人类中性粒细胞（PMN）的氧自由基产生与细胞骨架。

Human neutrophil (PMN) oxygen radical production and the cytoskeleton.

作者信息

Wiles M E, Dykens J A, Wright C D

机构信息

Parke-Davis Pharmaceutical Research Division, Warner-Lambert Co., Ann Arbor, MI, USA.

出版信息

Life Sci. 1995;57(16):1533-46. doi: 10.1016/0024-3205(95)02114-x.

DOI:10.1016/0024-3205(95)02114-x

PMID:7564898

Abstract

Electron paramagnetic resonance (EPR) studies were conducted to examine oxygen radical generation following PMN activation by N-formyl-1-methionyl-1-leucyl-1-phenylalanine (fMLP) in the presence or absence of phalloidin and cytochalasin B (CB), agents which stabilize or disrupt f-actin, or taxol and colchicine which stabilize and disrupt microtubule cytoskeletal structures respectively. PMN oxyradical production was monitored using the spin trap 5,5-dimethyl-1-pyrroline n-oxide (DMPO). PMN when unstimulated, treated with phalloidin (10(-6)-10(-8)M), CB (10(-6)-10(-8)M), taxol (10(-6)-10(-8)M), or colchicine (10(-6)-10(-8)M), did not produce a detectable DMPO signal. Stimulation with fMLP (10(-6)M), however, resulted in a significant hydroxyl radical signal which was augmented by PMN treatment with CB (10(-6)-10(-7)M, p < 0.05) and attenuated following PMN treatment with phalloidin (10(-6)-10(-7)M, p < 0.05). Interestingly, colchicine treatment (10(-6)-10(-8)M) significantly attenuated fMLP-mediated oxyradical production, whereas taxol (10(-6)-10(-7)M) significantly increased PMN oxyradical production. These data suggest that stabilization of f-actin and disruption of microtubules attenuates the PMN oxidative burst, whereas disruption of f-actin and stabilization of microtubules increases radical production. These findings suggest cytoskeletal domain-specific contributions to PMN oxidative activity.

摘要

进行了电子顺磁共振（EPR）研究，以检测在存在或不存在鬼笔环肽和细胞松弛素B（CB）（分别稳定或破坏丝状肌动蛋白的试剂）或紫杉醇和秋水仙碱（分别稳定和破坏微管细胞骨架结构的试剂）的情况下，N-甲酰-1-甲硫氨酰-1-亮氨酰-1-苯丙氨酸（fMLP）激活多形核白细胞（PMN）后氧自由基的产生。使用自旋捕获剂5,5-二甲基-1-吡咯啉N-氧化物（DMPO）监测PMN的氧自由基产生。未受刺激的PMN、用鬼笔环肽（10⁻⁶ - 10⁻⁸M）、CB（10⁻⁶ - 10⁻⁸M）、紫杉醇（10⁻⁶ - 10⁻⁸M）或秋水仙碱（10⁻⁶ - 10⁻⁸M）处理时，未产生可检测到的DMPO信号。然而，用fMLP（10⁻⁶M）刺激会产生显著的羟基自由基信号，用CB（10⁻⁶ - 10⁻⁷M，p < 0.05）处理PMN会增强该信号，而用鬼笔环肽（10⁻⁶ - 10⁻⁷M，p < 0.05）处理PMN后该信号会减弱。有趣的是，秋水仙碱处理（10⁻⁶ - 10⁻⁸M）显著减弱fMLP介导的氧自由基产生，而紫杉醇（10⁻⁶ - 10⁻⁷M）显著增加PMN的氧自由基产生。这些数据表明，丝状肌动蛋白的稳定和微管的破坏会减弱PMN的氧化爆发，而丝状肌动蛋白的破坏和微管的稳定会增加自由基产生。这些发现表明细胞骨架区域对PMN氧化活性有特异性贡献。