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目的以体外原代培养海马神经元网络为模型,应用钙成像技术,观察不同浓度乙酰胆碱(ACh)对海马神经元网络同步自发钙振荡的调控作用。方法取(18±1)d的SD胎鼠,分离海马组织,制成细胞悬液,进行细胞培养。培养9～13d的海马神经元用于实验。分别加入不同浓度的ACh,应用钙成像技术,观察其对钙振荡频率的影响。结果加入1μmol/L ACh后,钙振荡频率增加至对照期的(116.10±2.63)%((?)±s,n=16);加入10μmol/L,100μmol/L ACh后,钙震荡频率增加到对照期的(138.39±4.03)%(n=27)和(141.37±2.33)%(n=21);而加入500μmol/L ACh钙震荡频率在前4min可增加到对照期的(156.81±1.65)%(n=18),但在后6min时段内海马神经元网络同步自发钙震荡频率减慢,到16～18min,钙震荡频率减慢至对照期的(52.27±0.55)%(n=18)。结论 ACh在一定浓度范围内可以加快海马神经元网络同步自发钙振荡频率,调节神经元网络兴奋性。
OBJECTIVE: To investigate the regulatory effect of different concentrations of acetylcholine (ACh) on synchronous spontaneous calcium oscillations in hippocampal neurons using calcium imaging technique in vitro. Methods The fetus of SD fetus (18 ± 1) d was taken out and the hippocampus tissue was separated to prepare cell suspension for cell culture. Cultured hippocampal neurons from 9 to 13 days were used in the experiment. Respectively, adding different concentrations of ACh, the application of calcium imaging technology to observe the impact of calcium oscillation frequency. Results The frequency of calcium oscillations increased to (116.10 ± 2.63)% ((±) ± s, n = 16) after adding 1μmol / L ACh. After adding 10μmol / L and 100μmol / L ACh, (138.39 ± 4.03)% (n = 27) and (141.37 ± 2.33)% (n = 21) in the control period; while the frequency of 500μmol / L ACh calcium oscillation increased to (156.81 ± 1.65) (n = 18). However, the frequency of spontaneous calcium oscillations in the hippocampal neurons network slowed down in the last 6 minutes, and the frequency of calcium oscillations slowed to (52.27 ± 0.55)% (n = 18) . Conclusion ACh can accelerate the synchronization of spontaneous calcium oscillation frequency and regulate the excitability of neuronal networks in a certain concentration range.