Berberine exerts neuroprotective activities against cerebral ischemia/reperfusion injury through up-regulating PPAR-γ to suppress NF-κB-mediated pyroptosis

Abstract

ObjectiveBerberine (BBR) is an anti-inflammatory alkaloid compound extracted from herbs. The purpose of this study is to probe the possible effect and the mechanism of BBR against cerebral ischemia/reperfusion (I/R) injury. MethodsIn vitro oxygen and glucose deprivation (OGD) model was established on neurons from rat hippocampus, which was then subjected to BBR, IVA337 (PPAR-γ agonist), or GW9662 (PPAR-γ antagonist) treatment, to identify their effects on neuronal pyroptosis. MTT assay was utilized to determine cell survival rates, TUNEL staining for observation of β-tubulin and MAP2 expressions, qRT-PCR for detection of mRNA expression of PPAR-γ, Western blot for assessment of protein expressions of PPAR-γ and pyroptosis-related proteins (AIM2, NLPR3, ASC, cleaved-Caspase-1, GSDMD, and GSDMD-N), and ELISA for examination of IL-18 and IL-1β expressions. ResultsOGD modeling induced neuron pyroptosis, as evidenced by increased expression levels of pyroptosis-related proteins as well as IL-1β and IL-18, and elevated cell apoptosis rate. In addition, OGD exposure led to PPAR-γ up-regulation and NF-κB activation. Overexpression of PPAR-γ ameliorated cell pyroptosis, while knockdown of PPAR-γ intensified neuron pyroptosis that could be reversed by BBR. Furthermore, either BBR could block the activation of NF-κB signaling pathway through PPAR-γ. ConclusionBBR protects rats from cerebral I/R injury by up-regulating PPAR-γ to restrain NF-κB-mediated pyroptosis.