Shono M, Yamada M, Yamaguchi H, Miyamoto H
General Laboratories for Medical Research, School of Medicine, University of Tokushima, Japan.
Cell Mol Biol (Noisy-le-grand). 1993 Jun;39(4):361-70.
Fluorescence of peritoneal macrophages, which ingested E. coli labelled with N-[7-(dimethylamino)-4-methyl-coumarinyl]-maleimide, was measured with a microfluorometer. The cellular contents of protein and formazan of a monotetrazolium produced by succinate dehydrogenase (SD) activity were assayed with a microspectrophotometer. During phagocytosis, protein increased in 30 min., but was balanced later by self-hydrolysis. SD activity was unchanged for 30 min. and then increased until 2 hrs. Upon removal of bacteria, the fluorescence and protein decreased with time, whereas SD activity increased further for 1 hr. before decrease. This indicates a delayed activation of the enzyme. The cellular ATP content was reduced during phagocytosis and restored to the normal level 3 hrs. after removal of bacteria. Both KCN-sensitive and -insensitive oxygen uptakes increased in phagocytosis. Thus, oxidative metabolism of macrophages is stimulated in phagocytosis and the stimulation can be demonstrated in situ by the microphotometric methods.
用微量荧光计测量摄取了用N-[7-(二甲基氨基)-4-甲基香豆素基]-马来酰亚胺标记的大肠杆菌的腹膜巨噬细胞的荧光。用显微分光光度计测定由琥珀酸脱氢酶（SD）活性产生的单四氮唑的蛋白质和甲臜的细胞含量。在吞噬作用期间，蛋白质在30分钟内增加，但随后通过自身水解达到平衡。SD活性在30分钟内没有变化，然后增加直到2小时。去除细菌后，荧光和蛋白质随时间下降，而SD活性在下降前进一步增加1小时。这表明酶的激活延迟。细胞ATP含量在吞噬作用期间降低，并在去除细菌后3小时恢复到正常水平。吞噬作用中对KCN敏感和不敏感的氧摄取均增加。因此，巨噬细胞的氧化代谢在吞噬作用中受到刺激，并且可以通过显微光度法在原位证明这种刺激。
