Mechanisms of asthma and allergic inflammationCysteinyl leukotrienes synergize with growth factors to induce proliferation of human bronchial fibroblasts
Section snippets
Cell cultures
Primary human bronchial fibroblasts were grown from bronchial biopsy specimens obtained from healthy subjects, as previously reported.16 The detailed methods are described in the Methods section in the Online Repository at www.jacionline.org.
Reagents
LTD4, LTC4, and MK-571 were from Cayman Chemical (Ann Arbor, Mich). Montelukast sodium was a kind gift from Dr Jilly Evans (Merck, West Point, Pa). Other reagents are described in the Methods section in the Online Repository at www.jacionline.org.
Mitogenesis analyses
Bronchial
LTD4 synergizes with EGF to induce mitogenesis
To assess whether cys-LTs have mitogenic activity, we analyzed the incorporation of tritiated thymidine into primary cultures of bronchial fibroblasts. LTC4 or LTD4 alone had little mitogenic effect. However, when coincubated with EGF, both LTC4 and LTD4 dose-dependently augmented EGF-induced mitogenesis. Thus 0.5 μmol/L of LTD4 or LTC4 increased mitogenesis by 408% ± 93% (P = .002) or 421% ± 97% (P = .002), respectively, compared with EGF (1 ng/mL) alone. The dose-response curves show very
Discussion
Airway remodeling is an important component of chronic asthma involving proliferation and activation of airway fibroblasts.20 Previous studies have shown that LTC4 and LTD4 stimulate mitogenesis of human skin fibroblasts in vitro.21 Furthermore, targeted deletion of the CysLT1R gene unexpectedly augmented bleomycin-induced lung fibrosis and alveolar septal thickening,22 whereas deletion of CysLT2R prevented these effects.23 These data suggest that CysLT1R and CysLT2R have distinct functions
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Cited by (41)
Lipid mediator Leukotriene D<inf>4</inf>-induces airway epithelial cells proliferation through EGFR/ERK1/2 pathway
2018, Prostaglandins and Other Lipid MediatorsCitation Excerpt :Studies have suggested that GPCRs such as CysLT receptors require growth factor receptors and their tyrosine kinase activity to induce mitogenic effects [9,18]. The CysLT receptors have broader capacity to synergize with other receptor tyrosine kinases (RTKs) and not necessarily transactivate EGFR [10]. We also found that LTD4 caused EGFR phosphorylation concentration-dependently in SAECs and A549 cells which was increased by EGF, though non-significantly, suggesting that the effect of two stimulants could be mediated through EGFR activation (Fig. 4A & B and D & E, respectively), which leads to increased ERK1/2 phosphorylation and activation.
Bradykinin-induced asthmatic fibroblast/myofibroblast activities via bradykinin B<inf>2</inf> receptor and different MAPK pathways
2013, European Journal of PharmacologyCitation Excerpt :In some experiments, cells were pre-incubated with bradykinin B2 receptor antagonist HOE140 (1 μM; 15 min; Sigma-Aldrich) before exposure to bradykinin. The signaling pathways involved in bradykinin-induced HABFb and HNBFb functions were investigated using the specific inhibitors (1 h pretreatment): U0126 (1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene) at 10 µM (for ERK1/2; Sigma-Aldrich) (Davies et al., 2000; Kuang et al., 2007), SB203580 (4-[4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-1H-imidazol-5-yl]pyridine) at 10 µM (for p38 MAPK; Sigma-Aldrich) (Davies et al., 2000; Han et al., 2009), AG1478 (N-(3-chlorophenyl)-6,7-dimethoxy-4-quinazolinanine) at 1 µM (for EGF receptor; Sigma-Aldrich) (Puri et al., 2008; Tsukagoshi et al., 2002), and GM6001 (N-[(2R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl]-l-tryptofan methylamine) at 20 µM (for metalloproteinases; Inalco S.p.A, Milan, Italy) (Devel et al., 2012; Yoshisue et al., 2007). ERK1/2 and p38 activation were evaluated in fibroblasts stimulated with bradykinin (10−11 or 10−6 M) for 5 min.
Effects of montelukast on subepithelial/peribronchial fibrosis in a murine model of ovalbumin induced chronic asthma
2013, International ImmunopharmacologyCitation Excerpt :Montelukast inhibits the action of leukotriene D4 on the cysteinyl leukotriene receptor (CysLTR)1 in the lung, which reduces bronchoconstriction and airway inflammation [8,9]. Previous studies have demonstrated that montelukast attenuates the airway remodeling induced by several allergens by modulating the expression of Th2 cytokines and growth factors [10–12]. In addition to these features, several researchers have demonstrated that montelukast exerts beneficial effects against experimentally induced toxicities.
Cysteinyl leukotrienes regulate TGF-Β<inf>1</inf> and collagen production by bronchial fibroblasts obtained from asthmatic subjects
2012, Prostaglandins Leukotrienes and Essential Fatty AcidsCitation Excerpt :We showed an elevated expression of CysLT1R by bronchial fibroblasts from asthmatics compared to cells from non-asthmatics. Yoshisue et al. have reported that primary bronchial human fibroblasts from healthy controls express a weak signal for CysLT1R [32]. This observation is challenged by other studies showing that these cells express this receptor, express constitutively 5-lipoxygenase and have the enzymatic machinery to produce CysLTs [33].
Supported by Asthma UK.
Disclosure of potential conflict of interest: H. Yoshisue and D. E. Davies have received grant support from Asthma UK. A. P. Sampson has consultant arrangements with Merck Sharp & Dohme; has received grant support from Merck & Co, Ono Pharma UK Ltd, and AstraZeneca; and has received honoraria from Merck & Co. The rest of the authors have declared that they have no conflict of interest.