Bone morphogenetic protein‐6 induces the expression of inducible nitric oxide synthase in macrophages

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor‐β (TGF‐β) superfamily. In the present study, we investigated the effect of BMPs on the production of inducible nitric oxide synthase (iNOS) in the murine macrophage cell line, RAW 264.7, and in mouse peritoneal macrophages. Among the BMPs, only BMP‐6 induced iNOS expression in a time‐dependent and dose‐dependent manner in both cell types. Induction of iNOS was inhibited by both cycloheximide and actinomycin D, indicating that the induction of iNOS expression by BMP‐6 requires new protein synthesis. Mechanistic studies revealed that the BMP‐6‐induced iNOS expression requires both Smads and nuclear factor‐kappa B (NF‐κB) signalling pathways. Furthermore, induction of interleukin‐1β (IL‐1β) was necessary for iNOS induction by BMP‐6. These observations suggest that BMP‐6 stimulates macrophages to produce iNOS through IL‐1β via Smad and NF‐κB signalling pathways and that BMP‐6 may be an important regulator of macrophages.

[1]  J. Myklebust,et al.  Inhibitory effects and target genes of bone morphogenetic protein 6 in Jurkat TAg cells , 2007, European journal of immunology.

[2]  Y. Carmi,et al.  The involvement of IL-1 in tumorigenesis, tumor invasiveness, metastasis and tumor-host interactions , 2006, Cancer and Metastasis Reviews.

[3]  J. Massagué,et al.  Smad transcription factors. , 2005, Genes & development.

[4]  K. Miyazono,et al.  BMP receptor signaling: transcriptional targets, regulation of signals, and signaling cross-talk. , 2005, Cytokine & growth factor reviews.

[5]  J. Myklebust,et al.  BMP-6 inhibits growth of mature human B cells; induction of Smad phosphorylation and upregulation of Id1 , 2005, BMC Immunology.

[6]  Zhonghui Liu,et al.  Effects of activin A on the activities of the mouse peritoneal macrophages. , 2005, Cellular & molecular immunology.

[7]  U. Förstermann,et al.  Regulation of the Expression of Inducible Nitric Oxide Synthase , 2003, Biological chemistry.

[8]  M. O’Connor,et al.  Expression of TAK1, a mediator of TGF-beta and BMP signaling, during mouse embryonic development. , 2003, Gene expression patterns : GEP.

[9]  Guang-Quan Zhao,et al.  Consequences of knocking out BMP signaling in the mouse , 2003, Genesis.

[10]  A. Sater,et al.  Regulation of MAP kinase by the BMP-4/TAK1 pathway in Xenopus ectoderm. , 2001, Developmental biology.

[11]  Zhijian J. Chen,et al.  TAK1 is a ubiquitin-dependent kinase of MKK and IKK , 2001, Nature.

[12]  R. Johns,et al.  Hypoxic regulation of inducible nitric oxide synthase via hypoxia inducible factor-1 in cardiac myocytes. , 2000, Circulation research.

[13]  M. Kizaki,et al.  Activin A regulates the production of mature interleukin-1beta and interleukin-1 receptor antagonist in human monocytic cells. , 1998, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[14]  K. Irie,et al.  Role of TAK1 and TAB1 in BMP signaling in early Xenopus development , 1998, The EMBO journal.

[15]  B. Hogan,et al.  Bone morphogenetic proteins: multifunctional regulators of vertebrate development. , 1996, Genes & development.

[16]  A. Kulkarni,et al.  Autoimmune manifestations in the transforming growth factor-beta 1 knockout mouse. , 1996, Blood.

[17]  M. Sporn,et al.  Transforming growth factor beta 1 null mutation in mice causes excessive inflammatory response and early death. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  H. Oppermann,et al.  Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro. , 1992, The Journal of biological chemistry.

[19]  F. Murad,et al.  Induced RAW 264.7 macrophages express soluble and particulate nitric oxide synthase: inhibition by transforming growth factor-beta. , 1992, European journal of pharmacology.

[20]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988, Science.

[21]  M. Sporn,et al.  Deactivation of macrophages by transforming growth factor-β , 1988, Nature.

[22]  H. Shepard,et al.  Inhibition of cytokine production by cyclosporin A and transforming growth factor beta , 1987, The Journal of experimental medicine.

[23]  S. Tannenbaum,et al.  Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. , 1982, Analytical biochemistry.

[24]  M. Sporn,et al.  Transforming growth factors: isolation of polypeptides from virally and chemically transformed cells by acid/ethanol extraction. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Myklebust,et al.  BMP-6 inhibits human bone marrow B lymphopoiesis--upregulation of Id1 and Id3. , 2006, Experimental hematology.

[26]  J. Catravas,et al.  Molecular mechanisms of iNOS induction by IL-1β and IFN-γ in rat aortic smooth muscle cells , 2002 .

[27]  J. Catravas,et al.  Molecular mechanisms of iNOS induction by IL-1 beta and IFN-gamma in rat aortic smooth muscle cells. , 2002, American journal of physiology. Cell physiology.

[28]  M. Sporn,et al.  Deactivation of macrophages by transforming growth factor-beta. , 1988, Nature.