Basic fibroblast growth factor release by human coronary artery endothelial cells is enhanced by matrix proteins, 17beta-estradiol, and a PKC signaling pathway.
Basic fibroblast growth factor release by human coronary artery endothelial cells is enhanced by matrix proteins, 17beta-estradiol, and a PKC signaling pathway.
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基质蛋白、17β-雌二醇和 PKC 信号通路可增强人冠状动脉内皮细胞释放碱性成纤维细胞生长因子。
DOI：
10.1006/excr.1998.4243
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发表时间：
1998
影响因子：
3.7
通讯作者：
Schnaper,HW
中科院分区：
文献类型：
--
作者：
Albuquerque,ML;Akiyama,SK;Schnaper,HW
内皮细胞功能是由细胞、细胞外基质（ECM）和可溶性介质之间的相互作用调节的。我们通过检测17β-雌二醇（E2）对在ECM蛋白上培养的人冠状动脉内皮细胞（HCAEC）释放碱性成纤维细胞生长因子（FGF-2）的影响来研究这种相互作用。用E2处理雌激素耗尽的HCAEC 2 h后，通过免疫印迹或ELISA评价条件培养基和细胞层中的FGF-2。释放成纤维细胞生长因子-2到条件培养基增强10倍相比，塑料和进一步的2.4倍E2。随着FGF-2从细胞释放到培养基中的增加，FGF-2的细胞含量相应减少。通过ELISA，FGF-2释放增加406，179，和262%，IV型胶原蛋白，层粘连蛋白，或纤连蛋白，分别。通过ELISA或免疫印迹分析，在I型胶原上培养的HCAEC没有显示E2增强的FGF-2释放。HCAEC释放的乳酸脱氢酶（作为细胞完整性的对照蛋白）未观察到变化。雌激素受体拮抗剂ICI 182，780阻断E2诱导的FGF-2释放，但不阻断基础FGF-2释放。FGF-2释放增加通过环己酰亚胺不敏感途径发生。布雷菲德菌素-A和染料木黄酮均不抑制E2对在纤连蛋白上培养的HCAEC释放FGF-2的增强作用。然而，蛋白激酶C抑制剂calphostin C抑制E2增强的FGF-2释放。这些数据表明，E2增强FGF-2释放的HCAEC上培养的基底膜蛋白在没有创伤。这种作用需要雌激素受体和PKC活性，但不需要新的蛋白质合成，内质网到高尔基体介导的分泌，或蛋白质酪氨酸磷酸化。E2促进FGF-2释放可能有助于雌激素的心脏保护作用。
Endothelial cell function is regulated by interactions among cells, the extracellular matrix (ECM), and soluble mediators. We investigated this interaction by examining the effect of 17β-estradiol (E2) on release of basic fibroblast growth factor (FGF-2) by human coronary artery endothelial cells (HCAEC) cultured on ECM proteins. After estrogen-depleted HCAEC were treated with E2 for 2 h, the conditioned media and cell layers were evaluated by immunoblot or ELISA for FGF-2. Release of FGF-2 into conditioned media was enhanced 10-fold compared to that on plastic and a further 2.4-fold by E2. As FGF-2 release from cells into the media increases, there is a corresponding decrease in the cellular content of FGF-2. By ELISA, FGF-2 release increased 406, 179, and 262%, on type IV collagen, laminin, or fibronectin, respectively. HCAEC cultured on type I collagen did not show E2-enhanced FGF-2 release by ELISA or immunoblot analysis. No changes were noted in HCAEC release of lactate dehydrogenase, tested as a control protein for cellular integrity. The estrogen receptor antagonist ICI182,780 blocked E2-induced, but not basal, FGF-2 release. Increased FGF-2 release occurred via a cycloheximide-insensitive pathway. Neither brefeldin-A nor genistein inhibited E2 enhancement of FGF-2 release by HCAEC cultured on fibronectin. However, the protein kinase C inhibitor calphostin C inhibited the E2-augmented FGF-2 release. These data show that E2 enhances FGF-2 release by HCAEC cultured on basement membrane proteins in the absence of wounding. This action requires the estrogen receptor and PKC activity, but does not require new protein synthesis, endoplasmic reticulum-to-Golgi-mediated secretion, or protein tyrosine phosphorylation. E2-enhanced FGF-2 release could contribute to the cardioprotective effects of estrogen.