Capillarisin protects SH-SY5Y cells against bupivacaine-induced apoptosis via ROS-mediated PI3K/PKB pathway

Abstract

AimsBupivacaine, a common local anesthetic, can induce neurotoxicity and neurological complications. Capillarisin, a bioactive ingredient of Artemisia capillaris root extracts, has been reported to protect SH-SY5Y cells against oxidative stress-mediated neuronal cell death. Nevertheless, the effects of capillarisin on bupivacaine-induced neurotoxicity in SH-SY5Y cells remain unclear. Main methodsCell viability, lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) production, and apoptosis were detected. Malondialdehyde (MDA) content, glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) activities were measured for evaluation of oxidative stress. Western blot was performed to detect the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B (PKB) pathway, and expression of cleaved poly ADP ribose polymerase (PARP), cleaved caspase-3, glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP). Activities of mitochondrial respiratory chain complexes I-III and adenosine triphosphate (ATP) content were measured to evaluate mitochondrial damage. Key findingsBupivacaine treatment dose-dependently reduced cell viability, increased LDH release, and induced ROS production and PI3K/PKB pathway inactivation in SH-SY5Y cells, which were overturned by capillarisin treatment. Capillarisin inhibited bupivacaine-induced apoptosis in SH-SY5Y cells by decreasing cleaved PARP and cleaved caspase-3 expression. Capillarisin inhibited bupivacaine-induced oxidative stress, decrease of mitochondrial respiratory chain complex I, II, and III activities and ATP content, and increase of GRP78 and CHOP expression in SH-SY5Y cells. However, treatment with LY294002 abolished the effects of capillarisin on bupivacaine-induced neurotoxicity in SH-SY5Y cells. SignificanceCapillarisin protected SH-SY5Y cells against bupivacaine-induced apoptosis by inhibiting oxidative stress, mitochondrial injury, and endoplasmic reticulum stress via ROS-mediated of PI3K/PKB pathway.