title: Protease activated receptor-2 expression and function in asthmatic bronchial smooth muscle.
abstract: Asthmatic bronchial smooth muscle (BSM) is characterized by structural remodeling associated with mast cell infiltration displaying features of chronic degranulation. Mast cell-derived tryptase can activate protease activated receptor type-2 (PAR-2) of BSM cells. The aims of the present study were (i) to evaluate the expression of PAR-2 in both asthmatic and non asthmatic BSM cells and, (ii) to analyze the effect of prolonged stimulation of PAR-2 in asthmatic BSM cells on cell signaling and proliferation. BSM cells were obtained from both 33 control subjects and 22 asthmatic patients. PAR-2 expression was assessed by flow cytometry, western blot and quantitative RT-PCR. Calcium response, transduction pathways and proliferation were evaluated before and following PAR-2 stimulation by SLIGKV-NH2 or trypsin for 1 to 3 days. Asthmatic BSM cells expressed higher basal levels of functional PAR-2 compared to controls in terms of mRNA, protein expression and calcium response. When PAR-2 expression was increased by means of lentivirus in control BSM cells to a level similar to that of asthmatic cells, PAR-2-induced calcium response was then similar in both types of cell. However, repeated PAR-2 stimulations increased the proliferation of asthmatic BSM cells but not that of control BSM cells even following lentiviral over-expression of PAR-2. Such an increased proliferation was related to an increased phosphorylation of ERK in asthmatic BSM cells. In conclusion, we have demonstrated that asthmatic BSM cells express increased baseline levels of functional PAR-2. This higher basal level of PAR-2 accounts for the increased calcium response to PAR-2 stimulation, whereas the increased proliferation to repeated PAR-2 stimulation is related to increased ERK phosphorylation.
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Allard
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Imane
authors.initials: I
authors.lastname: Bara
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Gilbert
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Carvalho
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Trian
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France ; CHU de Bordeaux, Service d'Exploration Fonctionnelle Respiratoire, Pessac, France.
authors.forename: Annaig
authors.initials: A
authors.lastname: Ozier
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Gillibert-Duplantier
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Olga
authors.initials: O
authors.lastname: Ousova
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Elise
authors.initials: E
authors.lastname: Maurat
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France ; CHU de Bordeaux, Service d'Exploration Fonctionnelle Respiratoire, Pessac, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Thumerel
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Quignard
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France ; CHU de Bordeaux, Service d'Exploration Fonctionnelle Respiratoire, Pessac, France.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Girodet
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France ; CHU de Bordeaux, Service d'Exploration Fonctionnelle Respiratoire, Pessac, France.
authors.forename: Roger
authors.initials: R
authors.lastname: Marthan
authors.affiliation: Univ- Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Département de Pharmacologie, Bordeaux, France ; INSERM, Centre de Recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France ; CHU de Bordeaux, Service d'Exploration Fonctionnelle Respiratoire, Pessac, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Berger
mesh_terms: D000328:Adult; D000368:Aged; D001249:Asthma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D001980:Bronchi / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002118:Calcium / Q000378:metabolism; D016022:Case-Control Studies; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation; D002478:Cells, Cultured; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D009842:Oligopeptides / Q000494:pharmacology; D010766:Phosphorylation; D044464:Receptor, PAR-2 / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014357:Trypsin / Q000494:pharmacology; D053802:Tryptases / Q000235:genetics / Q000378:metabolism
chemical_list: D009842:Oligopeptides; D044464:Receptor, PAR-2; D048049:Extracellular Signal-Regulated MAP Kinases; D014357:Trypsin; D053802:Tryptases; D002118:Calcium