The B1-agonist [des-Arg10 ]-kallidin Activates Transcription Factor NF-B and Induces Homologous Upregulation of the Bradykinin B1-receptor in Cultured Human Lung Fibroblasts

Joost Schanstra (1) , E Bataillé (2, 3) , Maria E. Marin Castaño (1) , Yannick Barascud (1) , Christophe Hirtz (4) , João B. Pesquero (5) , Christiane Pecher (6) , Francis Gauthier (2) , Jean-Pierre Girolami (6) , Jean-Loup Bascands (6)

1 INSERM - Institut National de la Santé et de la Recherche Médicale
2 CNRS - Centre National de la Recherche Scientifique
3 Université Francois Rabelais [Tours]
4 UM - Université de Montpellier
5 MDC - Max Delbrück Center for Molecular Medicine [Berlin]
6 I2MR - Institut de médecine moléculaire de Rangueil

The bradykinin B1-receptor is strongly upregulated under chronic inflammatory conditions. However, the mechanism and reason are not known. Because a better understanding of the mechanism of the upregulation will help in understanding its potential importance in inflammation, we have studied the molecular mechanism of B1-receptor upregulation in cultured human lung fibroblasts (IMR 90) in response to IL-1beta and the B1-agonist [des-Arg10]-kallidin. We show that treatment of human IMR 90 cells by IL-1beta stimulates the expression of both B1-receptor mRNA and protein. The latter was studied by Western blot analysis using antipeptide antibodies directed against the COOH-terminal part of the human B1-receptor. We furthermore report the novel observation that the B1-receptor is upregulated by its own agonist which was completely blocked by the specific B1-antagonist [des-Arg10-Leu9]-kallidin, indicating an upregulation entirely mediated through cell surface B1-receptors. The increased population of B1-receptors was functionally coupled as exemplified by an enhancement of the B1-agonist induced increase in free cytosolic calcium. Upregulation by the B1-agonist was blocked by a specific protein kinase C inhibitor. B1-agonist-induced upregulation was correlated to the induction of transcription factor nuclear factor kappaB (NF-kappaB) which efficiently bound to the NF-kappaB-like sequence located in the promoter region of the human B1-receptor gene. This correlation was further confirmed by reporter gene assays which showed that this NF-kappaB-like sequence, in the B1-receptor promoter context, could contribute to IL-1beta and DLBK-induced B1-receptor transcription activation, and by the effect of NF-kappaB inhibitor pyrrolidinedithiocarbamate which diminished both B1-receptor upregulation and NF-kappaB activation. NF-kappaB is now recognized as a key inflammatory mediator which is activated by the B1-agonist but which is also involved in B1-receptor upregulation.

Mots clés: NF-kB; gene expression; inflammation; B1-receptor

Domaines: Biochimie, Biologie Moléculaire; Interactions cellulaires [q-bio.CB]

Joost Schanstra, E Bataillé, Maria E. Marin Castaño, Yannick Barascud, Christophe Hirtz, et al.. The B1-agonist [des-Arg10 ]-kallidin Activates Transcription Factor NF-B and Induces Homologous Upregulation of the Bradykinin B1-receptor in Cultured Human Lung Fibroblasts. Journal of Clinical Investigation, 1998, 101 (10), pp.2080-2091. ⟨10.1172/JCI1359⟩. ⟨hal-02502902⟩