Department of Anesthesiology, BenQ Medical Center, The Affiliated BenQ Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210019, PR China.
Department of Urology and Central Laboratory, BenQ Medical Center, The Affiliated BenQ Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210019, PR China.
J Pharmacol Sci. 2019 Mar;139(3):186-192. doi: 10.1016/j.jphs.2019.01.014. Epub 2019 Feb 19.
Local anesthetics (LAs) may generate neurotoxicity in neurons. In the current study, we explored the mechanisms by which microRNA-132 (miR-132) regulated the neurotoxicity of human neuroblastoma cells (SH-SY5Y) induced by bupivacaine (BUP).
CCK-8, flow cytometry, EdU detection, qRT-PCR and western blotting were used to explore the cell viability, apoptosis and gene expression, respectively.
In this study, we found that 600 μM BUP dramatically inhibited SH-SY5Y cells viability. In addition, BUP induced cell apoptosis and neurotoxicity via increasing active caspase-3 and cleaved PARP1 levels. More importantly, the level of miR-132 was significantly up-regulated in BUP-treated cells, which was significantly reversed by miR-132 inhibitor. In addition, dual-luciferase assay indicated IGF1R was the directly binding target of miR-132 in cells. Our study further indicated that the level of IGF1R was markedly decreased by BUP interference, while miR-132 inhibitor exerted the opposite effect. Furthermore, BUP induced apoptosis and neurotoxicity in SH-SY5Y cells were attenuated by IGF1, which further confirmed IGF1R was the downstream target of BUP in SH-SY5Y cells.
In the present study, miR-132 played important roles in regulating BUP-induced neurotoxicity through IGF1R and may act as a promising molecular target for the treatment of human neurotoxicity induced by BUP.
局部麻醉剂（LAs）可能会导致神经元产生神经毒性。在本研究中，我们探讨了 microRNA-132（miR-132）调节布比卡因（BUP）诱导的人神经母细胞瘤细胞（SH-SY5Y）神经毒性的机制。
CCK-8、流式细胞术、EdU 检测、qRT-PCR 和 Western blot 分别用于探索细胞活力、细胞凋亡和基因表达。
在这项研究中，我们发现 600μM 的 BUP 可显著抑制 SH-SY5Y 细胞活力。此外，BUP 通过增加活性 caspase-3 和裂解 PARP1 水平诱导细胞凋亡和神经毒性。更重要的是，BUP 处理的细胞中 miR-132 的水平显著上调，而 miR-132 抑制剂则显著逆转了这一现象。此外，双荧光素酶报告基因实验表明 IGF1R 是细胞中 miR-132 的直接靶基因。我们的研究进一步表明，BUP 干扰导致 IGF1R 水平明显降低，而 miR-132 抑制剂则产生相反的效果。此外，IGF1 减弱了 BUP 诱导的 SH-SY5Y 细胞凋亡和神经毒性，进一步证实 IGF1R 是 BUP 在 SH-SY5Y 细胞中的下游靶基因。
在本研究中，miR-132 通过 IGF1R 发挥重要作用，调节 BUP 诱导的神经毒性，可能成为治疗 BUP 诱导的人神经毒性的有前途的分子靶点。
