Matrix metalloproteinase and matrix metalloproteinase inhibitor expression in endometrial stromal cells during progestin-initiated decidualization and menstruation-related progestin withdrawal
Matrix metalloproteinase and matrix metalloproteinase inhibitor expression in endometrial stromal cells during progestin-initiated decidualization and menstruation-related progestin withdrawal
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DOI：
10.1210/en.139.11.4607
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发表时间：
1998-11-01
期刊：
影响因子：
4.8
通讯作者：
Schatz, F
中科院分区：
文献类型：
--
作者：
Lockwood, CJ;Krikun, G;Schatz, F
孕激素（E）在体内和体外引发人子宫内膜基质细胞（HESC）的蜕膜化作用。评价在对照培养基中以及在补充有E或合成的孕激素醋酸甲羟孕酮（P）或E + P的培养基中孵育的汇合HESC中的基质金属蛋白酶（MMP）表达。用特异性ELISA进行的测量表明，基础pro-MMP-1输出不受E的影响，而E + P诱导几种蜕膜化相关标志物的表达，对HESC分泌的pro-MMP-1水平产生时间依赖性抑制。通过北方分析确定，P而不是E降低了MMP-1信使RNA（mRNA）的稳态水平，这与HESC中MMP-1蛋白表达的雷公藤红素抑制一致。相比之下，MMP-2和MMP抑制剂TIMP-1的mRNA水平没有被P或E改变。类固醇戒断研究表明，在用E + P孵育HESC抑制MMP-1表达后，4天暴露于抗白细胞介素RU 486与维持在E + P中的培养物相比，米非司酮在条件培养基中显著上调MMP-1水平数倍。自由对照培养基需要更长的停药期以达到与RU 486诱发的调节作用相当的调节作用。通过用特异性MMP-1抗体进行免疫印迹分析来验证ELISA测量，所述特异性MMP-1抗体在约50 kDa的预期迁移率处显示条带的相应变化。此外，北方分析显示MMP-1 mRNA水平的平行变化，而MMP-2和TIMP-1 mRNA水平均不受添加或撤销类固醇的调节。在培养的HESC中观察到的受调节的MMP-1表达和组成型MMP-2表达之间的对比与所证实的MMP-1启动子上存在MMP-2启动子中不存在的调节元件如AP-1和PEA-3一致。将HESC中的这些体外变化外推到体内子宫内膜事件表明：1）E和黄体酮抑制MMP-1表达将稳定血管周围子宫内膜ECM，以防止植入滋养层血管内入侵期间局部出血; 2）类固醇戒断引起的MMP-1表达增强将介导子宫内膜ECM降解，导致月经期间功能层脱落。
Estradiol(E) primes human endometrial stromal cells (HESCs) for the decidualizing effects of progesterone in vivo and in vitro. Matrix metalloproteinase (MMP) expression was evaluated in confluent HESCs incubated in control medium, and in medium supplemented with either E, or the synthetic progestin medroxyprogesterone acetate (P), or E + P. Measurements with a specific ELISA indicated that basal pro-MMP-l output was unaffected by E, whereas E + P, which induces the expression of several decidualization-related markers, produced a time-dependent inhibition in HESC-secreted levels of pro-MMP-1. Consistent with progestin inhibition of MMP-1 protein expression in the HESCs, P but not E, reduced steady state levels of MMP-1 messenger RNA (mRNA) as determined by Northern analysis. By contrast, mRNA levels for MMP-2 and the MMP inhibitor TIMP-1 were not altered by either P or E. Steroid withdrawal studies indicated that after MMP-1 expression was suppressed by incubation of the HESCs with E + P, 4 days of exposure to the antiprogestin RU 486 (mifepristone) significantly up-regulated MMP-1 levels in the conditioned medium by severalfold compared with cultures maintained in E + P. The change to steroid-free control medium required a more prolonged period of withdrawal to attain up regulatory effects that were comparable with those evoked by RU 486. The ELISA measurements were validated by immunoblot analysis with a specific MMP-1 antibody, which showed corresponding changes in a band at the expected mobility of about 50 kDa. Moreover, Northern analysis revealed parallel changes in MMP-1 mRNA levels, whereas neither MMP-2 nor TIMP-1 mRNA levels were modulated by adding or withdrawing steroids. The contrast between regulated MMP-1 expression and constitutive MMP-2 expression observed in the cultured HESCs is consistent with the demonstrated presence on the MMP-1 promoter of regulatory elements such as AP-1 and PEA-3 that are absent from the MMP-2 promoter. Extrapolation of these in vitro changes in HESCs to in vivo endometrial events suggests that: 1) inhibition of MMP-1 expression by E and progesterone would stabilize the perivascular endometrial ECM to prevent local hemorrhage during endovascular invasion by the implanting trophoblast; 2) enhanced expression of MMP-1 evoked by steroid withdrawal would mediate endometrial ECM degradation leading to sloughing of the functional layer during menstruation.