大鼠脊髓突触体的制备及神经递质的定性和定量分析

doi:10.3969/j.issn.1006-9771.2019.06.010

《中国康复理论与实践》 ›› 2019, Vol. 25 ›› Issue (6): 670-675.doi: 10.3969/j.issn.1006-9771.2019.06.010

大鼠脊髓突触体的制备及神经递质的定性和定量分析

刘正^1,2, 王静³, 沈伟^1,2, 赵舒煊^1,2, 袁文华^1,2, 冯晓飞^1,2, 周海宇¹

1.兰州大学第二医院骨科，甘肃兰州市 730030;
2.甘肃省骨关节疾病研究重点实验室，甘肃兰州市 730030;
3.解放军第九四三医院特诊科，甘肃武威市 733000

收稿日期:2019-01-02 修回日期:2019-02-01 发布日期:2019-06-25
通讯作者: 周海宇，E-mail: gslzzhy2004@163.com
作者简介:刘正(1993-)，男，汉族，重庆市人，硕士研究生，主要研究方向：脊柱脊髓的基础与临床研究。
基金资助:
甘肃省自然科学基金项目(No. 17JR5RA189)

Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters

LIU Zheng^1,2, WANG Jing³, SHEN Wei^1,2, ZHAO Shu-xuan^1,2, YUAN Wen-hua^1,2, FENG Xiao-fei^1,2, ZHOU Hai-yu¹

1.Department of Orthopedics, Second Hospital of Lanzhou University, Lanzhou, Gansu 730030, China;
2.Key Laboratory of Bone and Joint Diseases of Gansu Province, Lanzhou, Gansu 730030, China;
3.Department of Special Medicine, The 943th Hospital of the Chinese People's Liberation Army, Wuwei, Gansu 733000, China

Received:2019-01-02 Revised:2019-02-01 Online:2019-06-25
Contact: ZHOU Hai-yu. E-mail: gslzzhy2004@163.com
Supported by:
Natural Science Foundation of Gansu Province (No. 17JR5RA189)

摘要/Abstract

摘要： 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。

关键词: 脊髓突触体, 神经递质, 超高效液相色谱, 质谱仪, 大鼠

Abstract: 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.

Key words: spinal synaptosomes, neurotransmitters, ultra performance liquid chromatography, mass spectrometer, rats

中图分类号:

R651.2

刘正, 王静, 沈伟, 赵舒煊, 袁文华, 冯晓飞, 周海宇. 大鼠脊髓突触体的制备及神经递质的定性和定量分析[J]. 《中国康复理论与实践》, 2019, 25(6): 670-675.

LIU Zheng, WANG Jing, SHEN Wei, ZHAO Shu-xuan, YUAN Wen-hua, FENG Xiao-fei, ZHOU Hai-yu. Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(6): 670-675.

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大鼠脊髓突触体的制备及神经递质的定性和定量分析

Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters

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