细胞内、外源活性氧致人多形核白细胞膜理化特性改变的研究

DPH和N-(3芘)马来酰亚胺标记光敏氧化反应及黄嘌呤/黄嘌呤氧化酶反应生成外源性单线态氧(O_2)和超氧阴离子自由基(O_2)作用人多形核白细胞膜脂及膜蛋白质、荧光激发发射光谱形状、峰位未发生改变,荧光强度减小,其中以N-(3芘)马来酰亚胺标记O_2作用的膜蛋白质更为明显.荧光偏振度增大,相应清除剂L-组氨酸、超氧化物歧化酶和过氧化氢酶有抑制效应.调理的酵母多糖刺激中性粒细胞呼吸爆发产生膜、胞内活性氧损伤膜脂、膜蛋白质,测定荧光参数变化与前者不尽相同,DPH荧先强度显著增加,L-组氨酸,超氧化物歧化酶和过氧化氢酶似无抑制效果.

STUDY ON THE ALTERATIONS OF SOME PHYSICOCHEMICAL PROPERTIES IN HUMAN POLYMORPHOUNCLEAR LEUKOCYTE MEMBRANES BY ESTRA-, OR EXTRA- CELLULAR REACTIVE OXYGEN SPECIES

Some alterations of physicochemical properties in human polymorphonuclear leukocyte (PMN) membranes reacted by intra - or extra - cellular generated reactive oxygen species have been investigated by means of fluorescence labels 1, 6 -diphenyl - 1,3, 5hexatriane (DPH) and N - (3 - pyrene) maleimide (N (3p) M) No changes of the peak and shape of flurescence excitation and emission spectra was found, but obvious decrease in the fluorescence intensities and increase in the fluorescence polarizations of both labels in the membranes reacted by extra-cellular generated singlet oxygen (1O2) and superoxide anion (O2 ) were observed. All the above mentioned reactions could be relatively inhibited by L - histidine, superoxide dismutase (SOD) and catalase (CAT) The intracellular generated reactive oxygen species produced by stimulating PMN respiratory burst with opsonized zymosan give rise to the changes of fluorescence parameters of the labels in the reacted membrane being little different to the extracellular generated 1O2 and O2, and moreover the fluorescence intensity of DPH was obviously increased, but the scavenging actions of SOD and CAT on the reactive oxygen species were unlikely to be found.