›› 2011, Vol. 31 ›› Issue (12): 1667-.doi: 10.3969/j.issn.1674-8115.2011.12.002

• Original article (Basic research) • Previous Articles     Next Articles

Role of ATP release in cyclic pressure-induced mucin excretion in airway epithelial cells

SU Hui-hui, ZHOU Xiang-dong   

  1. Department of Respiratory Medicine, Second Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400010, China
  • Online:2011-12-28 Published:2012-01-04
  • Supported by:

    National Natural Science Foundation of China, 31171346；Cooperation Project of China and Russia of National Natural Science Foundation of China, 81011120108

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Abstract:

Objective To investigate the role of adenosine triphosphate (ATP) release and intra-cellular calcium ion (Ca2+) on cyclic pressure-induced mucin (MUC) excretion in airway epithelial cells. Methods Human bronchial epithelial cell line 16HBE cultured in vitro were stimulated mechanically through tilting the culture dish, and the shear stress and compressive stress were provided by liquid surface tension, atmospheric pressure and liquid gravity. The airway epithelial cells were divided into control group, tilt group, tilt+Ca2+ chelant BAPTA-AM group, tilt+Ca2+ chelant EGTA group and tilt+P2Y receptor blocker RB-2 group. Cell survival rate was measured by MTT assay, expression of MUC5AC mRNA was detected by RT-PCR, expression of MUC5AC protein in culture supernatant was determined by ELISA, ATP release was examined by high performance liquid chromatography (HPLC), and intra-cellular Ca2+ concentration was observed by inverted fluorescence microscopy. Results Compared with control group, the expression of MUC5AC mRNA and protein in 16HBE cells and ATP content in the culture supernatant in tilt group were significantly higher (P<0.05). The expression of MUC5AC protein and ATP content in tilt+BAPTA-AM group and tilt+RB-2 group were significantly lower than those in tilt group (P<0.05). The expression of MUC5AC mRNA in tilt+EGTA group and tilt+RB-2 group was not significantly different from that in tilt group (P>0.05). Conclusion Cyclic mechanical stimulation induces ATP release and MUC5AC excretion in airway epithelial cells in a Ca2+ dependent manner.

Key words: adenosine triphosphate, calcium ion, mucins, cyclic pressure