Wang Chung-An, Liu Yen-Chun, Du Shin-Yi, Lin Chio-Wen, Fu Hua-Wen
Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Biochem Biophys Res Commun. 2008 Dec 5;377(1):52-6. doi: 10.1016/j.bbrc.2008.09.072. Epub 2008 Sep 25.
Helicobacter pylori infection induces acute and chronic inflammation and plays a key role in gastric mucosal diseases. H. pylori neutrophil-activating protein (HP-NAP), one of its virulence factors, induces not only chemotactic but also oxidative burst responses of neutrophils. Activated neutrophils use myeloperoxidase (MPO) to generate many cytotoxic oxidants, which might result in gastric mucosal injury. In this study, we evaluated whether HP-NAP could promote MPO release from human neutrophils. Recombinant HP-NAP expressed in Escherichia coli was purified by two sequential gel filtration chromatographies and then subjected to syringe filtration for endotoxin removal. The purified recombinant HP-NAP assembles into a multimer with a secondary structure of the typical alpha-helix. In addition to stimulating the production of reactive oxygen species, HP-NAP is able to induce the secretion of MPO in human neutrophils. The increased MPO release from neutrophils induced by HP-NAP may be related to the pathogenesis of H. pylori-associated gastritis.
幽门螺杆菌感染会引发急性和慢性炎症，在胃黏膜疾病中起关键作用。幽门螺杆菌中性粒细胞激活蛋白（HP-NAP）是其毒力因子之一，不仅能诱导中性粒细胞的趋化反应，还能引发其氧化爆发反应。被激活的中性粒细胞利用髓过氧化物酶（MPO）生成许多细胞毒性氧化剂，这可能导致胃黏膜损伤。在本研究中，我们评估了HP-NAP是否能促进人中性粒细胞释放MPO。在大肠杆菌中表达的重组HP-NAP通过两次连续的凝胶过滤色谱法进行纯化，然后进行注射器过滤以去除内毒素。纯化后的重组HP-NAP组装成具有典型α-螺旋二级结构的多聚体。除了刺激活性氧的产生外，HP-NAP还能够诱导人中性粒细胞分泌MPO。HP-NAP诱导中性粒细胞释放MPO增加可能与幽门螺杆菌相关性胃炎的发病机制有关。
