Grant-Tschudy Katherine S, Wira Charles R
Department of Physiology, Dartmouth Medical School, Lebanon, NH 03756-0001, USA.
Immunology. 2005 May;115(1):99-107. doi: 10.1111/j.1365-2567.2005.02134.x.
Oestradiol-17beta (Oe(2)) stimulates uterine epithelial cell proliferation and is critical for normal uterine differentiation and secretory function. Oe(2) can act directly on the epithelium via the epithelial oestrogen receptor (OR) or indirectly via the OR-positive underlying stroma. A primary role for epithelial-stromal interactions has been established for mediating steroid hormone action in the uterus. This study was undertaken to determine the mode of Oe(2) action in regulating epithelial cell cytokine release in the uterus. Mouse uterine epithelial and stromal cells were isolated and cultured separately. Transepithelial resistance (TER) was monitored with an EVOM voltohmmeter to determine monolayer polarity and integrity. Epithelial cells grown alone or in coculture with stromal cells were treated with Oe(2). Supernatants collected were assayed for transforming growth factor-beta (TGF-beta) and tumour necrosis factor-alpha (TNF-alpha) by bioassay and enzyme-linked immunosorbent assay, respectively. While Oe(2) treatment of epithelial cells led to a significant decrease in TER, the amount of TNF-alpha released was not altered. However, when epithelial cells were cocultured with stromal cells and treated with Oe(2), apical TNF-alpha release was significantly decreased, compared to cells not treated with hormone. As determined by oestrogen receptor antagonist studies, Oe(2) primed epithelial cells for the action of the stromal paracrine factor(s). In contrast, TGF-beta release by epithelial cells was not affected by Oe(2) when grown alone or in the presence of stromal cells. These studies indicate that Oe(2) has both direct and indirect effects on the uterine epithelium. While epithelial monolayer integrity is directly influenced by Oe(2), TNF-alpha release in response to Oe(2) is dependent on the presence of stromal cells, indicating that paracrine communication is necessary for steroid regulation of some but not all cytokines.
17β-雌二醇（E₂）可刺激子宫上皮细胞增殖，对子宫的正常分化和分泌功能至关重要。E₂可通过上皮雌激素受体（ER）直接作用于上皮细胞，或通过ER阳性的下层基质间接发挥作用。上皮-基质相互作用在介导子宫类固醇激素作用方面的主要作用已得到确立。本研究旨在确定E₂在调节子宫上皮细胞细胞因子释放中的作用模式。将小鼠子宫上皮细胞和基质细胞分离并分别培养。用EVOM电压电阻仪监测跨上皮电阻（TER），以确定单层细胞的极性和完整性。单独培养或与基质细胞共培养的上皮细胞用E₂处理。收集的上清液分别通过生物测定法和酶联免疫吸附测定法检测转化生长因子-β（TGF-β）和肿瘤坏死因子-α（TNF-α）。虽然用E₂处理上皮细胞导致TER显著降低，但TNF-α的释放量未改变。然而，当上皮细胞与基质细胞共培养并用E₂处理时，与未用激素处理的细胞相比，顶端TNF-α的释放显著降低。通过雌激素受体拮抗剂研究确定，E₂使上皮细胞对基质旁分泌因子的作用产生预适应。相反，上皮细胞单独生长或在有基质细胞存在的情况下，TGF-β的释放不受E₂的影响。这些研究表明，E₂对子宫上皮细胞有直接和间接影响。虽然上皮单层细胞的完整性直接受E₂影响，但E₂诱导的TNF-α释放依赖于基质细胞的存在，这表明旁分泌通讯对于某些但不是所有细胞因子的类固醇调节是必要的。
