by targeting key phosphorylation nodes.
NEP significantly potentiated platelet aggregation, spreading, clot retraction, and integrin αIIbβ3 activation in a concentration-dependent manner. The NEP potentiated platelet responses were suppressed by M100907 in both ex vivo and in vivo models, confirming the critical regulatory role of 5-HT2AR. Phosphoproteomics revealed NEP-triggered phosphorylation upregulation, enriched in MAPK pathways. Western blotting confirmed selective ERK and p38 phosphorylation, with no effect on JNK.
NEP directly enhances agonist-induced platelet activation by targeting 5-HT2AR on platelet membranes. The findings of this study provide valuable insights for elucidating the hematotoxic mechanisms of NEP and other illicit drugs.
Platelet function assays and flow cytometry were used to evaluate the effects of NEP on platelet aggregation, spreading, clot retraction, and integrin αIIbβ3 activation. The role of the 5-Hydroxytryptamine2A receptor (5-HT2AR) in NEP-mediated platelet responses was investigated using the selective 5-HT2AR antagonist M100907 in both ex vivo and in vivo. Phosphoproteomics identified NEP-regulated phosphorylation sites, and Western blotting validated mitogen-activated protein kinase (MAPK) pathway activation by targeting key phosphorylation nodes.
NEP significantly potentiated platelet aggregation, spreading, clot retraction, and integrin αIIbβ3 activation in a concentration-dependent manner. The NEP potentiated platelet responses were suppressed by M100907 in both ex vivo and in vivo models, confirming the critical regulatory role of 5-HT2AR. Phosphoproteomics revealed NEP-triggered phosphorylation upregulation, enriched in MAPK pathways. Western blotting confirmed selective ERK and p38 phosphorylation, with no effect on JNK.
NEP directly enhances agonist-induced platelet activation by targeting 5-HT2AR on platelet membranes. The findings of this study provide valuable insights for elucidating the hematotoxic mechanisms of NEP and other illicit drugs.
Authors
Cai Cai, Zhang Zhang, Wang Wang, Zhang Zhang, Huang Huang, Jia Jia, Han Han, Zhong Zhong, Wei Wei, Zhang Zhang, Zheng Zheng, Bai Bai, Xu Xu, Hua Hua, Rao Rao
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