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采用双探针微透析技术同步监测沙土鼠局灶性脑缺血时皮质细胞外葡萄糖、丙酮酸、乳酸和谷氨酸水平

Simultaneous monitoring of extracellular glucose, pyruvate, lactate and glutamate in gerbil cortex during focal cerebral ischemia by dual probe microdialysis.

作者信息

Yang D Y, Tsai T H, Cheng C H, Lee C W, Chen S H, Cheng F C

机构信息

Department of Emergency, Chung-Shan Medical and Dental College, Taichung, Taiwan.

出版信息

J Chromatogr A. 2001 Apr 13;913(1-2):349-54. doi: 10.1016/s0021-9673(00)01033-5.

Abstract

The aim of this study was to monitor dynamic changes in energy-related metabolites in the cortex of gerbils subjected to cerebral ischemia by a dual probe microdialysis technique. Focal cerebral ischemia was produced in anesthetized gerbils by occlusion of the right common carotid artery and the right middle cerebral artery for 60 min. Two microdialysis probes were inserted into both sides of the cortex to simultaneously monitor extracellular glucose, lactate, pyruvate and glutamate. Dynamic and comparative changes in these analytes, on the ipsilateral and contralateral sides of the brain, were simultaneously monitored by liquid chromatography and a microdialysis analyzer. The present study demonstrated decreases in glucose and pyruvate, increases in lactate and glutamate on the ipsilateral side whereas all analytes remain constant on the contralateral side of cortex during cerebral ischemia. In vitro recovery of each microdialysis probe was performed to ensure the quality of experiments. The detection limits of pyruvate, glutamate, lactate and glucose were 0.2, 1.0, 2.0 and 20 microM, respectively. The intra- and inter-assay correlations were less than 5% in standard mixtures and pooled brain dialysates.

摘要

本研究的目的是通过双探针微透析技术监测沙土鼠脑缺血时皮质中能量相关代谢物的动态变化。通过阻断右侧颈总动脉和右侧大脑中动脉60分钟,在麻醉的沙土鼠中产生局灶性脑缺血。将两个微透析探针插入皮质两侧,以同时监测细胞外葡萄糖、乳酸、丙酮酸和谷氨酸。通过液相色谱和微透析分析仪同时监测大脑同侧和对侧这些分析物的动态和对比变化。本研究表明,在脑缺血期间,同侧葡萄糖和丙酮酸减少,乳酸和谷氨酸增加,而皮质对侧的所有分析物保持恒定。对每个微透析探针进行体外回收率测定,以确保实验质量。丙酮酸、谷氨酸、乳酸和葡萄糖的检测限分别为0.2、1.0、2.0和20 microM。在标准混合物和合并的脑透析液中,批内和批间相关性均小于5%。

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