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Chronic mechanical stress induces mucin5AC expression in human bronchial epithelial cells through ERK dependent pathways

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Abstract

Mucus hypersecretion is a common pathological change in chronic inflammatory diseases of the airway. These conditions are usually accompanied by chronic mechanical stress due to airway constriction. Our objective was to study the molecular mechanisms and physical effects of chronic mechanical stress on mucin 5AC (MUC5AC) expression in airway epithelial cells. We exposed normal human bronchial epithelial (NHBE) cells cultured at an air–liquid interface to different degrees of chronic compressive mechanical stress (10, 20, 30 cmH2O) for 7 days(1 h per day). MUC5AC protein content was detected by enzyme-linked immunosorbent assay (ELISA). MUC5AC mRNA expression was detected by reverse transcription PCR (RT-PCR) and real-time PCR. The effects of chronic mechanical stress on phosphorylated ERK1/2 (p-ERK1/2), phosphorylated JNK (p-JNK), phosphorylated P38 (p-P38), and phosphorylation of FAK at Tyr397 (p-FAK-Y397), were assessed by Western blot. We also assessed the impact of, an EGFR kinase inhibitor (AG1478), an ERK kinase inhibitor (PD-98059), and short interfering RNA (siRNA) targeted to FAK. We found that transcriptional and protein expression levels of MUC5AC were elevated significantly in the 30 cmH2O compressive stress group. p-ERK1/2 increased significantly in response to compressive stress and PD-98059 could attenuated stress-induced MUC5AC expression. p-FAK-Y397 increased significantly in response to compressive stress and FAK siRNA attenuated stress-induced ERK activation strongly. AG1478 attenuated stress-induced ERK activation and MUC5AC expression significantly, but incompletely. Combination of FAK siRNA and AG1478 led to complete attenuation of ERK activation and MUC5AC expression. These results suggest that chronic mechanical stress can enhance MUC5AC expression in human bronchial epithelial cells through the ERK signal transduction pathway. Both FAK and EGFR mediate the mitogenic response induced by mechanical stress in human bronchial epithelial cells through an ERK signaling cascade.

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Acknowledgments

This work was supported by a grant from the National Nature Science Foundation of China (No.81070031), and the China-Russia Cooperation Research Foundation (No.81011120108).

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Correspondence to Xiang Dong Zhou.

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Li, N., Li, Q., Zhou, X.D. et al. Chronic mechanical stress induces mucin5AC expression in human bronchial epithelial cells through ERK dependent pathways. Mol Biol Rep 39, 1019–1028 (2012). https://doi.org/10.1007/s11033-011-0827-x

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  • DOI: https://doi.org/10.1007/s11033-011-0827-x

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