摘要
AIM: To investigate the tryptase and histamine release ability of human colon mast cells upon IgE dependent or independent activation and the potential mechanisms.METHODS: Enzymatically dispersed cells from human colons were challenged with anti-IgE or calcium ionophore A23187, and the cell supernatants after challenge were collected. Both concentration dependent and time course studies with anti-IgE or calcium ionophore A23187 were performed. Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibre-based fiuorometric assay.RESULTS:Both anti-IgE and calcium ionophore were able to induce dose dependent release of histamine from colon mast cells with up to approximately 60% and 25% net histamine release being achieved with 1μg/mL calcium ionophore and 10μg/mL anti-IgE, respectively. Dose dependent release of tryptase was also observed with up to approximately 19ng/mL and 21ng/mL release of tryptase being achieved with 10μg/mL anti-IgE and 1μg/mL calcium ionophore, respectively. Time course study revealed that both tryptase and histamine release from colon mast cells stimulated by anti-IgE initiated within 10 sec and reached their maximum release at 6 min following challenge. Pretreatment of cells with metabolic inhibitors abolished the actions of anti-IgE as well as calcium ionophore. Tryptase and histamine release, particularly that induced by calcium ionophore was inhibited by pretreatment of cells with pertussis toxin.CONCLUSION: Both anti-IgE and calcium ionophore are able to induce significant release of tryptase and histamine from colon mast cells, indicating that this cell type is likely to contribute to the pathogenesis of colitis and other mast cell associated intestinal diseases.
AIM:To investigate the tryptase and histamine release ability of human colon mast cells upon IgE dependent or independent activation and the potential mechanisms. METHODS:Enzymatically dispersed cells from human colons were challenged with anti-IgE or calcium ionophore A23187,and the cell supernatants after challenge were collected.Both concentration dependent and time course studies with anti-IgE or calcium ionophore A23187 were performed.Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibre-based fiuorometric assay. RESULTS:Both anti-IgE and calcium ionophore were able to induce dose dependent release of histamine from colon mast cells with up to approximately 60% and 25% net histamine release being achieved with 1 μg/mL calcium ionophore and 10μg/mL anti-IgE,respectively.Dose dependent release of tryptase was also observed with up to approximately 19 ng/mL and 21 ng/mL release of tryptase being achieved with 10μg/mL anti-IgE and 1μg/ mL calcium ionophore,respectively.Time course study revealed that both tryptase and histamine release from colon mast cells stimulated by anti-IgE initiated within 10 sec and reached their maximum release at 6 rain following challenge.Pretreatment of cells with metabolic inhibitors abolished the actions of anti-IgE as well as calcium ionophore.Tryptase and histamine release,particularly that induced by calcium ionophore was inhibited by pretreatment of cells with pertussis toxin. CONCLUSION:Both anti-IgE and calcium ionophore are able to induce significant release of tryptase and histamine from colon mast cells,indicating that this cell type is likely to contribute to the pathogenesis of colitis and other mast cell associated intestinal diseases.
基金
Supported by the National Natural Science Foundation of China,No.30140023,and the Li Ka Shing Foundation,Hong Kong,China,No.C0200001
