Estrogen decrease in tight junctional resistance involves matrix-metalloproteinase-7-mediated remodeling of occludin
Estrogen decrease in tight junctional resistance involves matrix-metalloproteinase-7-mediated remodeling of occludin
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DOI：
10.1210/en.2006-1120
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发表时间：
2007-01-01
期刊：
影响因子：
4.8
通讯作者：
Gorodeski, George I.
中科院分区：
文献类型：
--
作者：
Gorodeski, George I.
雌激素通过雌激素受体α介导的闭合蛋白重塑调节紧密连接阻力。这项研究的目的是了解所涉及的机制。用人正常阴道-宫颈上皮细胞进行的实验表明，人正常阴道-宫颈上皮细胞组成性地分泌基质金属蛋白酶-7（MMP-7）到管腔溶液中，MMP-7是产生雌激素、降低紧密连接阻力和重构闭合蛋白所必需的。用雌激素处理刺激细胞内proMMP-7的活化并增加活化MMP-7形式的分泌。MMP-7 mRNA和蛋白的稳态水平不受雌激素的影响。雌激素调节MMP-7的磷酸化，但这些变化很可能是继发于细胞MMP-7质量的变化。雌激素增加MMP-7与高尔基体蛋白GPP 130的共免疫反应性。衣霉素和布雷菲德菌素-A对细胞MMP-7没有影响，但莫能菌素（高尔基体交通抑制剂）阻断了雌激素的作用，表明雌激素位点位于高尔基体系统。雌激素增加广义的分泌活动，包括腔胞吐的多糖。然而，雌激素增加MMP-7与突触体相关蛋白的25 kDa的顶端膜，提示可溶性N-乙基马来酰亚胺敏感的融合因子附着蛋白受体促进的MMP-7的胞吐。用囊泡ATP酶抑制剂巴弗洛霉素A1处理抑制MMP-7的活化。这些数据表明，雌激素在高尔基体水平上调细胞内MMP-7的活化，并通过可溶性N-乙基马来酰亚胺敏感融合因子附着蛋白受体依赖性胞吐作用增加活化MMP-7的分泌。另一方面，雌激素酸化的管腔溶液将趋于碱化胞吐囊泡，并可能导致MMP-7的活化降低。这些机制协同作用可能是重要的调节和控制雌激素调节细胞旁通透性在体内。
Estrogen modulates tight junctional resistance through estrogen receptor-alpha-mediated remodeling of occludin. The objective of the study was to understand the mechanisms involved. Experiments using human normal vaginal-cervical epithelial cells showed that human normal vaginal-cervical epithelial cells secrete constitutively matrix-metalloproteinase-7( MMP-7) into the luminal solution and that MMP-7 is necessary and sufficient to produce estrogen decrease of tight junctional resistance and remodeling of occludin. Treatment with estrogen stimulated activation of the proMMP-7 intracellularly and augmented secretion of the activated MMP-7 form. Steady-state levels of MMP-7 mRNA and protein were not affected by estrogen. Estrogen modulated phosphorylation of the MMP-7, but the changes were most likely secondary to changes in cellular MMP-7 mass. Estrogen increased coimmunoreactivity of MMP-7 with the Golgi protein GPP130. Tunicamycin and brefeldin-A had no effect on cellular MMP-7 but monensin ( inhibitor of Golgi traffic) blocked estrogen effects, suggesting estrogen site of is at the Golgi system. Estrogen increased generalized secretory activity, including of luminal exocytosis of polycarbohydrates. However, estrogen increased coimmunoreactivity of MMP-7 with synaptosomal-associated protein of 25 kDa in apical membranes, suggesting soluble N-ethylmaleimide sensitive fusion factor attachment protein receptor-facilitated exocytosis of MMP-7. Treatment with the vesicular-ATPase inhibitor bafilomycin A1 inhibited activation of MMP-7. These data suggest that estrogen upregulates activation of the MMP-7 intracellularly, at the level of Golgi, and augments secretion of activated MMP-7 through soluble N-ethylmaleimide sensitive fusion factor attachment protein receptor-dependent exocytosis. On the other hand, estrogen acidification of the luminal solution would tend to alkalinize exocytotic vesicles and may lead to decreased activation of the MMP-7. These mechanisms acting in concert could be important for regulation and control of estrogen modulation of paracellular permeability in vivo.