大鼠脊髓突触体中生理活性物质分析

doi:10.3969/j.issn.1006-9771.2018.12.008

《中国康复理论与实践》 ›› 2018, Vol. 24 ›› Issue (12): 1398-1402.doi: 10.3969/j.issn.1006-9771.2018.12.008

大鼠脊髓突触体中生理活性物质分析

刘正^1,2, 袁文华^1,2, 赵舒煊^1,2, 沈伟^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

收稿日期:2018-09-13 修回日期:2018-09-30 出版日期:2018-12-25 发布日期:2019-01-04
通讯作者: 周海宇(1971-),男,汉族,主任医师、副教授,硕士生导师,主要研究方向：脊柱脊髓的基础与临床研究。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)

Analysis of Physiologically Active Substances in Rat Spinal Synaptosome

LIU Zheng^{1, 2}, YUAN Wen-hua^{1, 2}, ZHAO Shu-xuan^{1, 2}, SHEN Wei^{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

Received:2018-09-13 Revised:2018-09-30 Published:2018-12-25 Online:2019-01-04
Contact: ZHOU Hai-yu. E-mail: gslzzhy2004@163.com
Supported by:
Gansu Natural Science Foundation (No. 17JR5RA189)

摘要/Abstract

摘要： 目的分析测定大鼠脊髓突触体中生理活性物质氨基酸神经递质、ATP、Ca²⁺等。方法不连续性Percoll密度梯度离心法从Sprague-Dawley雌性大鼠中提取脊髓突触体。采用高效液相色谱仪检测其中氨基酸神经递质,紫外分光光度计检测ATP含量、Na⁺-K⁺-ATP酶活性、Ca²⁺-Mg²⁺-ATP酶活性、总ATP酶活性,荧光分光光度计检测Ca²⁺浓度。结果氨基酸神经递质主要为天冬氨酸、甘氨酸、谷氨酸、γ-氨基丁酸;ATP浓度约414.7461 μmol/mg,总ATP酶活性 > Na⁺-K⁺-ATP酶活性 > Ca²⁺-Mg²⁺-ATP酶活性;Ca²⁺浓度可用吸光度值直接换算。结论大鼠脊髓突触体中含有多种生理活性物质,以上方法能准确地进行检测分析,了解其生理活性。

关键词: 脊髓, 突触体, 氨基酸神经递质, 三磷酸腺苷, 钙离子, 大鼠

Abstract: Objective To analyze the physiologically active substances, named amino acid neurotransmitters, ATP and Ca²⁺ in rat spinal synaptosomes. Methods The spinal synaptosome was extracted by discontinuous Percoll density gradient centrifugation from a female Sprague-Dawley rat. The amino acid neurotransmitters were determined with high performance liquid chromatography, ATP content, Na⁺-K⁺-ATPase activity, Ca²⁺-Mg²⁺-ATPase activity and total ATPase activity with ultraviolet spectrophotometer, and Ca²⁺ concentration with fluorescence spectrophotometer. Results The amino acid neurotransmitters contained mainly aspartic acid, glycine, glutamic acid and r-aminobutyric acid. The concentration of ATP was about 414.7461 μmol/mg, total ATPase activity > Na⁺-K⁺-ATPase activity > Ca²⁺-Mg²⁺-ATPasese activity. The concentration of Ca²⁺ could be calculated from absorbancy directly. Conclusion The rat spinal synaptosome contains a variety of physiologically active substances, which can be accurately analyzed to explore their activities.

Key words: spinal cord, synaptosomes, amino acid neurotransmitters, adenosine triphosphate, calcium ion, rats

中图分类号:

R338.2

刘正, 袁文华, 赵舒煊, 沈伟, 冯晓飞, 周海宇. 大鼠脊髓突触体中生理活性物质分析[J]. 《中国康复理论与实践》, 2018, 24(12): 1398-1402.

LIU Zheng, YUAN Wen-hua, ZHAO Shu-xuan, SHEN Wei, FENG Xiao-fei, ZHOU Hai-yu. Analysis of Physiologically Active Substances in Rat Spinal Synaptosome[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2018, 24(12): 1398-1402.

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大鼠脊髓突触体中生理活性物质分析

Analysis of Physiologically Active Substances in Rat Spinal Synaptosome

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