利多卡因固体脂质纳米粒对人神经细胞的毒性作用

Toxicity of lidocaine solid lipid nanoparticles in human neurons

目的 探讨利多卡因固体脂质纳米粒对人神经细胞的毒性作用.方法 采用高压匀质法制备利多卡因固体脂质纳米粒.离体培养入神经母细胞瘤SH-SY5Y细胞,计数并调整细胞密度为5×105个/ml,以每孔100μl接种于96孔板.采用随机数字表法,将细胞分为10组（n=30）,对照组（C组）：常规培养;不同浓度利多卡因组（L1-4组）：利多卡因终浓度依次为1.000％、0.500％、0.250％、0.125％;不同浓度利多卡因固体脂质纳米粒组（L-SLN1-4组）：利多卡因终浓度依次为1.000％、0.500％、0.250％、0.125％;空白固体脂质纳米粒组（SLN组）.于细胞孵育前（C组于相应时点）、培养或孵育1、12和24 h（T0-3）时分别随机取6孔,采用MTT法检测细胞存活率;于T3时光学显微镜下观察细胞形态.结果 与T0时比较,L1-4组和L-SLN12组孵育各时点、L-SLN3组T2.3时、L-SLN4组T3时细胞存活率降低（P＜0.05）,SLN组和C组细胞存活率差异无统计学意义（P＞0.05）.与T1时比较,L1-4组和L-SLN1-3组T2.3时、L-SLN4组T3时细胞存活率降低（P＜0.05）;与T2时比较,L1-4组和L-SLN1-4组T3时细胞存活率降低（P＜0.05）.与C组比较,L1-4组和L-SLN12组各时点、L-SLN3组T2.3时、L-SLN4组T3时细胞存活率降低（P＜0.05）,SLN组细胞存活率差异无统计学意义（P＞0.05）.与相应利多卡因浓度的L-SLN14组比较,L1-4组各时点细胞存活率降低（P＜0.05）.L-SLN1-4组细胞形态改变较L1-4组明显减轻.结论 利多卡因固体脂质纳米粒对人神经细胞具有毒性作用,但该作用弱于利多卡因溶液.

Objective To investigate the toxicity of lidocaine solid lipid nanoparticles （SLNs） in human neurons.Methods Lidocaine-loaded SLNs were prepared using high pressure homogenization.SHSY5Y cells were cultured in vitro and inoculated on 96-well plates （100 μl/well） at a density of 5× 105 cells/ml.SH-SY5Y cells were randomized into 10 groups （n =30 each） using a random number table：control group （group C）, different concentrations of lidocaine groups （L1-4 groups）, different concentrations of lidocaine SLN groups （L-SLN1-4 groups）, and blank SLN group （group SLN）.The cells were cultured routinely in group C.The cells were incubated with the culture medium containing lidocaine with the final concentrations of 1.000%, 0.500%, 0.250% and 0.125% in L1-4 groups, respectively.In LSLN1-4 groups, the cells were incubated with the culture medium containing lidocaine SLNs with the final concentrations of 1.000%, 0.500%, 0.250% and 0.125% in L1-4 groups, respectively.Before incubation （at the corresponding time points in group C）, and at 1, 12 and 24 h of culture or incubation （T0-3） , 6 wells in each group were selected for measurement of the cell survival rate （using methyl thiazolyl tetrazolium assay）.The cell morphology was examined with optical microscope at T3.Results Compared with that at T0, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in L-SLN3 group, and at T3 in L-SLN4 group （P〈0.05） , and no significant change was found in SLN and C groups （P〉0.05）.The cell survival rate was significantly lower at T2,3 in L1-4 and L-SLN1-3 groups, and at T3 in group L-SLN4 than that at T1, and at T3 in L1-4 and L-SLN1-4 groups than that at T2 （P〈0.05）.Compared with group C, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in group L-SLN3, and at T3 in group L-SLN4 （P〈0.05） , and no significant change was found in group SLN （P〉0.05）.Compared with group L-SLN at the corresponding concentration, the cell survival rate was significantly decreased at each time point in group L1-4 （P〈0.05）.Conclusion Lidocaine SLNs have toxic effect on human neurons, but the effect is weaker than that caused by Iidocaine solution.