Li Kehan, Han Xuechang
Department of Anesthesiology, The First Affiliated Hospital of Henan Science and Technology University, No. 24 Jinghua Road, Jianxi District, 471003, Luoyang, China,
J Mol Neurosci. 2015 May;56(1):122-30. doi: 10.1007/s12031-014-0473-6. Epub 2014 Dec 19.
Lidocaine has been indicated to promote apoptosis and to promote endoplasmic reticulum (ER) stress. However, the mechanism underlining ER stress-mediated apoptosis is unclear. In the present study, we investigated the promotion to ER stress in the lidocaine-induced apoptosis in human neuroblastoma SH-SY5Y cells. Firstly, we confirmed that lidocaine treatment induced apoptosis in SH-SY5Y cells, time-dependently and dose-dependently, via MTT cell viability assay and annexin V/FITC apoptosis detection with a FACScan flow cytometer. And the anti-apoptosis Bcl-2 and Bcl-xL were downregulated, whereas the apoptosis-executive caspase 3 was promoted through Western blot assay and caspase 3 activity assay. Moreover, the ER stress-associated binding immunoglobulin protein (BiP), PKR-like ER kinase (PERK), activating transcription factor 4 (ATF4) and CCAAT/enhancer-binding protein homologous protein (CHOP) were also upregulated at both mRNA and protein levels by lidocaine treatment. On the other hand, downregulation of the ER stress-associated BiP by RNAi method not only blocked the lidocaine-promoted ER stress but also attenuated the lidocaine-induced SH-SY5Y cell apoptosis. In conclusion, the present study confirmed the involvement of ER stress in the lidocaine-induced apoptosis in human neuroblastoma SH-SY5Y cells. Our study provides a better understanding on the mechanism of lidocaine's neurovirulence.
利多卡因已被证实可促进细胞凋亡并引发内质网（ER）应激。然而，内质网应激介导的细胞凋亡背后的机制尚不清楚。在本研究中，我们调查了利多卡因诱导人神经母细胞瘤SH-SY5Y细胞凋亡过程中对内质网应激的促进作用。首先，我们通过MTT细胞活力测定以及使用FACScan流式细胞仪进行膜联蛋白V/FITC凋亡检测，证实利多卡因处理可时间和剂量依赖性地诱导SH-SY5Y细胞凋亡。通过蛋白质免疫印迹分析和半胱天冬酶3活性测定发现，抗凋亡蛋白Bcl-2和Bcl-xL表达下调，而凋亡执行蛋白半胱天冬酶3被激活。此外，利多卡因处理还使内质网应激相关的结合免疫球蛋白蛋白（BiP）、蛋白激酶R样内质网激酶（PERK）、活化转录因子4（ATF4）和CCAAT/增强子结合蛋白同源蛋白（CHOP）在mRNA和蛋白水平均上调。另一方面，通过RNA干扰方法下调内质网应激相关的BiP，不仅阻断了利多卡因促进的内质网应激，还减弱了利多卡因诱导的SH-SY5Y细胞凋亡。总之，本研究证实了内质网应激参与利多卡因诱导的人神经母细胞瘤SH-SY5Y细胞凋亡。我们的研究为更好地理解利多卡因神经毒性的机制提供了依据。
