动态监测清醒自由活动大鼠纹状体细胞外液乙酰胆碱和胆碱水平的方法

目的　建立动态监测正常清醒自由活动大鼠纹状体细胞外液中乙酰胆碱 (ACh)和胆碱 (Ch)水平的方法。方法　应用脑内微透析与高效液相色谱 (HPLC ) 柱后固定化酶反应器 (IMER ) 电化学检测法 (ED)相结合的技术 ,动态监测正常清醒自由活动大鼠纹状体细胞外液中ACh和Ch水平。结果　大鼠纹状体细胞外液中ACh和Ch浓度在灌流开始时较高 ,然后逐渐降低 ,ACh在灌流进行 2 4 0min后达到较稳定水平 ;Ch浓度则在灌流的 4 5 0min内持续下降。HPLC IMER ED检测ACh和Ch的极限可达 15 0fmol,且至少在 0 12 5～ 1 0 μmol·L-1的范围内线性关系良好。结论　HPLC IMER ED是一种灵敏且可靠的测定ACh和Ch的方法 ,和微透析技术相结合可以动态监测清醒自由活动大鼠脑内细胞外液中ACh和Ch的水平。

The method of continuous measuring extracellular acetylcholine and choline levels in striatum of freely moving awake rats

AIM To establish the method of continuous measuring extracellular Acetylcholine and Choline levels in[FQ(10*2。46,X-WZ]-striatum of freely moving awake rats. METHODS The microdialysis and high performance liquid chromatography(HPLC)-post column Immobilized enzyme reactor(IMER)-electrochemical detection(ED) were used to continuously measure extracellular acetylcholine and choline levels in striatum of freely moving awake rats. RESULTS Extracellular acetylcholine and choline levels were higher at the beginning of perfusion, then decreased gradually. The ACh concentration reached a stable level after 240 min while the Ch continuously decreased in the whole perfusion period of 450 min. The detection limit for ACh and Ch, at a S/N ratio of three, was found to be 150 fmol per injection. In the range of 0.125～1.0 μmol·L -1 , the relation between the amounts and the peak amplitude was linear. CONCLUSION HPLC-IMER-ED is a sensitive and stable method to measure the ACh and Ch. Coupled with microdialysis, it can been successfully used for monitoring the extracellular ACh and Ch levels of freely moving awake rats. -