Gööz M, Shaker M, Gööz P, Smolka A J
Gastroenterology and Hepatology Division, Department of Medicine, Medical University of South Carolina, Charleston, SC, USA.
Gut. 2003 Sep;52(9):1250-6. doi: 10.1136/gut.52.9.1250.
and aims: Matrix metalloproteinases (MMPs) are endopeptidases with roles in extracellular matrix remodelling, cell proliferation, and inflammatory processes. We showed previously that Helicobacter pylori infection of human gastric adenocarcinoma (AGS) cells increased epithelial secretion of epithelial MMP-1 and MMP-3 and bacterial secretion of MMP-3-like activity. In the present study, we sought to characterise the role of interleukin (IL)-1beta in H pylori induced secretion of epithelial MMPs.
AGS cells were treated with H pylori and/or IL-1beta. Comparable IL-8 secretory responses (approximately 1700 ng/ml) measured by ELISA were induced by 2.0 ng/ml IL-1beta and by H pylori at a multiplicity of infection (MOI) of 50. The same IL-1beta and H pylori concentrations induced comparable increases in AGS cell caseinolytic activity at 60 kDa. MMP-3 monoclonal antibody immunoblots of AGS cell conditioned media detected immunoreactive bands at 71 kDa and 56 kDa. H pylori (MOI=50-100) induced dose dependent increases in both bands whereas IL-1beta (0.2-2 ng/ml) induced dose dependent increases only in the 71 kDa band, which was identified as a MMP-3/TIMP-3 (tissue inhibitor of metalloproteinases 3) heterodimer. AGS/H pylori conditioned media expressed 24 times more MMP-3 activity than AGS/IL-1beta conditioned media. There was a strong interaction between IL-1beta and H pylori on MMP-3 secretion.
We conclude that IL-1beta induces gastric epithelial cell MMP-3 secretion, contributing to epithelial tissue destruction during H pylori infection. However, other bacterial/host factors are needed to mediate the full gastric epithelial cell MMP-3 secretory response induced by H pylori infection.
基质金属蛋白酶（MMPs）是一类内肽酶，在细胞外基质重塑、细胞增殖及炎症过程中发挥作用。我们之前的研究表明，幽门螺杆菌感染人胃腺癌（AGS）细胞会增加上皮细胞MMP-1和MMP-3的分泌以及细菌MMP-3样活性的分泌。在本研究中，我们试图明确白细胞介素（IL）-1β在幽门螺杆菌诱导上皮细胞MMPs分泌中的作用。
用幽门螺杆菌和/或IL-1β处理AGS细胞。通过酶联免疫吸附测定法（ELISA）测得，2.0 ng/ml的IL-1β和感染复数（MOI）为50的幽门螺杆菌诱导出了相当的IL-8分泌反应（约1700 ng/ml）。相同浓度的IL-1β和幽门螺杆菌在60 kDa处诱导出了相当的AGS细胞酪蛋白溶解活性增加。AGS细胞条件培养基的MMP-3单克隆抗体免疫印迹检测到71 kDa和56 kDa处有免疫反应条带。幽门螺杆菌（MOI = 50 - 100）诱导这两条带呈剂量依赖性增加，而IL-1β（0.2 - 2 ng/ml）仅诱导71 kDa条带呈剂量依赖性增加，该条带被鉴定为MMP-3/TIMP-3（金属蛋白酶组织抑制剂3）异二聚体。AGS/幽门螺杆菌条件培养基的MMP-3活性比AGS/IL-1β条件培养基高24倍。IL-1β和幽门螺杆菌在MMP-3分泌上存在强烈的相互作用。
我们得出结论，IL-1β诱导胃上皮细胞MMP-3分泌，在幽门螺杆菌感染期间促成上皮组织破坏。然而，还需要其他细菌/宿主因子来介导幽门螺杆菌感染诱导的完整胃上皮细胞MMP-3分泌反应。
