摘要
AIM:To investigate the ability of agonists of PAR-2 to stimulate release of tryptase and histamine from human colon mast cells and the potential mechanisms.METHODS:Enzymatically dispersed cells from human colons were challenged with tc-LIGRLO, tc-OLRGIL, SLIGKV,VKGILS, trypsin, anti-IgE or calcium ionophore A23187,and the cell supematants after challenge were collected. Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibrebased fluorometric assay.RESULTS: Both PAR-2 agonists tc-LIGRLO-NH2 and SLIGKVNH2 were able to induce dose dependent release of tryptase and histamine from colon mast cells. More than 2.5 fold increase in both tryptase and histamine release was provoked by 100μmol/mL tc-LIGRLO-NH2, in comparison with only 2.0 fold increase being stimulated by SLIGKV-NH2，The reverse peptides tc-OLRGIL-NH2 and VKGILS-NH2 at the concentrations tested had no effect on the release of these two mediators.The maximum tryptase release elicited by tc-LIGRLO-NH2 was similar to that induced by anti-IgE(10μg/mL) or calcium ionophore (1μg/mL), though the latter was a more potent stimulus for histamine release.Both histamine and tryptase release in response to tc-LIGRLONH2 were completed within 3 rain. Trypsin at concentrations from 1.0 to 100μg/mL was capable of provoking a dose dependent release of tryptase as well as histamine with a maximum of 16ng/mL tryptase and 14ng/mL histamine release being achieved. An approximately 80% and 70% inhibition of trypsin induced release of tryptase and histamine were observed with SBTI, respectively. Pretreatment of cells with metabolic inhibitors or pertussis toxin abolished the actions of tc-LIGRLO-NH2, SLIGKV-NH2 and trypsin.CONCLUSION: The agonists of PAR-2 and trypsin are potent secretagogues of human colon mast cells, which are likely to contribute to the development of inflammatory disorders in human gut.
AIM:To investigate the ability of agonists of PAR-2 to stimulate release of tryptase and histamine from human colon mast cells and the potential mechanisms. METHODS:Enzymatically dispersed cells from human colons were challenged with tc-LIGRLO,tc-OLRGIL,SLIGKV, VKGILS,trypsin,anti-IgE or calcium ionophore A23187,and the cell supernatants after challenge were collected.Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibre- based fluorometric assay. RESULTS:Both PAR-2 agonists tc-LIGRLO-NH_2 and SLIGKV- NH_2 were able to induce dose dependent release of tryptase and histamine from colon mast cells.More than 2.5 fold increase in both tryptase and histamine release was provoked by 100μmol/mL tc-LIGRLO-NH_2,in comparison with only 2.0 fold increase being stimulated by SLIGKV-NH_2.The reverse peptides tc-OLRGIL-NH_2 and VKGILS-NH_2 at the concentrations tested had no effect on the release of these two mediators.The maximum tryptase release elicited by tc-LIGRLO-NH_2 was similar to that induced by anti-IgE (10μg/mL)or calcium ionophore(1μg/mL),though the latter was a more potent stimulus for histamine release.Both histamine and tryptase release in response to tc-LIGRLO- NH_2 were completed within 3 min.Trypsin at concentrations from 1.0 to 100μg/mL was capable of provoking a dose dependent release of tryptase as well as histamine with a maximum of 16 ng/mL tryptase and 14 ng/mL histamine release being achieved.An approximately 80% and 70% inhibition of trypsin induced release of tryptase and histamine were observed with SBTI,respectively.Pretreatment of cells with metabolic inhibitors or pertussis toxin abolished the actions of tc-LIGRLO-NH_2,SLIGKV-NH_2 and trypsin. CONCLUSION:The agonists of PAR-2 and trypsin are potent secretagogues of human colon mast cells,which are likely to contribute to the development of inflammatory disorders in human gut.
基金
Supported by the National Natural Science Foundation of China,No.30140023,and the Li Ka Shing Foundation,Hong Kong,China,No.C0200001
