JunB/AP-1 and NF-κB-mediated induction of nitric oxide synthase by bovine type I collagen in serum-stimulated murine macrophages
JunB/AP-1 and NF-κB-mediated induction of nitric oxide synthase by bovine type I collagen in serum-stimulated murine macrophages
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DOI：
10.1006/niox.2001.0415
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发表时间：
2002-05-01
影响因子：
3.9
通讯作者：
Kim, SG
中科院分区：
文献类型：
--
作者：
Cho, MK;Suh, SH;Kim, SG
I型胶原蛋白占全身胶原蛋白的大部分。特别地，牛I型胶原蛋白被用于医学目的，并作为细胞外基质的纤维组分广泛用于各种细胞培养模型中。本研究旨在探讨I型胶原对血清刺激的Raw 264.7细胞诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）表达的影响及其分子机制。通过北方和西方印迹分析监测，牛I型胶原蛋白增加了血清刺激细胞中NO的产生，而大鼠或小鼠I型胶原蛋白则没有增加，这是由于诱导型一氧化氮合酶的诱导。牛I型胶原蛋白与血清激活的JunB和JunB/AP-1转录复合物的组合，如抗JunB抗体对延迟的AP-1条带的超移和免疫耗竭所证明的。AP-1复合物被抗c-Jun或抗JunD抗体部分免疫耗竭。细胞外信号调节激酶1/2（ERK 1/2），p38激酶，和c-Jun N-末端激酶（JNK）都激活了牛I型胶原在血清刺激的细胞。PD 98059，而不是SB 203580或JNK 1（-）转染，抑制ERK 1/2磷酸化和JunB/AP-1激活。PD 98059或MKK 1（-）转染均抑制iNOS的诱导。iNOS的诱导伴随着NF-κ B的活化和I-κ B α的降解。AP-1和/或NF-κ B诱饵寡核苷酸和吡咯烷二硫代氨基甲酸酯抑制iNOS诱导，这证实AP-1和NF-κ B作为转录因子参与。这些结果表明，牛I型胶原蛋白通过JunB/AP-1和NF-κ B活化诱导血清刺激的小鼠巨噬细胞中的iNOS，并且ERK 1/2的活化在JunB/AP-1活化中起重要作用。(C)2002 Elsevier Science（美国）。
Type I collagen comprises the majority of the total body collagens. In particular, bovine type I collagen is utilized for medical purposes and used widely in a variety of cell culture models as a fibrous component of extracellular matrix. This study was designed to explore the effects of type I collagen on the expression of inducible nitric oxide synthase (iNOS) in serum-stimulated Raw264.7 cells and to study the molecular mechanistic basis. Bovine, but not rat or murine, type I collagen increased NO production in serum-stimulated cells, which resulted from the induction of iNOS, as monitored by Northern and Western blot analyses. Bovine type I collagen in combination with serum activated JunB and JunB/AP-1 transcription complex, as evidenced by supershift and immunodepletion of the retarded AP-1 band with anti-JunB antibody. AP-1 complex was immunodepleted in part by anti-c-Jun or anti-JunD antibody. Extracellular signal-regulated kinase1/2 (ERK1/2), p38 kinase, and c-Jun N-terminal kinase (JNK) were all activated by bovine type I collagen in serum-stimulated cells. PD98059, but not SB203580 or JNK1(-) transfection, inhibited both ERK1/2 phosphorylation and JunB/AP-1 activation. Either PD98059 or MKK1(-) transfection suppressed the iNOS induction. The induction of iNOS accompanied activation of NF-kappaB with degradation of I-kappaBalpha. AP-1 and/or NF-kappaB decoy oligonucleotides and pyrrolidine dithiocarbamate suppressed the iNOS induction, which confirmed involvement of AP-1 and NF-kappaB as transcription factors. These results demonstrated that bovine type I collagen induces iNOS in serum-stimulated murine macrophages through JunB/AP-1 and NF-kappaB activation and that activation of ERK1/2 plays an essential role in JunB/AP-1 activation. (C) 2002 Elsevier Science (USA).