Reactive oxygen species produced by macrophage-derived foam cells regulate the activity of vascular matrix metalloproteinases in vitro - Implications for atherosclerotic plaque stability
Reactive oxygen species produced by macrophage-derived foam cells regulate the activity of vascular matrix metalloproteinases in vitro - Implications for atherosclerotic plaque stability
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DOI：
10.1172/jci119076
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发表时间：
1996-12-01
影响因子：
15.9
通讯作者：
Galis, ZS
中科院分区：
文献类型：
--
作者：
Rajagopalan, S;Meng, XP;Galis, ZS
动脉粥样硬化斑块的易损区域通常含有富含脂质的巨噬细胞，并显示出基质金属蛋白酶活性。我们假设巨噬细胞源性泡沫细胞释放的活性氧物质可触发血管间质中潜在的金属蛋白酶原形式的活化。我们发现，从高胆固醇血症兔体内分离出的体内生成的巨噬细胞泡沫细胞可产生超氧化物、一氧化氮和过氧化氢。我们使用培养的人血管平滑肌细胞分泌的金属蛋白酶在体外测试了这些活性氧以及可能由一氧化氮和超氧化物同时产生的过氧亚硝酸盐的作用。通过SDS - PAGE酶谱分析、蛋白质印迹和酶活性测定法检测培养基中的酶或亲和纯化的（前基质金属蛋白酶 - 2和基质金属蛋白酶 - 9）。在所使用的条件下，与黄嘌呤/黄嘌呤氧化酶一起孵育增加了活性明胶酶的量，而一氧化氮供体没有明显的作用。与过氧亚硝酸盐一起孵育导致基质金属蛋白酶 - 2的硝化，并赋予其胶原蛋白溶解活性。过氧化氢处理显示出一种过氧化氢酶可逆的双相效应（在4μM浓度下明胶酶活化，在≥10 - 50μM浓度下抑制）。因此，活性氧物质可调节高氧化应激区域的基质降解，从而可能导致动脉粥样硬化斑块的不稳定。
Vulnerable areas of atherosclerotic plaques often contain lipid-laden macrophages and display matrix metalloproteinase activity. We hypothesized that reactive oxygen species released by macrophage-derived foam cells could trigger activation of latent proforms of metalloproteinases in the vascular interstitium. We showed that in vivo generated macrophage foam cells produce superoxide, nitric oxide, and hydrogen peroxide after isolation from hypercholesterolemic rabbits. Effects of these reactive oxygens and that of peroxynitrite, likely to result from simultaneous production of nitric oxide and superoxide, were tested in vitro using metalloproteinases secreted by cultured human vascular smooth muscle cells. Enzymes in culture media or affinity-purified (pro-MMP-2 and MMP-9) were examined by SDS-PAGE zymography, Western blotting, and enzymatic assays. Under the conditions used, incubation with xanthine/xanthine oxidase increased the amount of active gelatinases, while nitric oxide donors had no noticeable effect. Incubation with peroxynitrite resulted in nitration of MMP-2 and endowed it with collagenolytic activity. Hydrogen peroxide treatment showed a catalase-reversible biphasic effect (gelatinase activation at concentrations of 4 mu M, inhibition at greater than or equal to 10-50 mu M). Thus, reactive oxygen species can modulate matrix degradation in areas of high oxidant stress and could therefore contribute to instability of atherosclerotic plaques.