Production of matrix metalloproteinase-9 in lipopolysaccharide-stimulated human amnion occurs through an autocrine and paracrine proinflammatory cytokine-dependent system
Production of matrix metalloproteinase-9 in lipopolysaccharide-stimulated human amnion occurs through an autocrine and paracrine proinflammatory cytokine-dependent system
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DOI：
10.1095/biolreprod.102.004721
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发表时间：
2002-12-01
影响因子：
3.6
通讯作者：
Vadillo-Ortega, F
中科院分区：
文献类型：
--
作者：
Arechavaleta-Velasco, F;Ogando, D;Vadillo-Ortega, F
本研究的目的是确定存在的自分泌/旁分泌调节基质金属蛋白酶-9（MMP-9）的表达的促炎细胞因子介导的人胎膜。从分娩前接受剖腹产的妇女获得的胎膜被手工分离成羊膜和绒毛膜层并保持在培养物中。这些外植体;用肿瘤坏死因子α（TNF α）、白细胞介素-1 β（IL-1 β）和单独的脂多糖（LPS）或LPS与抗TNF α或抗IL-1 β中和抗体刺激。通过酶谱法评估培养基中的proMMP-9水平。采用酶联免疫吸附法检测LPS刺激后IL-1 β、TNF α和基质金属蛋白酶组织抑制剂-1（TIMP-1）的含量。在LPS、TNF α和IL-10刺激羊膜后，ProMMP-9活性上调。羊膜中由LPS刺激引起的proMMP-9活性增加被TNF α中和抗体阻断，但抗IL-10不能阻断。在绒毛膜中未观察到LPS、TNF α或IL-10对proMMP-9表达的显著影响;然而，当用LPS刺激时，绒毛膜产生两种细胞因子。相反，在LPS存在下孵育的所有培养物中，TMP-1水平保持不变。因此，这些数据表明，proMMP-9是由羊膜产生的，而不是绒毛膜响应于LPS。由于抗TNF α中和抗体抑制羊膜中的proMMP-9活性，TNF α似乎以自分泌方式上调羊膜产生的proMMP-9。同时，绒毛膜产生的TNF α和IL-10可能以旁分泌方式上调子宫内膜proMMP-9的产生。
The objective of this study was to determine the presence of autocrine/paracrine regulation of matrix metalloproteinase-9 (MMP-9) expression mediated by proinflammatory cytokines in human fetal membranes. Fetal membranes obtained from women who underwent cesarean delivery before labor were manually separated into amnion and chorion layers and maintained in culture. These explants; were stimulated with tumor necrosis factor alpha (TNFalpha), interleukin-1beta (IL-1beta), and either lipopolysaccharide (LPS) alone or LPS with anti-TNFalpha or anti-IL-1beta-neutralizing antibodies. Levels of proMMP-9 in culture media were evaluated by zymography. Enzyme-linked immunosorbant assay was performed to measure the quantity of IL-1beta, TNFalpha, and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) after LPS stimulation. ProMMP-9 activity was upregulated after stimulation of the amnion by LPS, TNFalpha, and IL-10. The increased activity of proMMP-9 resulting from LPS stimulation in the amnion was blocked by the addition of TNFalpha neutralizing antibody but not with anti-IL-10. No significant effect of LPS, TNFalpha, or IL-10 on proMMP-9 expression was observed in the chorion; however, the chorion produced both cytokines when stimulated with LPS. In contrast, TMP-1 levels remained unchanged in all cultures incubated in the presence of LPS. Therefore, these data indicate that proMMP-9 is produced by the amnion but not the chorion in response to LPS. Because anti-TNFalpha-neutralizing antibody inhibits proMMP-9 activity in the amnion, TNFalpha appears to upregulate proMMP-9 production by the amnion in an autocrine fashion. Meanwhile, TNFalpha and IL-10 produced by the chorion may upregulate amnionic proMMP-9 production in a paracrine manner.