谷氨酸增加大鼠海马脑片可溶性淀粉样前体蛋白的释放

Effect of L-Glutamate on Release of Soluble Amyloid Precursor Protein from Rat Hippocampal Brain Slices

目的 观察兴奋性神经递质谷氨酸对于可溶性淀粉样前体蛋白 (sAPP)释放的影响。方法 雄性SD大鼠断头取海马后 ,于 0～ 4℃Krebs-Ringer液中快速振动切片 ,37℃预孵育后 ,置含谷氨酸或不含谷氨酸Krebs -Ringer液 (对照 )中孵育 ,于 150 0 0g离心 ,取上清 ,用二辛可宁酸法测定总蛋白含量。以Westernblot法测定 ,释放入孵育液中的sAPP。结果 正常孵育的海马脑片能够分泌基础量的sAPP ,谷氨酸刺激能够导致海马脑片释放sAPP增加 ,但没导致新蛋白的产生。结论 谷氨酸在10 -15～ 10 -3 mol·L-1浓度范围内 。

Aim To determine effects of excitatory L-glutamate on the production of the soluble amyloid precursor protein (sAPP).Methods Male Sprague-Dawley rats were decapitated and hippocampus were dissected at 4℃,then hippocampal slices were quickly prepared in the medium of Krebs-Ringer buffer at 0～4℃ by a vibration chopper. After the slices were preincubated in Krebs-Ringer buffer at 37℃,they were stimulated with various concentrations of L-glutamate in Krebs-Ringer buffer.The supernatant was removed from the solution after it centrifugated at 15 000g.Quantities of total protein were measured by bicinchoninic acid assay (BCA).The sAPP derivates released into the incubation medium were assayed by Western blot analysis.Results L-glutamate increased the production of the soluble amyloid precursor protein (sAPP) in incubated hippocampal brain slices,however failed to produce special new protein.Conclusion L-glutamate at the concentrations of 10 -15 mol·L -1 to 10 -3 mol·L -1 have positive effects on the production of the soluble amyloid precursor protein (sAPP).