Deparment of Anesthesia Resuscitation Room, Zhongshan Hospital Xiamen University, Xiamen, Fujian, 361004, China.
Deparment of Surgical Anesthesiology, Zhongshan Hospital Xiamen University, Xiamen, Fujian, 361004, China.
Neurotox Res. 2021 Dec;39(6):2007-2017. doi: 10.1007/s12640-021-00427-w. Epub 2021 Oct 15.
Bupivacaine (BUP) is a long-acting amide local anesthetic that may induce strong neurotoxicity and neurological complications. In this study, we elucidate the influence of microRNA-494-3p (miR-494-3p) in BUP-induced neurotoxicity in primary mouse hippocampal neuronal cells. In this study, primary hippocampal neurons were isolated from neonatal C57BL/6 mice. The isolated neurons were treated with various doses of BUP. MTT assay was conducted to analyze neuronal viability. Gene expression measurement was done by RT-qPCR. The impact of miR-494-3p in BUP-mediated neural injury was examined using TUNEL, flow cytometry, western blotting, and ROS activity detection. The regulatory relationship between miR-494-3p and cyclin-dependent kinases 4 and 6 (CDK6) was identified using a luciferase reporter assay. BUP treatment led to neurotoxicity and miR-494-3p upregulation in primary cultured hippocampal neurons. Functionally, miR-494-3p depletion alleviated neuronal apoptosis and oxidative damage induced by BUP. We verified that miR-494-3p targeted and negatively modulated CDK6. MiR-494-3p depletion also activated PI3K/AKT signaling by elevating CDK6 expression in BUP-treated neurons. Furthermore, CDK6 knockdown or PI3K/AKT inactivation attenuated the neuroprotective role of miR-494-3p depletion. Silencing miR-494-3p exerts neuroprotective function in hippocampal neuronal cells against BUP-induced injury by the CDK6-PI3K/AKT pathway.
布比卡因（BUP）是一种长效酰胺类局部麻醉药，可能会引起强烈的神经毒性和神经并发症。在这项研究中，我们阐明了 microRNA-494-3p（miR-494-3p）在原代小鼠海马神经元细胞中 BUP 诱导的神经毒性中的影响。在这项研究中，我们从新生 C57BL/6 小鼠中分离出原代海马神经元。将分离出的神经元用不同剂量的 BUP 处理。通过 MTT 分析测定神经元活力。通过 RT-qPCR 进行基因表达测量。使用 TUNEL、流式细胞术、Western blot 和 ROS 活性检测来检查 miR-494-3p 在 BUP 介导的神经损伤中的作用。通过荧光素酶报告基因检测鉴定 miR-494-3p 与细胞周期蛋白依赖性激酶 4 和 6（CDK6）之间的调节关系。BUP 处理导致原代培养海马神经元的神经毒性和 miR-494-3p 上调。功能上，miR-494-3p 耗竭减轻了 BUP 诱导的神经元凋亡和氧化损伤。我们验证了 miR-494-3p 靶向并负调控 CDK6。在 BUP 处理的神经元中，miR-494-3p 耗竭还通过上调 CDK6 表达激活了 PI3K/AKT 信号通路。此外，CDK6 敲低或 PI3K/AKT 失活减弱了 miR-494-3p 耗竭的神经保护作用。沉默 miR-494-3p 通过 CDK6-PI3K/AKT 通路发挥对 BUP 诱导的海马神经元细胞损伤的神经保护作用。
