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巨噬细胞分泌基质金属蛋白酶的分析

Analysis of matrix metalloproteinase secretion by macrophages.

作者信息

Elkington Paul T, Green Justin A, Friedland Jon S

机构信息

Department of Infectious Diseases & Immunity, Imperial College London, Hammersmith Campus, London, UK.

出版信息

Methods Mol Biol. 2009;531:253-65. doi: 10.1007/978-1-59745-396-7_16.

Abstract

Matrix metalloproteinases (MMPs) are zinc-dependent proteases whose physiological roles include control of leukocyte migration. They are implicated in tissue destruction in inflammatory and infectious diseases. MMPs are not only capable of degrading all components of the extracellular matrix, but they also can modulate the immune response by cleaving cytokines and chemokines to alter their activity. Macrophages secrete a broad range of MMPs and represent a key source of MMPs in inflammatory lesions such as granulomas. Zymography is substrate-based gel electrophoresis that allows direct visualization of MMP activity. Here we describe measurement of MMP secretion from macrophages focusing on quantitative zymography. We also discuss complementary methods that should be used in parallel with zymography. The ability to analyze and quantify MMP secretion by macrophages offers an additional window through which to understand the contributions of macrophages to a wide variety of infectious, inflammatory, and immunologic disorders.

摘要

基质金属蛋白酶(MMPs)是锌依赖性蛋白酶,其生理作用包括控制白细胞迁移。它们与炎症和感染性疾病中的组织破坏有关。MMPs不仅能够降解细胞外基质的所有成分,还可以通过切割细胞因子和趋化因子来改变其活性,从而调节免疫反应。巨噬细胞分泌多种MMPs,是肉芽肿等炎症病变中MMPs的关键来源。酶谱法是一种基于底物的凝胶电泳,可直接观察MMP活性。在此,我们描述了以定量酶谱法为重点的巨噬细胞MMP分泌的测量方法。我们还讨论了应与酶谱法并行使用的补充方法。分析和量化巨噬细胞MMP分泌的能力为理解巨噬细胞对多种感染性、炎症性和免疫性疾病的贡献提供了一个额外的窗口。

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