The mechanism of CaMK2α-MCU-mitochondrial oxidative stress in bupivacaine-induced neurotoxicity
The mechanism of CaMK2α-MCU-mitochondrial oxidative stress in bupivacaine-induced neurotoxicity
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DOI：
10.1016/j.freeradbiomed.2020.04.002
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发表时间：
2020-05-20
影响因子：
7.4
通讯作者：
Xu, Shiyuan
中科院分区：
文献类型：
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作者：
Liu, Zhongjie;Zhao, Wei;Xu, Shiyuan
ca2 + /钙调蛋白依赖性蛋白激酶？（CaMK2?）是神经元中的一种丝氨酸/苏氨酸蛋白激酶，当它被异常激活时会导致神经元损伤。布比卡因是一种用于局部神经阻滞的局麻药，可通过细胞凋亡损伤引起神经毒性。是否CaMK2？与布比卡因诱导的神经毒性有关，其调控机制尚不清楚。本研究采用布比卡因鞘内注射C57BL/6小鼠建立体内损伤模型，并培养人神经母细胞瘤（SH-SY5Y）细胞建立体外损伤模型。结果表明，布比卡因诱导线粒体氧化应激和神经元凋亡损伤，促进CaMK2？camp反应元件结合蛋白（CREB）和线粒体ca2 +单转运蛋白（MCU）表达升高。此外，它还诱导CaMK2？Thr286位点磷酸化，使CREB的Ser133位点磷酸化，上调MCU的转录表达。抑制CaMK2？-MCU信号与击倒CaMK2？和MCU或抑制剂（KN93和Ru360）显著减轻布比卡因诱导的神经毒性损伤。CaMK2过表达？明显增强以上氧化损伤。用激动剂激活MCU逆转siCaMK2的保护作用。布比卡因诱导的线粒体氧化应激。我们的数据显示CaMK2？- mcu -线粒体氧化应激途径是布比卡因诱导神经毒性的主要机制，抑制上述信号通路可能是治疗布比卡因诱导神经毒性的一种治疗策略。
Ca 2+ /calmodulin dependent protein kinase2? (CaMK2?) 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 CaMK2? is involved in bupivacaine-induced neurotoxicity and it is regulated remains unclear. In this study, bupivacaine was ad- ministered 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 CaMK2? and cAMP-response element binding protein (CREB), and elevated mitochondrial Ca 2+ uniporter (MCU) expression. Furthermore, it induced CaMK2? phosphorylation at Thr286 which phosphorylated CREB at Ser133 and up - regulated MCU transcriptional expression. Inhibition of CaMK2?-MCU signaling with knock -down of CaMK2? and MCU or with inhibitors (KN93 and Ru360) signi ficantly mitigated bupivacaine-induced neurotoxic injury. Over -expression of CaMK2? signi ficantly enhanced above oxidative injury. Activated MCU with agonist (sper- mine) reversed protective e ffect of siCaMK2? on bupivacaine-induced mitochondrial oxidative stress. Our data revealed that CaMK2?-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.