Human airway trypsin-like protease stimulates human bronchial fibroblast proliferation in a protease-activated receptor-2-dependent pathway
Human airway trypsin-like protease stimulates human bronchial fibroblast proliferation in a protease-activated receptor-2-dependent pathway
复制标题
DOI：
10.1152/ajplung.00098.2005
复制
发表时间：
2006-02-01
影响因子：
4.9
通讯作者：
Yasuoka, S
中科院分区：
文献类型：
--
作者：
Matsushima, R;Takahashi, A;Yasuoka, S
从人气道分泌物中分离出人气道胰蛋白酶样蛋白酶（HAT），并通过免疫组织化学方法定位于支气管上皮细胞。在本研究中，我们研究了HAT是否可以刺激原代人支气管成纤维细胞（HBF）的DNA合成和增殖。HAT显著刺激HBF增殖20 - 55%，与肺肥大细胞类胰蛋白酶（MCT）的促有丝分裂活性水平相似。HAT还刺激HBF中[H-3]胸苷的掺入，并且这种HAT诱导的DNA合成被亮抑酶肽消除。蛋白酶激活受体-2（PAR-2）mRNA在HBF中表达并定位于细胞表面。PAR-2激活肽（AP）也促进DNA合成，HAT和PAR-2 AP诱导受体内化，类似于胰蛋白酶的反应。用抗PAR-2抗体预处理HBF显著抑制HAT和PAR-2 AP诱导的DNA合成。此外，HAT和PAR-2 AP诱导HBF细胞内Ca ~（2+）动员。用胰蛋白酶或PAR-2 AP预处理可使HAT诱导的Ca 2+增加脱敏。特异性MAPK抑制剂U 0126可完全抑制HAT诱导的DNA合成和MAPK磷酸化。HAT和MCT一起作用于HBF DNA合成是累加的，而HAT和胰岛素一起作用于HBF DNA合成是协同的。这些结果表明，HAT通过PAR-2依赖性MEK-MAPK介导的途径刺激支气管气道中的成纤维细胞增殖，并且HAT与涉及成纤维细胞的气道过程有关。
Human airway trypsinlike protease (HAT) was isolated from airway secretions and localized to bronchial epithelial cells by immunohistochemistry. In the present study, we examined whether HAT could stimulate DNA synthesis and proliferation of primary human bronchial fibroblasts (HBF). HAT significantly stimulated the proliferation of HBF by 20 - 55%, a level similar to that of the mitogenic activity of lung mast cell tryptase (MCT). HAT also stimulated the incorporation of [H-3] thymidine in HBF, and this HAT-induced DNA synthesis was abolished by leupeptin. Protease-activated receptor-2 (PAR-2) mRNA was expressed and localized to the cell surface in HBF. PAR-2 activating peptide (AP) also enhanced DNA synthesis, and both HAT and PAR-2 AP induced receptor internalization, similar to the response to trypsin. Pretreatment of HBF with anti-PAR-2 antibody significantly suppressed both HAT and PAR-2 AP-induced DNA synthesis. In addition, HAT and PAR-2 AP induced intracellular Ca2+ mobilization in HBF. The HAT-induced increase in Ca2+ was desensitized by pretreatment with trypsin or PAR-2 AP. U0126, a specific MAPK inhibitor, completely inhibited HAT-induced DNA synthesis as well as HAT-induced phosphorylation of MAPK. The effect of HAT and MCT together was additive, whereas the effect of HAT and insulin together on HBF DNA synthesis was synergistic. These results indicate that HAT stimulates fibroblast proliferation in bronchial airways through a PAR-2-dependent MEK-MAPK mediated pathway and that HAT is linked to airway processes involving fibroblasts.