Salicylate-induced loose coupling: protonmotive force measurements.
Salicylate-induced loose coupling: protonmotive force measurements.
复制标题
水杨酸盐引起的松耦合：质子动力测量。
DOI：
10.1016/0006-2952(85)90774-9
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发表时间：
1985
影响因子：
5.8
通讯作者：
Eguren,LA
中科院分区：
文献类型：
--
作者：
Haas,R;ParkerJr,WD;Stumpf,D;Eguren,LA
卫生局局长关于水杨酸盐和雷伊综合征的报告[1]强调了人们对阿司匹林在雷伊综合征中的作用的关注。一项前瞻性研究[2]发现，患者服用阿司匹林与雷氏综合征之间有100%的相关性，而在精心匹配的对照组中，这一比例为45%。最近的一篇论文指出了水杨酸盐肝毒性和Reye综合征的组织学相似性[3]。多年来，人们一直知道水杨酸盐对完整的线粒体有影响。早期工作[4]显示了解偶联效应，其他早期研究人员[5]报告水杨酸盐刺激线粒体三磷酸腺苷酶(ATPase)活性。Jeffrey和Smith[6]发现水杨酸盐对线粒体肿胀的抑制作用与2，4-二硝基苯酚相似。然而，最近你[7]发现水杨酸盐会引起线粒体肿胀，这与雷氏综合症患者的情况相似。Miyahara和Karler[8]使用Warburg技术，在从大鼠肝脏和大脑分离的线粒体片段中展示了水杨酸盐的解偶联效应。然而，他们的论文强调了对水杨酸盐效应的解释上的混乱。不同的底物可以看到不同的结果，他们的研究结果，就像许多早期的研究一样，可以用水杨酸盐的ATPase刺激作用来解释，这会导致ADP：O比率降低。最近对大鼠肝脏线粒体的研究表明，在水杨酸盐存在的情况下，对跨内线粒体膜的腺嘌呤核苷酸交换以及解偶联效应有影响。当额外基质浓度约为3 mM时，这两种影响都达到了一半。F1-ATPase抑制剂--寡霉素不能阻断水杨酸盐的解偶联作用。有人认为这种解偶联与二硝基苯酚的解偶联密切相关，尽管水杨酸盐还有其他作用[6]。解决表面解偶联机制的一种方法是在水杨酸盐存在的情况下直接测量跨越线粒体内膜的质子动力。本文描述了这些实验的结果。制备了大鼠肝脏线粒体，并进行了如前所述的极谱分析，进行了以下修改[10]。极谱分析在作者自己设计的384pl恒温控制玻璃微室中进行，带有聚四氟乙烯涂层的Microflex搅拌棒。采用克拉克氧电极(辐射计E5046-0)和PM72PO2型辐射计组件作为极谱系统。腔室
The Surgeon General's report on salicylate and Reye Syndrome [1] emphasizes the concern about the role of aspirin in Reye Syndrome. A prospective study [2] found a 100% correlation between aspirin and Reye Syndrome in patients compared to 45% in carefully matched controls. A recent paper pointed out the histological similarity between salicylate hepatotoxicity and Reye Syndrome [3]. Salicylate has been known for many years to have effects on intact mitochondria. Early work [4] showed an uncoupling effect, and other early researchers [5] reported that salicylate stimulates mitochondrial adenosine triphosphatase (ATPase) activity. Jeffrey and Smith [6] found an inhibition of mitochondrial swelling with salicylates similar to that seen with 2, 4-dinitrophenol. However, more recently You [7] found that salicylate produces swelling of mitochondria similar to that seen in patients with Reye Syndrome. Miyahara and Karler [8], using a Warburg technique, demonstrated an uncoupling effect of salicylate in mitochondrial fragments isolated from rat liver and brain. However, their paper emphasized the confusion in interpretation of salicylate effects. Different results were seen with alternate substrates, and the results of their study, as from many earlier studies, could have been explained by an ATPase-stimulating effect of salicylate which would produce the decreased ADP: O ratios seen. More recent studies in rat liver mitochondria demonstrated an effect on the adenine nucleotide exchange across the inner mitochondrial membrane as well as an uncoupling effect in the presence of salicylate [9]. Both the effects were halfmaximal at about 3mM extra matrix concentration. Oligomycin, an F1 ATPase inhibitor, does not block the uncoupling effect of salicylate. It has been suggested that this uncoupling is closely related to that of dinitrophenol in spite of the other effects of salicylate [6]. One approach to resolving the mechanism of apparent uncoupling is to measure directly the protonmotive force across the inner mitochondrial membrane in the presence of salicylate. This paper describes the results of these experiments.Rat liver mitochondria were prepared and polarographic assay was performed as previously described with the following modifications [10]. Polarographic assays were performed in a 384 pl thermostatically controlled glass microchamber of the authors' own design with a Tefloncoated Microflex stir bar. A Clark oxygen electrode (Radiometer E5046-0) was used with a Radiometer model PM72 pO2 module as a polarographic system. Chamber