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首页> 外文期刊>Journal of reproduction and fertility >Production of matrix metalloproteinase 1 (interstitial collagenase) and matrix metalloproteinase 2 (gelatinase A: 72 kDa gelatinase) by ovine endometrial cells in vitro: different regulation and preferential expression by stromal fibroblasts
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Production of matrix metalloproteinase 1 (interstitial collagenase) and matrix metalloproteinase 2 (gelatinase A: 72 kDa gelatinase) by ovine endometrial cells in vitro: different regulation and preferential expression by stromal fibroblasts

机译：绵羊子宫内膜细胞体外产生基质金属蛋白酶1（间质胶原酶）和基质金属蛋白酶2（明胶酶A：72 kDa明胶酶）：基质成纤维细胞的不同调控和优先表达

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Ovine endometrial cells (epithelial plus stromal), prepared from ovariectomized ewes treated with oestrogen and progesterone to mimic the luteal phase of the oestrous cycle were maintained in serum-free medium for 48 h in the presence or absence of phorbol myristate acetate (PMA, 100 nmol l?1), a known stimulus for production of matrix metalloproteinases (MMP) in other cells. Matrix metalloproteinase-1 (MMP-1, interstitial collagenase) and matrix metalloproteinase-2 (MMP-2, gelatinase A) activities were expressed by the cells in the absence of PMA; most were in the latent form and required activation by (4-aminophenyl) mercuric acetate (APMA). Exposure to PMA over 48 h resulted in a significant increase in MMP-1 activity but only a modest and nonsignificant increase in MMP-2 activity. Gelatin zymography demonstrated that proMMP-2 (72 kDa) was produced by both PMA-treated and untreated cells and an active form of 67 kDa was also present. Immunolocalization of MMP-1 and MMP-2 was seen within the cells following treatment with monensin. Highly purified epithelial and stromal cells were similarly cultured and analysis of the conditioned medium showed that MMP-1 and MMP-2 were produced predominantly by stromal rather than epithelial cells. Thus, both MMP-1, which degrades interstitial collagens, and MMP-2, an important enzyme for degradation of type IV and V collagens, are synthesized and released by ovine endometrial stromal cells in culture, but MMP-1 is produced primarily upon stimulation, whereas MMP-2 production is constitutive. It is postulated that these enzymes have important roles in endometrial remodelling and implantation.
机译：在有或没有佛波肉豆蔻酸酯醋酸盐（PMA，100）的情况下，将由经雌激素和孕酮处理的去卵巢母羊模拟的雌性黄体期模拟的子宫内膜细胞（上皮+基质）在无血清培养基中保持48 h。 （nmol 1），一种在其他细胞中产生基质金属蛋白酶（MMP）的已知刺激物。在不存在PMA的情况下，细胞表达基质金属蛋白酶-1（MMP-1，间质胶原酶）和基质金属蛋白酶-2（MMP-2，明胶酶A）的活性。大多数为潜伏形式，需要被（4-氨基苯基）乙酸汞（APMA）激活。在48小时内暴露于PMA会导致MMP-1活性显着增加，但MMP-2活性仅适度且无显着增加。明胶酶谱显示，proMMP-2（72 kDa）由PMA处理和未处理的细胞共同产生，并且还存在67 kDa的活性形式。用莫能菌素处理后，在细胞内发现了MMP-1和MMP-2的免疫定位。相似地培养高度纯化的上皮和基质细胞，对条件培养基的分析表明，MMP-1和MMP-2主要由基质细胞而非上皮细胞产生。因此，降解间质胶原的MMP-1和降解IV型和V型胶原的重要酶MMP-2都是由培养的绵羊子宫内膜基质细胞合成并释放的，但是MMP-1主要是在刺激下产生的，而MMP-2的生产是构成性的。假定这些酶在子宫内膜重塑和植入中具有重要作用。

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