Effect of nitric oxide donation on mucin production in vitro.

Otitis media with effusion (OME) is characterized by the accumulation of a viscous fluid rich in mucins in the middle ear cleft. There is increasing evidence that this fluid is the result of an inflammatory reaction and that nitric oxide (NO) is an important mediator in this reaction. The goblet cell line HT29-MTX produces principally MUC5AC, an important mucin in middle ear effusions, and thus is a good model for the study of mucus-secreting epithelia. Confluent cell cultures were trypsinized, subcultured and incubated with isosorbide dinitrate (ISDN), a NO donor, for 0.5, 1 and 2 h at a concentration of 1 mm and in concentrations of 0.01, 0.1, 0.5, 1 and 2 mm for 1 h. Experiments were performed four times. Mucin production was detected by a slot blot ELISA assay, using a monoclonal mouse antibody to human MUC5AC mucin. Statistical significance was tested using a one-way analysis of variance. NO donation by ISDN caused a consistent rise in mucin production above control. Maximal mucin production of 35% above control occurred at 1 h with 1 mm ISDN. Mucin production increased from 12% above control with 0.1 mm ISDN dinitrate to 45% above baseline with 2 mm ISDN. NO donation by ISDN results in an increase in mucus production, which is both dose and time related. This adds further evidence to an inflammatory model for mucus secretion in OME.