Recombinant human elafin protects airway epithelium integrity during inflammation
Recombinant human elafin protects airway epithelium integrity during inflammation
复制标题
重组人弹性蛋白在炎症过程中保护气道上皮的完整性
DOI：
10.1007/s11033-009-9865-z
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发表时间：
2010-07-01
影响因子：
2.8
通讯作者：
Yang, Jie
中科院分区：
文献类型：
--
作者：
Li, Qi;Zhou, Xiang Dong;Yang, Jie
Elafin是一种抗蛋白酶，可能保护肺上皮细胞免受炎症损伤。本研究旨在探讨重组人弹力素保护A549细胞完整性免受炎症攻击的分子机制。分别用真核表达载体pEGFP-N1-Elafin和阴性对照载体pEGFP-N1转染A549气道上皮细胞。将细胞与多形核中性粒细胞（PMN）共孵育，然后用脂多糖（LPS）刺激。结果显示，pEGFP-N1-Elafin转染细胞经LPS刺激后，中性粒细胞弹性蛋白酶（NE）活性显著降低，同时elafin mRNA表达和蛋白表达水平显著升高;与pEGFP-N1-Elafin转染组相比，pEGFP-N1-Elafin转染组NE活性显著升高，elafin表达显著降低（P< 0.05）。在转染pEGFP-N1的细胞中，LPS抑制紧密连接蛋白紧密连接蛋白ZO-1的表达，而在重组pEGFP-N1-Elafin细胞中，LPS对ZO-1表达无明显影响。LPS刺激后，pEGFP-N1转染细胞的胶原粘附能力明显减弱（P< 0.01），而重组pEGFP-N1-Elafin细胞的胶原粘附能力无显著性差异（P> 0.05）。提示弹力素可通过保护气道上皮细胞，增强气道抗炎能力，维持气道上皮完整性。
Elafin, an antiprotease, is likely to protect pulmonary epithelial cells from inflammatory damages. We aimed to explore the molecule mechanisms of recombinant human elafin on protecting A549 cells integrity against inflammatory assault. We transfected A549 airway epithelial cells with eukaryotic expression vector pEGFP-N1-Elafin and negative control vector pEGFP-N1, respectively. Cells were co-incubated with polymorphonuclear neutrophils (PMN) and then were stimulated with lipopolysaccharide (LPS). Results revealed that, in pEGFP-N1-Elafin transfected cells, neutrophil elastase (NE) activity significantly decreased after LPS stimulation, accompanied with elevated elafin mRNA level and protein production, whereas in cells transfected with pEGFP-N1, NE activity was higher and elafin expression was lower, compared with pEGFP-N1-Elafin transfected group (P< 0.05). In pEGFP-N1 transfected cells, LPS suppressed tight junctions protein zonula occludens-1 (ZO-1) production, while in recombinant pEGFP-N1-Elafin cells, LPS did not cause significantly decrease of ZO-1, compared with normal control cells. LPS stimulation also significantly weakened the collagen adhesion capability of pEGFP-N1 transfected cells (P< 0.01), but there was no significant difference in recombinant pEGFP-N1-Elafin cells (P> 0.05). These results suggest that elafin can maintain airway epithelium integrity by protecting airway epithelial cells and enhancing the anti-inflammatory capability of airway.