Distinct effect of contraction and ion transport on NADH fluorescence and lactate production in uterine smooth muscle.

G Rubányi, A Tóth, A G Kovách
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Abstract

The present studies were designed to monitor metabolic perturbations following changes in contractile activity and ion transport in rat and rabbit myometrium by simultaneous measurement of NADH fluorescence, lactate production and isometric force. Stimulation (127 mM K+) and inhibition (Ca2+-deficient solution) of isometric force development induced a decrease/increase of fluorescence intensity corresponding to a more oxidized/reduced state of tissue pyridine nucleotides, respectively. If tension development was abolished by EGTA, fluorescence changes due to altered ion transport could be monitored. Slow progressive stimulation of the sodium pump by Ca2++-deficient solution resulted in an ouabain (10(-3) M) sensitive monophasic NADH oxidation, which was reversed by inhibition of the pump by isotonic K+ solution. If, however, stimulation of the pump was rapid and maximal (addition of 30 mM KCl to Na+-loaded tissues) the fluorescence response was triphasic and ouabain sensitive: it consisted of an initial NAD reduction followed by a transient NADH oxidation and a second slow NAD reduction. These fluorescence changes are interpreted in terms of separate redox changes in the cytoplasm and mitochondria. The ouabain sensitive stimulation or inhibition of Na/K transport was always accompanied by parallel changes in uterine lactate production irrespective of the actual contractile state of the myometrium. The present results show that 1 fluorescence technique can be applied to monitor metabolic perturbations in the uterus evoked by changes in contractility or ion transport; and 2 aerobic glycolysis and Na/K transport are tightly coupled in the myometrium.

收缩和离子转运对子宫平滑肌NADH荧光和乳酸生成的明显影响。
本研究旨在通过同时测量NADH荧光、乳酸生成和等长力来监测大鼠和家兔肌层收缩活性和离子转运变化后的代谢扰动。刺激(127 mM K+)和抑制(Ca2+缺乏溶液)的等长力发展诱导荧光强度的降低/增加,分别对应于组织吡啶核苷酸的更多氧化/还原状态。如果EGTA消除了张力发展,则可以监测离子传输改变引起的荧光变化。Ca2++缺乏溶液对钠泵的缓慢进行性刺激导致瓦巴因(10(-3)M)敏感的单相NADH氧化,等渗K+溶液对钠泵的抑制可以逆转这一氧化。然而,如果泵的刺激是快速和最大的(向Na+负载的组织中添加30 mM KCl),则荧光响应是三相的和瓦巴因敏感的:它包括最初的NAD还原,随后是短暂的NADH氧化和第二次缓慢的NAD还原。这些荧光变化被解释为细胞质和线粒体中单独的氧化还原变化。无论子宫肌层的实际收缩状态如何,乌巴因对Na/K转运的敏感刺激或抑制总是伴随着子宫乳酸生成的平行变化。结果表明,1荧光技术可用于监测子宫收缩力或离子转运变化引起的代谢扰动;2有氧糖酵解和Na/K转运在肌层紧密耦合。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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