Anderson K M, Petrik D, Mullane K, Harris J E
Department of Biochemistry, Rush Medical College, Chicago, IL.
Prostaglandins Leukot Essent Fatty Acids. 1994 Oct;51(4):271-5. doi: 10.1016/0952-3278(94)90191-0.
5,8,11,14-eicosatetraynoic acid (ETYA), an isomorphic competitive analogue of arachidonic acid, spontaneously generates a chemiluminescence signal detected with a liquid scintillation spectrometer operated at ambient temperature in the out-of-coincidence mode. The intensity of the signal was 10- or more-fold above background, required oxygen for its generation, was inhibited by antioxidants, and approximately doubled in D2O. Arachidonic acid, which contains 4-alkene rather than alkyne bonds did no more than double the chemiluminescent signal above background. When examined at 37 degrees C in a Berthold AutoLumat 958 luminometer, DBA (lucigenin) was required to detect a signal above background. Catalase or peroxidase, and to a lesser extent mannitol or histidine but not superoxide dismutase, strongly diminished the signal intensity. These observations provide a baseline for interpreting the functional and electron microscopic changes produced by ETYA in PC3 prostate and A172 glioblastoma cell lines, consistent with a contribution from oxidative stress associated with free radicals, and the absence of these morphological changes in U937 monoblastoid cells.
5,8,11,14-二十碳四炔酸(ETYA)是花生四烯酸的一种同构竞争性类似物,在室温下以非符合模式运行的液体闪烁光谱仪可检测到它自发产生的化学发光信号。该信号强度比背景高10倍或更多倍,其产生需要氧气,会被抗氧化剂抑制,在重水中强度大约会翻倍。含有4-烯烃而非炔键的花生四烯酸使化学发光信号强度比背景增加不到两倍。在37℃下用贝托尔德自动发光计958检测时,需要光泽精(鲁米诺)才能检测到高于背景的信号。过氧化氢酶或过氧化物酶,以及程度稍轻的甘露醇或组氨酸,但超氧化物歧化酶不会,会强烈降低信号强度。这些观察结果为解释ETYA在PC3前列腺癌细胞系和A172胶质母细胞瘤细胞系中产生的功能和电子显微镜变化提供了基线,这与自由基相关的氧化应激作用一致,且U937单核细胞样细胞中不存在这些形态变化。