Inflammatory conditions increase expression of protease‐activated receptor‐2 by human airway smooth muscle cells in culture

Abstract

The protease-activated receptor-2 (PAR-2) has been implicated in airway inflammation and bronchial hyperresponsiveness. We wondered whether inflammatory conditions may upregulate PAR-2 expression by the human airway smooth muscle. To do so, we treated human airway smooth muscle cells (HASMC) in primary culture with interleukin-1beta (IL-1beta), a pro-inflammatory and asthma-associated cytokine. Cells were starved for 24 h and incubated with or without IL-1beta. Online fluorescent polymerase chain reaction after reverse transcription quantified PAR-2 mRNA, and Western blotting measured PAR-2 protein expression. PAR-2 was constitutively expressed by HASMC in primary culture, and IL-1beta (10 U/mL) time dependently elevated PAR-2 mRNA with a maximum of 4.7-fold after 1.5 h (P < 0.01), and PAR-2 protein expression with a maximum of 1.5-fold after 24 h (P < 0.01). The concentration dependence of the IL-1beta effect (0.1-30 U/mL) confirmed a maximal increase of PAR-2 expression at 10 U/mL. Our study clearly shows that IL-1beta upregulates PAR-2 mRNA and protein expression by HASMC in culture. This increased expression of PAR-2 in inflammatory conditions may have functional consequences in the bronchial dysfunction of asthmatic airways.