Receptor-mediated activation of murine peritoneal macrophages by antithrombin III acts as a costimulatory signal for nitric oxide synthesis.
Receptor-mediated activation of murine peritoneal macrophages by antithrombin III acts as a costimulatory signal for nitric oxide synthesis.
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DOI：
10.1006/cimm.1998.1337
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发表时间：
1998-08
影响因子：
4.3
通讯作者：
J. Kwak;S. Y. Park;M. Han;H. S. Lee;M. Sohn;U. Kim;J. Mcgregor;W. Samlowski;C. Yim
中科院分区：
文献类型：
--
作者：
J. Kwak;S. Y. Park;M. Han;H. S. Lee;M. Sohn;U. Kim;J. Mcgregor;W. Samlowski;C. Yim
我们评估了抗凝血酶 III (ATIII)（一种丝氨酸蛋白酶抑制剂 (SERPIN)）对诱导小鼠腹膜巨噬细胞中一氧化氮 (NO) 合成的影响。将巨噬细胞与 ATIII 加干扰素-γ (IFN-γ) 一起孵育，而不是单独使用 ATIII，会以剂量依赖性方式诱导亚硝酸盐积累（NO 的代谢物）。通过蛋白质印迹证实了诱导型一氧化氮合酶亚型的表达。 NO 合成可被 NG-单甲基-L-精氨酸、ATIII 与凝血酶复合物或兔抗人 ATIII 抗血清抑制。向巨噬细胞培养物中添加多粘菌素 B 未能抑制 ATIII/IFN-γ 诱导的 NO 合成（排除脂多糖污染）。 125I-ATIII 以剂量依赖性、特异性和可饱和方式与巨噬细胞结合，Km 约为 7.1 nM。我们的结果表明，ATIII，而不是 ATIII/凝血酶复合物，通过一种新的受体介导机制共同刺激巨噬细胞活化和 NO 合成，这可能表明 SERPIN 在巨噬细胞活化中的作用。
We evaluated the effect of antithrombin III (ATIII), a serine protease inhibitor (SERPIN), on induction of nitric oxide (NO) synthesis in murine peritoneal macrophages. Incubation of macrophages with ATIII plus interferon-gamma (IFN-gamma) but not ATIII alone induced nitrite accumulation (a metabolite of NO) in a dose-dependent manner. Expression of the inducible nitric oxide synthase isoform was confirmed by Western blot. NO synthesis was inhibited by NG-monomethyl-l-arginine, by complexing ATIII with thrombin or by rabbit anti-human ATIII antiserum. Addition of polymyxin B to macrophage cultures failed to inhibit ATIII/IFN-gamma-induced NO synthesis, excluding lipopolysaccharide contamination. 125I-ATIII bound to macrophages in a dose-dependent, specific, and saturable manner, with a Km of approximately 7.1 nM. Our results demonstrate that ATIII, but not ATIII/thrombin complex, acts to costimulate macrophage activation and NO synthesis via a novel receptor mediated mechanism, which may indicate a role for SERPINs in macrophage activation.