Mechanism of montelukast on autophagy and apoptosis of airway epithelial cells through STAT3-RORγt-IL-17/IL-23 signaling pathway

Abstract

This study investigates the mechanism of montelukast intervention on autophagy and apoptosis of airway epithelial cells. Construction of HBE cell building model, which were intervened by montelukast. The proliferation of human 16HBE cells was detected using MTT method and β-catenin level was detected. The cellular cycle distribution, autophagy and apoptosis were detected using flow cytometry. And expressions of Transcriptional activator 3 (STAT3)-Retinoic acid-associated nuclear orphan receptor (RORγt)-interleukin 17 (IL-17)/interleukin 23 (IL-23) signaling related proteins were measured using Western blot. Montelukast inhibited the proliferation of human 16HBE cells and its inhibition rate and action concentration showed time and dose dependence. The half maximal inhibitory concentrations (IC50) were (12.8±0.67) μmol/L at 24 h, (8.8±0.43) μmol/L at 48 h and (6.6±0.42) μmol/L at 72 h, respectively. Montelukast induced 16HBE cellular cycle to arrest in G2/M phase dose-dependently (5, 10 and 20 μmol/L) (P <0.05) and simultaneously increased apoptosis rate (P < 0.05). 40 μL montelukast had a protective effect on 16HBE cells. In addition, montelukast reduced β-catenin level, which suggested that STAT3-RORγt-IL-17/IL-23 signaling pathway might be inhibited. Meanwhile, montelukast reduced the expressions of STAT3, P-STAT3, RORγt (RORβ), c-myc and survivin and increased protein expressions of GSK-3 (RORα) and Th17, but had no effect on the total RORγt level. Montelukast may effectively promote the apoptosis of 16HBE airway epithelial cells via inhibition of STAT3-RORγt-IL-17/IL-23 signaling.