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Regulation of PMA-induced MUC5AC expression by heparin in human bronchial epithelial cells

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Abstract

Mucus hypersecretion is a major pathophysiologic feature in chronic inflammatory airway diseases. Oxidative stress plays a pivotal role in this process. Recent studies have found that heparin has antioxidant effects which can reduce free radical damage. Here, we hypothesized that heparin has some influence on the expression of mucin 5AC (MUC5AC) induced by phorbol myristate acetate (PMA) in a bronchial epithelial cell line (HBE16), also we have investigated the potential mechanism involved in the process. We found that ROS, the mRNA of Duox1, EGFR and MUC5AC, as well as the protein levels of Duox1, p-EGFR, EGFR, and MUC5AC in the PMA group were significantly increased when compared with the control group (all P < 0.01). After pretreatment with heparin however, there was a significant decrease in ROS levels, the mRNA of Duox1, EGFR, and MUC5AC, and the protein levels of Duox1, p-EGFR, EGFR, and MUC5AC, when compared with the PMA group (all P < 0.01). MUC5AC protein in the supernatant was inhibited in a dose-dependent manner by heparin. Pretreatment with DMTU resulted in a significant decrease in ROS content, the mRNA of Duox1, EGFR, and MUC5AC as well as the protein levels of Duox1, p-EGFR, EGFR, and MUC5AC when compared with the PMA group (all P < 0.01). When cells were pretreated with both heparin and DMTU, there was a further reduction in ROS content, the mRNA of Duox1, EGFR, and MUC5AC as well as the protein levels of Duox1, p-EGFR, EGFR, and MUC5AC, when compared with either the PMA group, heparin group, or DMTU group (all P < 0.01). Our results show that PMA can induce MUC5AC expression by activation of the Duox1-ROS-TACE-TGF-α-EGFR signaling pathway. Heparin can decrease the level of Duox1, ROS production and block the PMA-induced activation of EGFR, thus inhibiting the overexpression of mucin MUC5AC in a dose-dependent manner. In addition to reducing ROS production, heparin may also inhibit the expression of MUC5AC through other signal mechanisms.

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Abbreviations

DMTU:

Dimethylthiourea

Duox1:

Dual oxidase 1

EGFR:

Epidermal growth factor receptor

HNE:

Human neutrophil elastase

MUC5AC:

Mucin 5AC

PMA:

Phorbol myristate acetate

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Acknowledgments

This study was supported by grant from the National Nature Science Foundation of China (No. 81070031), and China–Russia Cooperation Research Foundation (No. 81011120108). There are no conflict of interest to declare.

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Authors and Affiliations

  1. Department of Respiratory Medicine, The Second Affiliated Hospital, Chongqing Medical University, No.74, Linjiang Road, Yuzhong District, Chongqing, 400010, China

    Qi Li, Rui Xia Lei & Xiang Dong Zhou

  2. Far Eastern Scientific Center of Physiology and Pathology of Respiration, Blagoveschensk, 675000, Russia

    Victor P. Kolosov & Juliy M. Perelman

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  1. Qi Li
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  2. Rui Xia Lei
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  3. Xiang Dong Zhou
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Correspondence to Xiang Dong Zhou.

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Li, Q., Lei, R.X., Zhou, X.D. et al. Regulation of PMA-induced MUC5AC expression by heparin in human bronchial epithelial cells. Mol Cell Biochem 360, 383–391 (2012). https://doi.org/10.1007/s11010-011-1078-9

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  • DOI: https://doi.org/10.1007/s11010-011-1078-9

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