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[一种改进的大鼠心肌去甲肾上腺素水平微透析研究方法]

[An improved method for microdialysis study of noradrenaline level in rat myocardium].

作者信息

Gilinskiĭ M A, Faĭbushevich A A, Lunte C E

机构信息

Institute of Physiology, Russian Academy of Sciences, Novosibirsk, Russia.

出版信息

Vopr Med Khim. 1998 Jul-Aug;44(4):405-11.

PMID:9845931
Abstract

A level of myocardial norepinephrine (NE) is considered as one of the meaningful parameters in the estimation of myocardium functioning. Long lasting changes of myocardial NE appear to be not only a sequence of pathologic processes in myocardium, but could also be a factor responsible for some diseases. An improved microdialysis sampling technique with HPLC-ED analysis was developed to measure in vivo NE content in a rat myocardial interstitium. Linear polyacrylonitryl 5 mm probes were flushed with methanol while being immersed in water. In vitro NE recovery with the flow rate of perfusate was found at the level 67.9 + 0.4%. Probes were implanted into the rat myocardium under the general anaesthesia. Myocardial dialysate was collected during 20 min into plastic vials, containing 10 microliters of 0.1 M perchloric acid. Low noise, high sensitivity dual electrochemical detection was used for the determination of NE amount in the samples. Overall sensitivity threshold was found about 0.3 pg of NE in 20 microliters. Steady state concentration of NE in myocardial dialysate was found 54 + 7 pg/ml. 20 min before cardiac arrest the concentration of NE in myocardial dialysate was found to be increased 4-5 times. 40 min after cardiac arrest further increase in NE content was registered up to 1-4 ng per ml of perfusate. It is suggested that the method will be useful for routine study of myocardial NE.

摘要

心肌去甲肾上腺素(NE)水平被认为是评估心肌功能的重要参数之一。心肌NE的长期变化似乎不仅是心肌病理过程的一个序列,而且可能是某些疾病的一个致病因素。开发了一种改进的微透析采样技术结合高效液相色谱 - 电化学检测法来测量大鼠心肌间质中的体内NE含量。将5毫米线性聚丙烯腈探针浸入水中时用甲醇冲洗。发现灌注液流速下的体外NE回收率为67.9 + 0.4%。在全身麻醉下将探针植入大鼠心肌。在20分钟内将心肌透析液收集到含有10微升0.1 M高氯酸的塑料小瓶中。采用低噪声、高灵敏度的双电化学检测法测定样品中的NE含量。发现总体灵敏度阈值约为20微升中0.3皮克的NE。发现心肌透析液中NE的稳态浓度为54 + 7皮克/毫升。心脏骤停前20分钟,发现心肌透析液中NE浓度增加了4 - 5倍。心脏骤停后40分钟,NE含量进一步增加,达到每毫升灌注液1 - 4纳克。建议该方法将有助于心肌NE的常规研究。

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