The mechanism of CaMK2α-MCU-mitochondrial oxidative stress in bupivacaine-induced neurotoxicity

Abstract

Ca2+/calmodulin dependent protein kinase2α (CaMK2α) is a serine/threonine protein kinase in neurons and leads to neuronal injury when it is activated abnormally. Bupivacaine, a local anesthetic commonly used in regional nerve block, could induce neurotoxicity via apoptotic injury. Whether or not CaMK2α is involved in bupivacaine-induced neurotoxicity and it is regulated remains unclear. In this study, bupivacaine was administered for intrathecal injection in C57BL/6 mice for building vivo injury model and was used to culture human neuroblastoma (SH-SY5Y) cells for building vitro injury model. The results showed that bupivacaine induced mitochondrial oxidative stress and neurons apoptotic injury, promoted phosphorylation of CaMK2α and cAMP-response element binding protein (CREB), and elevated mitochondrial Ca2+ uniporter (MCU) expression. Furthermore, it induced CaMK2α phosphorylation at Thr286 which phosphorylated CREB at Ser133 and up-regulated MCU transcriptional expression. Inhibition of CaMK2α-MCU signaling with knock-down of CaMK2α and MCU or with inhibitors (KN93 and Ru360) significantly mitigated bupivacaine-induced neurotoxic injury. Over-expression of CaMK2α significantly enhanced above oxidative injury. Activated MCU with agonist (spermine) reversed protective effect of siCaMK2α on bupivacaine-induced mitochondrial oxidative stress. Our data revealed that CaMK2α-MCU-mitochondrial oxidative stress pathway is a major mechanism whereby bupivacaine induces neurotoxicity and inhibition of above signaling could be a therapeutic strategy in the treatment of bupivacaine-induced neurotoxicity.