Apoptosis induction by different local anaesthetics in a neuroblastoma cell line
Apoptosis induction by different local anaesthetics in a neuroblastoma cell line
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DOI：
10.1093/bja/aep236
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发表时间：
2009-11-01
影响因子：
9.8
通讯作者：
Stevens, M. F.
中科院分区：
文献类型：
--
作者：
Werdehausen, R.;Fazeli, S.;Stevens, M. F.
背景。已知局部麻醉剂在临床相关浓度下会诱导细胞凋亡。迄今为止，尚不清楚局部麻醉药的细胞凋亡效力由什么决定。因此，我们比较了局麻药诱导人神经元细胞凋亡与其理化特性的相关性。方法。将神经母细胞瘤细胞 (SHEP) 与八种局部麻醉剂（两种酯类和六种酰胺类）一起孵育。至少评估了每种局部麻醉剂的五个浓度。孵育24小时后，通过流式细胞仪的膜联蛋白V和7-氨基放线菌素D双染色评估早期凋亡阶段的细胞率和总体细胞死亡率。计算并比较所有局部麻醉药导致半最大神经毒性作用 (LD50) 的浓度。结果。所有局部麻醉剂均具有浓度依赖性神经毒性。所有药物在低浓度下均诱导相似的早期凋亡细胞形成率，而在高浓度下，晚期凋亡或坏死细胞死亡占主导地位。比较不同局麻药的LD50值，细胞凋亡效力从高到低依次为：丁卡因>布比卡因>丙胺卡因=甲哌卡因=罗哌卡因>利多卡因>普鲁卡因=阿替卡因。毒性与辛醇/缓冲液系数以及局部麻醉剂的实验效力相关，但与结构（酯或酰胺类型）无关。结论。所有常用的局部麻醉剂在临床使用浓度下都会诱导神经元凋亡。神经毒性与脂溶性相关，因此与局部麻醉剂的传导阻滞效力相关，但与化学类别（酯/酰胺）无关。
Background. Local anaesthetics are known to induce apoptosis in clinically relevant concentrations. Hitherto, it is unknown what determines the apoptotic potency of local anaesthetics. Therefore, we compared apoptosis induction by local anaesthetics related to their physicochemical properties in human neuronal cells.Methods. Neuroblastoma cells (SHEP) were incubated with eight local anaesthetics, two of the ester and six of the amide types. At least, five concentrations of each local anaesthetic were evaluated. After incubation for 24 h, rates of cells in early apoptotic stages and overall cell death were evaluated by annexin V and 7-amino-actinomycin D double staining by flow cytometry. The concentrations that led to half-maximal neurotoxic effects (LD50) were calculated and compared for all local anaesthetics.Results. All local anaesthetics were neurotoxic in a concentration-dependent manner. All drugs induced similar rates of early apoptotic cell formation at low concentrations, whereas at high concentrations, late apoptotic or necrotic cell death predominated. Comparison of LD50 values of the different local anaesthetics resulted in the following order of apoptotic potency from high to low toxicity: tetracaine>bupivacaine>prilocaine=mepivacaine=ropivacaine>lidocaine>procaine=articaine. The toxicity correlated with octanol/buffer coefficients and also with experimental potency of the local anaesthetic, but was unrelated to the structure (ester or amide type).Conclusions. All commonly used local anaesthetics induce neuronal apoptosis in clinically used concentrations. The neurotoxicity correlates with lipid solubility and thus with the conduction blocking potency of the local anaesthetic, but is independent of the chemical class (ester/amide).