Barna J B, Kew R R
Department of Pathology State University of New York at Stony Brook 11794-8691, USA.
Inflammation. 1995 Oct;19(5):561-74. doi: 10.1007/BF01539136.
Previous studies have demonstrated that neutrophils possess an active serine protease(s) which may be involved in the process of chemotaxis but the precise identity of this enzyme(s) remains to be determined. In this study fourteen different protease inhibitors were tested over a wide concentration range for their ability to inhibit unstimulated neutrophil movement and chemotaxis to C5a, fMLP and IL-8. Pretreatment of neutrophils with aspartyl or metallo-protease inhibitors had no effect on either chemotaxis or random cell movement. The thiol protease inhibitors E-64 and cystatin, as well as the thiol/serine inhibitors antipain and leupeptin, diminished only C5a-induced chemotaxis. Pretreatment of neutrophils with the serine protease inhibitors PMSF or 3,4-DCI significantly reduced chemotaxis to C5a, fMLP and IL-8. The inhibitor of trypsin-like serine proteases, TLCK, and the neutrophil elastase inhibitor MeO-Suc-AAPV-CMK had no inhibitory effect on cell movement. However, two different inhibitors of chymotrypsin-like serine proteases, TPCK and chymostatin, significantly inhibited movement to any chemoattractant. These results suggest that an active chymotrypsin-like serine protease is essential for neutrophils to respond to chemotactic stimuli.
先前的研究表明，中性粒细胞拥有一种活性丝氨酸蛋白酶，其可能参与趋化作用过程，但这种酶的确切身份仍有待确定。在本研究中，测试了14种不同的蛋白酶抑制剂在较宽浓度范围内抑制未受刺激的中性粒细胞运动以及对C5a、fMLP和IL-8趋化作用的能力。用天冬氨酸蛋白酶或金属蛋白酶抑制剂预处理中性粒细胞，对趋化作用或随机细胞运动均无影响。巯基蛋白酶抑制剂E-64和胱抑素，以及巯基/丝氨酸抑制剂抗蛋白酶和亮抑酶肽，仅减弱C5a诱导的趋化作用。用丝氨酸蛋白酶抑制剂PMSF或3,4-DCI预处理中性粒细胞，可显著降低对C5a、fMLP和IL-8的趋化作用。胰蛋白酶样丝氨酸蛋白酶抑制剂TLCK和中性粒细胞弹性蛋白酶抑制剂MeO-Suc-AAPV-CMK对细胞运动无抑制作用。然而，两种不同的糜蛋白酶样丝氨酸蛋白酶抑制剂TPCK和抑糜酶素，可显著抑制对任何趋化因子的运动。这些结果表明，一种活性糜蛋白酶样丝氨酸蛋白酶对于中性粒细胞对趋化刺激作出反应至关重要。
