N-acetylserotonin inhibits oxidized mitochondrial DNA-induced neuroinflammation by activating the AMPK/PGC-1α/TFAM pathway in neonatal hypoxic-ischemic brain injury model

Abstract

Hypoxic-ischemic encephalopathy (HIE) is a substantial cause of irreversible nerve injury in newborns. As the endogenous precursor of melatonin, N-acetylserotonin (NAS) can cross the blood–brain barrier and exert neuroprotective effects. Evidence shows that NAS exerts a neuroprotective role in ischemic brain injury by inhibiting the mitochondrial death pathway. This study investigates whether NAS can play a neuroprotective role in the HIE model and explores its underlying molecular mechanism. In vivo, postnatal day 7 (P7) Sprague-Dawley rats were used to construct the HIE rat model. Significant improvement in short-term neurobehavioral impairment, cerebral infarction volume decrease, and neuronal injury mitigation was observed with NAS treatment after hypoxic-ischemic (HI) injury. In addition, we validated that NAS suppressed NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome activation and NLRP3 inflammasome-related protein expression in the HIE model. Furthermore, we demonstrate that NAS reduced mitochondrial membrane potential loss, oxidative damage to mitochondrial DNA (mtDNA), cytosolic mtDNA copy number and activation of its downstream cyclic guanosine monophosphate adenylate synthase (cGAS)/ stimulator of interferon genes (STING) pathway. Additionally, we further verified the activation of NAS on the AMP-activated protein kinase (AMPK)/ peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α)/ mitochondrial transcription factor A (TFAM) pathway and found that inhibition of AMPK could increase cytosolic mtDNA copy number and NLRP3 inflammasome-related protein expression while decreasing STING protein expression. Our study demonstrated that NAS inhibits oxidative mtDNA-induced neuroinflammation, which is partially associated with AMPK/PGC-1α/TFAM pathway activation in the HIE model.