目的:在同一实验系统中研究蛋白酶抑制剂对人类肥大细胞类胰蛋白酶和类糜蛋白酶催化活性的调节作用。方法:应用高盐提取、肝素凝胶亲和层析和S-200凝胶过滤层析的方法纯化人肺类胰蛋白酶和人皮肤类糜蛋白酶,蛋白酶抑制剂对类胰蛋白酶和类糜蛋白酶催化活性的调节作用则由酶-底物反应实验来测定。结果:纯化后的类胰蛋白酶和类糜蛋白酶的比活分别为2.1U/mg蛋白质和4.9U/mg蛋白质。两种纯化产物在SDS-PAGE上均显示为单一条带。非内源性蛋白酶抑制剂N-(1-羟基-2-萘基)-L-精氨酰-L-脯氨酰胺-盐酸、亮肽素、antipain、苯甲脒、鱼精蛋白可抑制90％的类胰蛋白酶催化活性,而大豆胰蛋白酶抑制剂、Z-异亮氨酸-谷氨酸-脯氨酸-苯丙氨酸-CO_2甲基、抑凝乳蛋白酶素则抑制类糜蛋白酶催化活性的95％,内源性蛋白酶抑制剂α_1-抗胰蛋白酶和分泌性白细胞蛋白酶抑制剂抑制90％以上的类糜蛋白酶催化活性,但乳铁蛋白却表现出具有增加类糜蛋白酶催化活性的作用,这三种内源性蛋白酶抑制剂均表现出较弱的抑制类胰蛋白酶的作用。结论:本实验所采用的蛋白酶抑制剂具有相对好的类胰蛋白酶或类糜蛋白酶选择性,表明它们可被应用于抑制剂药物的开发过程中。 OBJECTIVE: To study the regulatory effect of protease inhibitors on the catalytic activity of human mast cell tryptase and chymase in the same experimental system. Methods: Human lung trypsin and human skin chymase were purified by high salt extraction, heparin gel affinity chromatography and S-200 gel filtration chromatography. Protease inhibitors were used to catalyze tryptase and chymase Activity regulation by the enzyme - substrate reaction to determine the experiment. Results: The specific activities of purified tryptase and chymase were 2.1 U / mg protein and 4.9 U / mg protein, respectively. Both purified products showed a single band on SDS-PAGE. Non-endogenous protease inhibitors N- (1-hydroxy-2-naphthyl) -L-arginyl-L-prolinamide hydrochloride, leupeptin, antipain, benzamidine, protamine can inhibit 90 % Of the tryptase catalytic activity, and soybean trypsin inhibitor, Z-isoleucine - glutamic acid - proline - phenylalanine - CO2 methyl, chymostatin inhibited chymase catalytic 95% of the activity, the endogenous protease inhibitors α 1-antitrypsin and secreted leukocyte protease inhibitors inhibit more than 90% of the chymotrypsin catalytic activity, but lactoferrin has been shown to increase the chymotrypsin catalytic activity , All three endogenous protease inhibitors showed weaker inhibition of tryptase. CONCLUSIONS: The protease inhibitors used in this experiment have relatively good tryptase or chymase selectivity, suggesting that they may be used in the development of inhibitor drugs.