Intracellular signaling pathways involved in the release of IL-4 and VEGF from human keratinocytes by activation of kinin B1 receptor: functional relevance to angiogenesis.
The injured skin produces a number of mediators that directly or indirectly modulate cell chemotaxis, migration, proliferation, and angiogenesis. Components of the kinin pathway including the kinin B1 receptor (B1R) have been found to occur in the human skin, but information about its role on keratinocyte biology is still scarce. Our aim was to determine whether stimulation of B1R causes the secretion of IL-4 and/or VEGF from human keratinocytes and to evaluate the role of the B1R agonist Lys-des[Arg(9)]bradykinin and IL-4 on various stages of angiogenesis, such as cell migration, proliferation, and release of metalloproteases. By using ELISA and Western blotting, we showed that HaCaT keratinocytes stimulated with the B1R agonist release IL-4 and VEGF. Stimulation of B1R also caused transient c-JunN-terminal kinase phosphorylation and JunB nuclear translocation, transcription factor that regulates IL-4 expression. The 3D-angiogenesis assay, performed on spheroids of EA.hy923 endothelial cells embedded in a collagen matrix, showed that their cumulative sprout area increased significantly following stimulation with either IL-4 or B1R agonist. Furthermore, these ligands produced significant endothelial cell migration and release of metalloproteases 2 and 9, but did not increase endothelial cell proliferation as measured by 5-bromo-2'-deoxyuridine incorporation. Our results provide experimental evidence that establishes IL-4 and B1R agonist as important angiogenic factors of relevance for skin repair.
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile.
Astrid J
AJ
Mejia
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile.
Carola E
CE
Matus
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile.
Francisca
F
Pavicic
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile.
Miguel
M
Concha
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile.
Pamela
P
Ehrenfeld
Laboratorio de Patologia Celular, Instituto de Anatomia, Histologia y Patologia, Universidad Austral de Chile, Isla Teja, PO Box 567, Valdivia, Chile. cfiguero@uach.cl.
Carlos D
CD
Figueroa
D042501:Angiogenic Proteins / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D042783:Endothelial Cells / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism* / Q000494:pharmacology*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D007609:Kallidin / Q000031:analogs & derivatives / Q000494:pharmacology; D015603:Keratinocytes / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010766:Phosphorylation; D016755:Proto-Oncogene Proteins c-jun; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D043783:Receptor, Bradykinin B1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D012867:Skin / Q000166:cytology / Q000378:metabolism; D018874:Spheroids, Cellular; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D042501:Angiogenic Proteins; D017077:Culture Media, Conditioned; C508598:IL4 protein, human; D016755:Proto-Oncogene Proteins c-jun; D034741:RNA, Small Interfering; D043783:Receptor, Bradykinin B1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D015847:Interleukin-4; D007609:Kallidin; C052787:kallidin, des-Arg(10)-; D048031:JNK Mitogen-Activated Protein Kinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9
Angiogenesis; IL-4; Keratinocyte; Kinin B1 receptor; Kinins; VEGF