鱼精蛋白对人结肠肥大细胞IgE依赖性与非依赖性类胰蛋白酶释放的调节

The Ability of Protamine to Modulate Tryptase Release from Human Colon Mast Cells on IgE Dependent and Non-IgE Dependent Stimulation

目的 :探讨鱼精蛋白对人结肠肥大细胞IgE依赖性与非依赖性类胰蛋白酶释放的调节。方法 :人结肠组织经酶消化后 ,细胞成分用全HBSS重新悬浮。激发过程在LP4 试管中、 37°C条件下完成。用酶联免疫吸附试验方法测定类胰蛋白酶水平。结果 :促分泌剂抗IgE抗体和离子载体钙 (CI)均可明显刺激人结肠肥大细胞类胰蛋白酶的释放。鱼精蛋白浓度在<10 μg mL时可以剂量依赖性的方式抑制抗IgE抗体和CI诱导的类胰蛋白酶释放 ,但是 ,浓度为 10 0 μg mL时 ,作用则相反。在 37°C条件下同结肠细胞预培养 2 0min ,1μg mL的鱼精蛋白可明显抑制抗IgE抗体和CI诱导的类胰蛋白酶释放 ,而 10 μg mL的鱼精蛋白则起相反的作用。结论 :小剂量鱼精蛋白可以抑制肥大细胞IgE依赖性与非依赖性类胰蛋白酶的释放 。

Objective:To investigate the ability of protamine to modulate tryptase release from human colon mast cells on IgE dependent and non_IgE dependent stimulation.Methods:Human mast cells were dispersed from colon tissue with collagenase and hyaluronidase, and were challenged with stimulus for 15 minutes at 37°C.Tryptase concentrations were measured with a sandwich ELISA procedure. Results:Anti_IgE(10μg/mL)and calcium ionophore(CI)(1μg/mL)were able to provoke significant tryptase release from human colon mast cells,but protamine was not.Up to 10μg/mL,protamine was able to inhibit anti_IgE and CI induced tryptase release in a concentration dependent manner with a maximum of 20.9% and 17.1% inhibition,respectively.At the concentration of 100μg/mL,protamine was able to provoke anti_IgE and CI induced tryptase release with 54.7% and 24.9%, respectively.Preincubation of protamine with cells for 20 minutes at 37°C before challenging with anti_IgE or CI,1g/mL of protamine had ability to inhibit tryptase release from colon mast cells,10g/mL of protamine had a reverse action.Conclusion:Protamine at low concentration is able to inhibit anti_IgE and CI induced tryptase release from human colon mast cells, which may indicate a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cell related diseases.