【摘要】 目的:探讨胞二磷胆碱(citicoline,CC)对6-OHDA诱导的帕金森体外细胞模型的保护作用及其机理。方法:孕15 d大鼠胚胎中脑原代培养,在培养第6、8及10天,实验组加不同浓度CC(2、1、0.1、0.01和0.001 mmol.L-1),并于第11天加50μmol.L-1的6-OHDA作用0.5 h,制作帕金森病细胞模型;6-OHDA组为原代培养细胞加50μmol.L-1的6-OHDA;对照组为原代培养细胞。培养11 d收集细胞。采用MTT法测细胞活力,通过流式细胞仪,用Fluo3/AM检测细胞内[Ca2+]i及用罗丹明123检测线粒体膜电位(Δψm)。结果:2、1和0.1 mmol.L-1CC组,细胞活力与对照组比较明显增高,并随着浓度的增加而增加,差异具有显著性(P<0.05)。1、0.1、0.01和0.001 mmol.L-1CC+6-OHDA组,细胞活力均高于6-OHDA组,差异有显著性(P<0.01)。1、0.1、0.01、0.001 mmol.L-1CC+6-OHDA组细胞内[Ca2+]i均明显下降,分别为(32.23±1.87)%、(17.09±7.45)%、(21.71±8.89)%及(29.18±4.71)%,与6-OHDA组(49.30±7.62)%相比,差异均有显著性(P<0.01);与对照组(42.40±0.81)%比较明显下降,差异均有显著性(P<0.01)。CC+6-OHDA各组与6-OHDA组比较均使Δψm升高,其中1 mmol.L-1CC+6-OHDA组Δψm高于对照组,差异有显著性(P<0.01)。结论:CC通过保护神经元细胞膜、增加细胞活力、降低细胞内[Ca2+]i以及提高Δψm,发挥其对神经元的保护作用。更多还原

【Abstract】 Objective To study the neuroprotective effects of citicoline（CC） on the toxicity induced by 6-OHDA towards dopaminergic mesencephalic neurons in primary culture-Parkinson′s disease（PD） model in vitro and its mechanism. Methods Mesencephalic neurons in culture were prepared from embryonic 15-day Wistar rats.Cultures were treated for 6,8,10 d with various concentrations of CC(2,1,0.1,0.01 and 0.001 mmol·L^-1).At 11th day,the cultures were co-treated with the toxin 6-OHDA(50 μmol·L^-1) for 0.5 h,the cells were collected.Seven groups were categorized as follows:CC(2,1, 0.1,0.01 and 0.001 mmol·L^-1) +6-OHDA,control and（6-OHDA） group.The cell viability was evaluated with MTT assay.Intracellular free Ca²⁺,[Ca²⁺]i,was labeled by using the fluorescent dye Fluo3-AM and detected by flow cytometer.By measuring the intracellular Rhodamine（123 fluorescence） density with flow cytometer,mitochondrial membrane potential（MMP） was evaluated.（Results Cultures） were treated with 2,1 and 0.1 mmol·L^-1 CC,the viability of cell was increased（P<0.05）.1,0.1,0.01 and 0.001 mmol·L^-1 CC significantly attenuated 6-OHDA-induced neurotoxic effects.As compared with（6-OHDA,） the viability of neurons increased,the mean[Ca²⁺]i was significantly lower in cells treated with CC(1,0.1,0.01 and 0.001 mmol·L^-1) plus 6-OHDA（49.30±7.62）% than that in 6-OHDA group without CC treatment（P<0.01）.The densities of [Ca²⁺]i in CC(1,0.1,0.01 and 0.001 mmol·L^-1) plus 6-OHDA groups were（32.23±1.87）%,（17.09±7.45）%,（21.71±8.89）%,（29.18±4.71）%,respectively,and the density of mean [Ca²⁺]i in 6-OHDA group was（49.30±7.62）%.MMP significantly increased（P<0.01）.（Conclusion CC has） an important effect on dopaminergic cell survival in vitro in a validated model of PD.The neurotoxic effect of 6-OHDA can be reduced by CC and CC can increase the viability of neurons,decrease [Ca²⁺]i,maintain MMP at a relatively higher level.更多还原