Gastrodin alleviates neuronal damage in epileptic cell models by targeting P2RY12 to inhibit microglial hyperactivation

Epilepsy is a chronic neurological disorder characterized by recurrent seizures, with its onset and progression closely linked to neuroinflammation, where abnormal activation and migration of microglia serve as a key pathological process. This study focuses on the regulatory role of gastrodin, the principal active component of the traditional Chinese medicine Gastrodia elata, and its target P2RY12 in chronic epilepsy. Through bioinformatics analysis, P2RY12 was identified as a potential target for Gastrodia elata in treating epilepsy. Molecular docking, Pull-down, and cellular thermal shift assays confirmed that Gastrodin can directly bind to the P2RY12 receptor and inhibit the activation of its downstream RhoA/ROCK signaling pathway. In an in vitro epilepsy model induced by kainic acid (KA), Gastrodin intervention significantly suppressed the migration of microglia toward the injured area and reduced the rearrangement of the F-actin cytoskeleton. Meanwhile, Gastrodin markedly decreased the release of pro-inflammatory cytokines such as TNF-α and IL-1β, alleviated neuronal calcium overload, and inhibited cell apoptosis. Mechanistically, Gastrodin exerts its neuroprotective effects against epilepsy by targeting the P2RY12 receptor to inhibit its mediated chemotactic signaling pathway and inflammatory response, thereby reducing epilepsy-related neuronal damage. Notably, the protective effects of Gastrodin were further enhanced after P2RY12 expression was interfered with, further confirming the critical role of P2RY12 in its neuroprotective action. This study reveals, at the receptor-signaling axis level, the molecular mechanism by which Gastrodin regulates microglial function through P2RY12 to exert anti-epileptic effects, providing not only a solid scientific basis for the clinical application of the traditional Chinese medicine Gastrodia elata but also a novel potential strategy for the treatment of chronic epilepsy.