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| 脂肪乳剂减轻布比卡因抑制原代培养海马神经元活力 |
| Lipid emulsion reduces the inhibition of the hippocampus neuronal activity induced by bupivacaine |
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| DOI：10.12089/jca.2018.11.017 |
| 中文关键词: 脂肪乳剂 布比卡因 神经元活力 凋亡 |
| 英文关键词: Lipid emulsion Bupivacaine Neuronal viability Apoptosis |
| 基金项目:国家自然科学基金(81560305) |
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| 中文摘要: |
目的 研究脂肪乳剂对布比卡因致原代培养海马神经元神经毒性的影响。
方法 选择原代培养第8天的海马神经元, 采用四甲基偶氮唑蓝(MTT)法计算布比卡因对海马神经元的半抑制浓度(IC50), 用接近IC50的3个浓度分别作用12 h、24 h和36 h, MTT法检测神经元活力, 观察布比卡因对海马神经元的神经毒性作用, 得出最佳布比卡因浓度和作用时间进行后续实验。将原代海马神经元细胞分为四组(n=3): 空白对照组(C组)不做任何处理; 布比卡因组(B组)加入布比卡因; 脂肪乳剂组(L组)加入1%脂肪乳剂; 布比卡因+脂肪乳剂组(BL组)加入布比卡因和1% 脂肪乳剂。各组均处理一定时间后光学显微镜观察海马神经元生长状态, 检测各组神经元活力, 免疫荧光染色法检测裂解半胱天冬酶-3(cleaved caspase-3)的表达并统计cleaved caspase-3阳性神经元占神经元总数的比例。
结果 布比卡因的IC50为0.033 62%, 摩尔浓度为980.4 μM。选择布比卡因浓度为1 mmol/L和作用时间为24 h进行后续实验。与C组比较, B组海马神经元细胞活力明显减弱(P<0.01)。与B组比较, L组和BL组海马神经元细胞活力明显增强(P<0.01)。与C组比较, B 组和BL组cleaved caspase-3阳性率明显升高(P<0.01), 与B组比较, BL组神经元cleaved caspase-3阳性率明显降低(P<0.01), C组和L组cleaved caspase-3阳性率差异无统计学意义。
结论 布比卡因诱发海马神经元神经毒性呈浓度剂量和时间依赖性, 脂肪乳剂减轻布比卡因的神经毒性可能与增加海马神经元的细胞活力有关。 |
| 英文摘要: |
Objective To study the effect of lipid emulsion on bupivacaine induced neurotoxicity in primary cultured hippocampal neurons.
Methods The eighth day primary cultured hippocampal neurons were used to calculate the half inhibitory concentration (IC50) of bupivacaine by the cytotoxicity using methyl thiazolyl tetrazolium (MTT) assay. Hippocampal neurons were treated with different concentrations close to IC50 for 12 h, 24 h and 36 h detect the neuronal vitality using MTT to observe the neurotoxicity of bupivacaine on hippocampal neurons, then the optimal concentration and treated time of bupivacaine were obtained and used to the subsequent experiments. The primary cultured hippocampal neurons were divided into control group (group C), bupivacaine group (group B), lipid emulsion group (group L), bupivacaine+lipid emulsion group (group BL). Group C without any treatment, group B was treated with bupivacaine, group L was treated with 1% lipid emulsion, group BL was treated with bupivacaine and 1% lipid emulsion. After treatment for a certain period of time, the growth state of hippocampal neurons was observed under light microscopy, the neuronal vitality was detected by MTT and the expression of cleaved caspase-3 was detected using immunofluorescence staining and then the rate of cleaved caspase-3 positive neurons in the total number of neurons was quantified.
Results Bupivacaine had an IC50 of 0.033 62% and a molar concentration of 980.4 μM. The subsequent experiments were performed with a bupivacaine concentration of 1 mmol/L and a duration of 24 h. Compared with group C, the viability of hippocampal neurons in group B was significantly weaker (P<0.01). Compared with group B, the viability of hippocampal neurons in group L and group BL was significantly increased (P<0.01). Compared with group C, the positive rate of cleaved caspase-3 in group B and BL was significantly higher (P<0.01). Compared with group B, the positive rate of cleaved caspase-3 in group BL was significantly lower (P<0.01). There was no significant difference in the positive rate of cleaved caspase-3 between group L and group C.
Conclusion Bupivacaine induced neurotoxicity in hippocampal neurons in a concentration-and time-dependent manner. The lipid emulsion can reduce the neurotoxicity of bupivacaine, which may be related to the increase of cell viability of hippocampal neurons. |
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