Yingnan Zhao1, Zengkun Li2, Enrong Lu1, Qi Sheng3, Yu Zhao4. 1. Department of Neurology (Six), The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, PR China. 2. Department of Neurology, Harbin First Hospital, Harbin, Heilongjiang 150001, PR China. 3. Department of Neurology, The Third People's Hospital of Shenzhen, Shenzhen, Guangdong 518000, PR China. 4. Department of Neurology (Six), The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, PR China. Electronic address: zhaoyu197309@126.com.
Abstract
OBJECTIVE: Berberine (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. METHODS: In 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. RESULTS: OGD 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. CONCLUSION: BBR protects rats from cerebral I/R injury by up-regulating PPAR-γ to restrain NF-κB-mediated pyroptosis.
OBJECTIVE: Berberine (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. METHODS: In 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. RESULTS: OGD 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. CONCLUSION: BBR protects rats from cerebral I/R injury by up-regulating PPAR-γ to restrain NF-κB-mediated pyroptosis.