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。
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.
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