Cheng F C, Yang L L, Yan D Y, Tsai T H, Lee C W, Chen S H
Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.
J Chromatogr A. 2000 Feb 18;870(1-2):389-94. doi: 10.1016/s0021-9673(99)00905-x.
In vivo microdialysis coupled with liquid chromatography was developed for the continuous monitoring of brain neurochemicals during cerebral ischemia in awake, free moving gerbils. The dead volume of the microdialysis system was estimated to be less than 30 microl. The detection limits of the present assay were 0.2 to 2.0 microM for analytes at a signal to noise ratio of five. To validate this assay, a focal cerebral ischemia was produced by occlusion of one common carotid artery for 60 min and then reperfusion for additional 3 h in awake gerbils. A microdialysis probe was inserted into the striatum of the gerbil. Dialysates were autoinjected and analyzed extracellular pyruvate, lactate, and ascorbic acid by liquid chromatography with a UV detector during cerebral ischemia. Significant changes of pyruvate and the lactate/pyruvate ratio were observed. Biphasic and dynamic changes in ascorbic acid and lactate were proposed to correlate a secondary damage. This assay can be used as a tool to study dynamic changes of brain neurochemicals in awake animals.
体内微透析结合液相色谱法被用于在清醒、自由活动的沙鼠脑缺血期间连续监测脑神经化学物质。微透析系统的死体积估计小于30微升。本测定法在信噪比为5时对分析物的检测限为0.2至2.0微摩尔。为验证该测定法,通过结扎一条颈总动脉60分钟,然后在清醒沙鼠中再灌注3小时来产生局灶性脑缺血。将微透析探针插入沙鼠的纹状体。在脑缺血期间,通过带有紫外检测器的液相色谱法自动进样并分析透析液中的细胞外丙酮酸、乳酸和抗坏血酸。观察到丙酮酸和乳酸/丙酮酸比值有显著变化。抗坏血酸和乳酸的双相动态变化被认为与继发性损伤相关。该测定法可作为研究清醒动物脑神经化学物质动态变化的工具。