论文部分内容阅读
目的:研究新型的特异性类胰蛋白酶抑制剂双苯甲脒,对肥大细胞稳定性的影响。方法:扁桃体组织经酶消化后,将细胞成分用全HEPES缓冲盐溶液(HBSS)重新悬浮。肥大细胞 激发和抑制剂作用的试验在试管中37℃条件下完成。类胰蛋白酶水平用ELISA法测定。结果:同时加入扁桃体细胞悬液中的双苯甲脒,可以剂量依赖的方式抑制抗IgE诱导的类胰蛋白酶释放。仅1 mg/L(1.54μmol/L)的双苯甲脒,即可抑制肥大细胞释放类胰蛋白酶达52％,10 mg/L则能抑制76％的释放。将预培养时间延长30 min,对双苯甲脒的抑制作用无明显影响。在培养到45 min时,双苯甲脒可抑制高达47％的基础类胰蛋白酶释放。与细胞培养15 min后,抗IgE抗体(1g/L)或钙离子导入剂(CI,1 μmol/L)引起的类胰蛋白酶释放的量,分别为基础分泌量的3.2和2.6倍。但是,当细胞与全HBSS预培养超过10 min后,肥大细胞对抗IgE抗体或CI的反应性明显降低。结论:双苯甲脒能够抑制人类扁桃体肥大细胞的IgE依赖性类胰蛋白酶释放。因此,有望开发成为一种新型的肥大细胞稳定剂。
OBJECTIVE: To study the effect of bisbenzamidine, a novel and specific tryptase inhibitor, on the stability of mast cells. METHODS: Tonsil tissue was resuspended with HEPES-buffered saline (HBSS) after enzymatic digestion. Mast cell activation and inhibitor effects were tested in a test tube at 37 ° C. Tryptase levels were measured by ELISA. Results: Simultaneous addition of bis-benzamidine in tonsil cell suspension inhibited anti-IgE-induced tryptase release in a dose-dependent manner. Only 1 mg / L (1.54μmol / L) of bisbenzamidine inhibited the release of tryptase by mast cells up to 52%, while 10 mg / L inhibited 76% of the release. Prolonging the preincubation time for 30 min had no significant effect on the inhibitory effect of bisbenzamidine. Bim benzamidine inhibited up to 47% of basal tryptase release at 45 min. The amount of tryptase released by anti-IgE antibody (1 g / L) or calcium ionomer (CI, 1 μmol / L) at 15 min after cell culture was 3.2 and 2.6 times of the basal secretion, respectively. However, when cells were preincubated with whole HBSS for more than 10 min, the reactivity of mast cells against IgE antibodies or CI was significantly reduced. CONCLUSIONS: Debenzamidine inhibits IgE-dependent tryptase release in human tonsillar mast cells. Therefore, it is expected to develop into a new type of mast cell stabilizer.