Xie Hua, He Shao-heng, Cheng Ming-hua, Fu Yi-ling
Allergy & Inflammation Research Institute, Medical College of Shantou University, Shantou 515031, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Nov;20(6):678-81.
To investigate the effect of tryptase inhibitors (TPIs) on histamine release from human colon mast cells.
Human mast cells were prepared by digestion of colon tissue and with collagenase and hyaluronidase, cultured with four kinds of TPIs, leupeptin, protamine, TLCK, and lactoferrin for 15 min at 37 degrees Celsius respectively. A glass fibre-based fluorometry assay was used to detect histamine in mast cell suspension.
200 mmol/L leupeptin and 100 mmol/L protamine were able to stimulate histamine release from colon mast cells, while TLCK and lactoferrin did not. All TPIs inhibited anti-IgE-induced histamine release in a concentration dependent manner, and the inhibitory rates were 48.7%, 36.7%, 40.2% and 34.1%, respectively. However preincubation of TPIs with mast cells for 20 min at 37 degrees Celsius before adding anti-IgE had little effect on anti-IgE induced histamine release. All TPIs were able to inhibit calcium ionophore (CI)-induced histamine release, and the maximum inhibition rate was between 25%-32%. Inhibition on histamine release by leupeptin and TLCK obviously enhanced when colon mast cells were preincubated with them for 20 min before adding CI. However, under the same condition, protamine failed to inhibit histamine release.
We prove for the first time that TPIs inhibit anti-IgE-and CI-induced histamine release from human colon mast cells, suggesting that it is possible to treat inflammatory bowel disease or other mast cell-related diseases by using TPIs.
研究类胰蛋白酶抑制剂(TPIs)对人结肠肥大细胞组胺释放的影响。
通过用胶原酶和透明质酸酶消化结肠组织制备人肥大细胞,分别与四种TPIs(亮抑酶肽、鱼精蛋白、甲苯磺酰-L-赖氨酸氯甲基酮和乳铁蛋白)在37摄氏度下培养15分钟。采用基于玻璃纤维的荧光测定法检测肥大细胞悬液中的组胺。
200 mmol/L亮抑酶肽和100 mmol/L鱼精蛋白能够刺激结肠肥大细胞释放组胺,而甲苯磺酰-L-赖氨酸氯甲基酮和乳铁蛋白则不能。所有TPIs均以浓度依赖性方式抑制抗IgE诱导的组胺释放,抑制率分别为48.7%、36.7%、40.2%和34.1%。然而,在加入抗IgE之前,将TPIs与肥大细胞在37摄氏度下预孵育20分钟,对抗IgE诱导的组胺释放影响不大。所有TPIs均能抑制钙离子载体(CI)诱导的组胺释放,最大抑制率在25%-32%之间。当结肠肥大细胞在加入CI之前与亮抑酶肽和甲苯磺酰-L-赖氨酸氯甲基酮预孵育20分钟时,对组胺释放的抑制作用明显增强。然而,在相同条件下,鱼精蛋白未能抑制组胺释放。
我们首次证明TPIs抑制人结肠肥大细胞抗IgE和CI诱导的组胺释放,提示使用TPIs治疗炎症性肠病或其他肥大细胞相关疾病是可能的。