Puerarin suppresses the oxidative stress and activates the cyclic AMP (cAMP)/cAMP response element binding protein/brain-derived neurotrophic factor signaling pathway in alcohol withdrawal-induced depressive disorder via regulating obesity-associated protein–mediated N6-methyladenosine demethylation

OBJECTIVE: Puerarin, a bioactive isoflavone glycoside predominantly extracted from the root of the kudzu plant ( Pueraria lobata ), is a traditional Chinese medicinal herb widely used for centuries. Alcohol withdrawal-induced depression (AWIDD), a serious psychiatric disorder, is prevalent in society. This study aimed to investigate the role of puerarin in oxidative stress and cyclic AMP (cAMP)/cAMP response element binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway in AWIDD, as well as the underlying mechanism. METHODS: An alcohol withdrawal rat model was established. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-px) were measured using commercial kits. The cAMP level was detected by ELISA. CREB and phospho-CREB protein levels were analyzed by Western blot. BDNF level was assessed by reverse transcription-quantitative PCR. Dot blot was used to assess the total N6-methyladenosine (m 6 A) level. The interaction between obesity-associated protein (FTO) and prostaglandin-endoperoxide synthase 1 (PTGS1) was examined through RNA immunoprecipitation and dual-luciferase reporter assays. RESULTS: Puerarin decreased oxidative stress and increased the cAMP, p-CREB, and BDNF levels. Besides, puerarin increased FTO-mediated m 6 A demethylation in alcohol withdrawal rats. FTO inhibition increased oxidative stress and decreased the activation of cAMP/CREB/BDNF signaling pathway. Mechanistically, FTO weakened the stability of PTGS1 mRNA via m 6 A demethylation. Overexpression of PTGS1 reversed the reduction in oxidative stress and the activation of the cAMP/CREB/BDNF signaling pathway induced by FTO overexpression. CONCLUSION: Puerarin suppressed oxidative stress and activated the cAMP/CREB/BDNF signaling pathway in AWIDD via regulating FTO-mediated m 6 A demethylation.