目的 制备大鼠脊髓突触体，并对大鼠脊髓突触体中多种神经递质进行定性和定量分析，探究其生理活性。方法 采用不连续性Percoll密度梯度离心法制备大鼠脊髓突触体，并在透射电镜下观察其形态和结构。采用超高效液相色谱-质谱联用法对大鼠脊髓突触体中神经递质的性质及含量进行测定。结果 本实验获得的大鼠脊髓突触体，呈一形态结构完整且具有膜结构的椭圆形颗粒，其中包含有突触小泡和线粒体等。检测出大鼠脊髓突触体中含有多巴胺、去甲肾上腺素、5-羟色胺、高香草酸、3,4-二羟基苯乙酸、5-羟基吲哚乙酸、3-甲氧酪胺、色氨酸、γ-氨基丁酸、犬尿氨酸、3-羟基犬尿氨酸、3-羟基-2-氨基苯甲酸等神经递质，并检测出相应含量，得出相应的线性关系和相关系数。结论 不连续性Percoll密度梯度离心法能制备出大鼠脊髓突触体，并可检测出其中所含神经递质及其含量。
Objective To prepare rat spinal synaptosomes, and to conduct qualitative and quantitative analysis of various neurotransmitters in rat spinal synaptosomes to explore the physiological activities.Methods Rat spinal synaptosomes were prepared by discontinuous Percoll density gradient centrifugation and their morphology and structure were observed under transmission electron microscopy. The properties and content of neurotransmitters in rat spinal synaptosomes were determined by ultra performance liquid chromatography-tandem mass spectrometry.Results Under the experimental conditions, rat spinal synaptosomes were found to contain dopamine, norepinephrine, serotonin, homovanillic acid, 3,4-dihydroxybenzene, 5-hydroxyindoleacetic acid, methoxytyramine, tryptophan, γ-aminobutyric acid, kynurenine, 3-hydroxy kynurenine and 3-hydroxy-2-aminobenzoic acid, and so on. The corresponding neurotransmitter contents were detected, and the corresponding linear relationship and correlation coefficient were obtained.Conclusion Discontinuous Percoll density gradient centrifugation could prepare rat spinal synaptosomes and detect neurotransmitters and their contents in rat spinal synaptosomes.
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