Tianwang Buxin Decoction ameliorates neuroinflammation in insomnia via adenosine accumulation-mediated AMPK/SIRT1 pathway

ETHNOPHARMACOLOGICAL RELEVANCE: Adenosine (Ado) is a key signaling molecule in the central nervous system. Under cellular stress, extracellular Ado accumulation drives neuroinflammation-induced neuronal damage and sleep dysfunction. Tianwang Buxin Decoction (TWBXD) demonstrates long-term therapeutic efficacy for insomnia. However, the related pharmacological pathways require further mechanistic studies. AIM OF THE STUDY: To elucidate the therapeutic effects of TWBXD against insomnia and its mechanisms in mitigating Ado-induced neuroinflammation. MATERIALS AND METHODS: TWBXD components were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). An in vivo insomnia model was established by inducing adenosine A2A receptor (A2aR) overexpression in the basal forebrain. Sleep architecture was monitored via 24-h polysomnography, followed by analyses of histopathology, Ado levels, A2aR expression, inflammatory mediators, and AMPK/SIRT1 activity. In vitro, neuroinflammation was modeled in a neuron-astrocyte co-culture using A2aR-overexpressing plasmids. TWBXD-containing serum effects on cell viability, Ado dynamics, and inflammatory responses were assessed. RESULTS: Ado accumulation induced neuroinflammation and disrupted sleep. TWBXD exhibited optimal efficacy at 17.6 g/kg compared with other doses. It reversed Ado-mediated suppression of the AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) axis to improve sleep architecture, inhibiting the expression of downstream pro-inflammatory mediators such as nuclear factor-κB, interferon-γ, and interleukin-1β while promoting that of interleukin-10. In vitro experiments and molecular docking verified that TWBXD's neuroprotective effects against Ado overload are mediated by functional A2aR antagonism. CONCLUSION: Neuroinflammation in insomnia is promoted by Ado-driven suppression of the AMPK/SIRT1 signaling pathway. TWBXD restores this axis to alleviate insomnia, representing a novel therapeutic strategy for managing neuroinflammation-related sleep disorders.