【摘要】 分别取大鼠脑海马、纹体、皮层组织 ,经匀浆、提取、沉淀、吸附等步骤初步分离乙酰胆碱 (Ach)。再经以Tris 马来酸、四甲基氯化铵、辛基磺酸钠溶液为流动相的反相高效液相色谱(HPLC)进一步分离。分离出的Ach被酶反应柱中的乙酰胆碱酯酶催化转变成胆碱 ,再被胆碱氧化酶催化分解为H2 O2 。用工作电压 + 0 5V电化学检测器检测。在这一实验条件下 ,脑样品中乙酰胆碱与胆碱得到很好分离且无其它杂质干扰。在 2～ 1 0nmol范围内线性关系良好。 8次重复测定的变异系数为 1 3%。 1 0次重复测定样品的回收率 ( 1 0 0 7± 3 9) % ,用这一方法测定不同龄大鼠不同脑区Ach含量 ,结果发现 ,成龄大鼠与幼龄大鼠相比 ,海马、纹体、皮层中Ach含量分别减少了 53%、48%和 1 4%。老龄大鼠与成龄大鼠相比 ,海马、纹体、皮层中Ach含量分别减少 52 %、56%和 2 2 %。与学习记忆相关脑区中Ach含量随龄降低的结果提示这些脑区中Ach含量的多少可以作为研究大鼠衰老的指标 ,从而可用于增强学习记忆的研究更多还原

【Abstract】 Hippocampus,striatum and cortex were used separately for identification of acetylcholine (Ach),three regions of rat brain were separated.After a series of process:homogenizing,extracting,precipitating,absorbing were carried out,a primary separation of Ach was obtained.Using Tris\|maleate,tetramethy ammonium chloride and sodium octyl sulfonate as motive phase,the further purification was completed by reverse-phase HPLC.The eluent Ach was transformed into choline (Ch) by acetylcholine sterase in the reaction column,and then using choline oxidase to convert the choline to stoichiometic H 2O 2.The hydrogen peroxide was measured on a electrochemical dector with potential +0.5 V.Under these experimental conditions,Ach and Ch could be separated very well without contaminated interference.In the range of 2—10 nmol,it gave linear relationships between the amounts and the peak areas.Through 8 times repeated determination the coefficient variation was 1.3%.10 time repetition,the recover was (100.7±3.9)%.By this detection method,the amount of Ach in different regions of different age rat gave clear results.The hippocampus,striatum and cortex of the mature rats compared with that of the young rats,their Ach levels reduced 53%,48% and 14% respectively.These results revealed that the determination of Ach in the learning and memory related brain regions could be used as an indicator for doing the research on aging,learning and memory.更多还原