Evidence of in vitro differential secretion of 72 and 92 kDa type IV collagenases after selective exposure to lipopolysaccharide in human fetal membranes
Evidence of in vitro differential secretion of 72 and 92 kDa type IV collagenases after selective exposure to lipopolysaccharide in human fetal membranes
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DOI：
10.1093/molehr/gam025
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发表时间：
2007-05-01
影响因子：
4
通讯作者：
Zaga-Clavellina, Verónica
中科院分区：
文献类型：
--
作者：
Garcia-Lopez, Guadalupe;Vadillo-Ortega, Felipe;Zaga-Clavellina, Verónica
绒毛膜羊膜破裂合并宫内感染与细胞外基质（ECM）降解有关，这可以解释局部形态学变化。我们使用了一种培养系统，其中绒毛膜羊膜形成两个独立的腔室，允许选择性刺激羊膜（AMN）和/或绒毛膜蜕膜（CHD）区域。将脂多糖（500 ng/ml）添加到AMN和/或CHD中;通过酶联免疫吸附测定（ELISA）和酶谱法测定两个隔室中基质金属蛋白酶（MMP）-2和MMP-9的分泌和明胶分解活性。采用酶联免疫吸附法（ELISA）检测TIMP-1、TIMP-2和TIMP-4的分泌。这两种金属蛋白酶在组织切片中进行免疫定位。所有刺激方式在CHD中诱导相似的proMMP-2和proMNIP-9分泌模式，浓度分别为2.49 ng/ml和90.91 pg/ml; ANIN未显示出显著变化。这两种酶的活性形式没有改变任何刺激方式。TIMP-1、TIMP-2和TIMP-41的分泌无明显变化（P = 0.41）。刺激后ECM降解和结构紊乱明显。proMNIP-2和proMMP-9主要在CHD中分泌，与组织相关的活性形式的存在以及TIMPs分泌的微小变化可能有利于ECM降解，并解释了与绒毛膜羊膜病理性破裂相关的弱化和变薄。
Premature rupture of chorioamniotic membranes complicated with intrauterine infection has been associated to degradation of extracellular matrix (ECM), which could explain local morphological changes. We used a culture system in which the chorioamniotic membranes form two independent chambers, allowing for the selective stimulation of either the amnion (AMN) and/ or the choriodecidua (CHD) regions. Lipopolysaccharide (500 ng/ml) was added to the AMN and/or the CHD; secretions and gelatinolytic activity of matrix metalloproteinase (MMP)-2 and MMP-9 were measured in both compartments by enzyme-linked immunosorbent assay (ELISA) and zymography. Secretions of TIMP-1, TIMP-2 and TIMP-4 were measured by ELISA. Both metalloproteinases were immunolocalized in tissue sections. All stimulation modalities induced a similar proMMP-2 and proMNIP-9 secretion pattern in the CHD with concentrations of 2.49 ng/ml and 90.91 pg/ml, respectively; the ANIN showed no significant changes. The active forms of both enzymes did not change with any stimulation modality. TIMP-1, TIMP-2 and TIMP-41 secretions remained without significant changes (P = 0.41). ECM degradation and structural disarrangement were evident after stimulation. Secretion of proMNIP-2 and proMMP-9 mainly in the CHD, presence of active forms associated to the tissue and minor changes in TIMPs secretion could favor ECM degradation and explain the weakening and thinning associated with the pathological rupture of chorioamniotic membranes.