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

Aims: Bupivacaine, 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 methods: Cell 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 findings: Bupivacaine 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.

Significance: Capillarisin 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.