中文摘要：

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-CO₂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α₁-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。

英文摘要：

AIM: To investigate the actions of protease inhibitors on the enzymatic activities of tryptase and chymase in similar experimental systems. METHODS: Human lung tryptase and human skin chymase were purified by a similar procedure involving high salt extraction of tryptase, heparin agarose affinity chromatography, and S-200 Sephacryl gel filtration chromatography. Actions of protease inhibitors on tryptase and chymase activities were examined by enzyme assays. RESULTS: The specific activities of tryptase and chymase were 2.1 kU/g protein and 4.9 kU/g protein, respectively. Both preparations showed a single diffuse band on SDS-PAGE. Among non-native protease inhibitors, N-（1-hydroxy-2-naphthoyl）-L-arginyl-L-prolinamide hydrochloride （HNAP）, leupeptin, antipain, benzamidine, and protamine inhibited more than 90% enzymatic activity of tryptase, whereas soy bean trypsin inhibitor （SBTI）, Z-Ile-Glu-Pro-Phe-CO₂Me （ZIGPPM） and chymostatin inhibited more than 95% enzymatic activity of chymase. Native protease inhibitos α₁-antitrypsin and secretory leukocyte protease inhibitor （SLPI） inhibited more than 90% enzymatic activity of chymase, but lactoferrin appeared to enhance chymase enzymatic activity. All the 3 inhibitors had weak inhibitory actions on tryptase. CONCLUSION: The protease inhibitors tested had relatively good selectivity to either tryptase or chymase.