Masuda M, Komiyama Y, Nishikado H, Kuriki H, Egawa H, Murata K
Department of Clinico-laboratory Medicine, Kansai Medical University, Osaka, Japan.
J Leukoc Biol. 1989 Aug;46(2):169-74. doi: 10.1002/jlb.46.2.169.
Membrane fluidity of rat neutrophils was studied following Escherichia coli inoculation, and characteristic changes were observed. Membrane fluidity was assessed by the excimer-forming lipid technique using pyrenedecanoic acid and flow cytometry and expressed as the fluorescence intensity ratios of excimer and monomer pyrenedecanoic acid (IE/IM ratio). High IE/IM ratios indicated high membrane fluidity. The IE/IM ratio of rat neutrophils (0.50 +/- 0.048) increased after E. coli inoculation, reaching a maximum of almost 1.00 after 10-20 min and then returning to its starting value. Intravenous injection of heat-killed E. coli or E. coli-conditioned culture supernatants into rats induced a rapid increase of IE/IM ratios, which returned to initial levels after 20 min. The effect on membrane fluidity of in vitro neutrophil incubation with E. coli, heat-killed E. coli, or E. coli-conditioned culture supernatants was similar to that observed in vivo. Addition of 5 mM ethylenediaminetetraacetic acid (EDTA) did not affect neutrophil membrane fluidity. Addition of either 5 micrograms/ml cytochalasin B or 10(-5) M colchicine did not directly affect neutrophil membrane fluidity but did block the change observed following incubation with bacteria.
在接种大肠杆菌后,对大鼠中性粒细胞的膜流动性进行了研究,并观察到了特征性变化。使用芘癸酸通过准分子形成脂质技术和流式细胞术评估膜流动性,并表示为准分子和单体芘癸酸的荧光强度比(IE/IM比)。高IE/IM比表明高膜流动性。大鼠中性粒细胞的IE/IM比(0.50±0.048)在接种大肠杆菌后增加,在10 - 20分钟后达到最大值近1.00,然后恢复到初始值。给大鼠静脉注射热灭活的大肠杆菌或大肠杆菌条件培养上清液会导致IE/IM比迅速增加,20分钟后恢复到初始水平。体外将中性粒细胞与大肠杆菌、热灭活的大肠杆菌或大肠杆菌条件培养上清液一起孵育对膜流动性的影响与体内观察到的相似。添加5 mM乙二胺四乙酸(EDTA)不影响中性粒细胞膜流动性。添加5微克/毫升细胞松弛素B或10^(-5) M秋水仙碱不会直接影响中性粒细胞膜流动性,但会阻断与细菌孵育后观察到的变化。