Department of Neonatology, University of Leipzig, Leipzig 04103, Germany.
Pediatr Res. 2011 Mar;69(3):200-5. doi: 10.1203/PDR.0b013e3182070ec8.
The effects of estradiol (E2) and progesterone (P) on alveolar epithelial Na+ transport were studied in isolated alveolar epithelial cells from 18- to 19-d GA rat fetuses, grown to confluence in serum-free media supplemented with E2 (0-1 μM) and P (0-2.8 μM). Short-circuit currents (ISC) were measured, showing an increase by E2 and P in a dose-dependent manner. The Na,K-ATPase subunits -α1 and -β1 were detected by Western blotting, but total expression was not significantly altered. Furthermore, all three epithelial Na+ channel (ENaC) subunits -α, -β, and -γ were detected, with trends toward a higher expression in the presence of E2 and P. Real-time PCR revealed an increase of α- and β-ENaC expression but no alteration of γ-ENaC. In addition, the mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR) and Na,K-ATPase-β1 subunit were elevated in the presence of E2 and P. Single-channel patch clamp analysis demonstrated putative highly selective and nonselective cation channels in the analyzed cells, with a higher percentage of responsive patches under the influence of E2 and P. We conclude that E2 and P increased Na+ transport in alveolar epithelial cells by enhancing the expression and activity of ENaC and Na,K-ATPase.
研究了雌二醇(E2)和孕酮(P)对 18-19 天 GA 胎鼠离体肺泡上皮细胞钠离子转运的影响,这些细胞在无血清培养基中生长至汇合,并补充有 E2(0-1 μM)和 P(0-2.8 μM)。测量短路电流(ISC),结果显示 E2 和 P 呈剂量依赖性增加。通过 Western blot 检测到 Na,K-ATPase 亚基-α1 和-β1,但总表达没有明显改变。此外,还检测到所有三种上皮钠离子通道(ENaC)亚基-α、β和-γ,存在 E2 和 P 时表达趋势更高。实时 PCR 显示α-和β-ENaC 表达增加,但 γ-ENaC 没有改变。此外,E2 和 P 存在时囊性纤维化跨膜电导调节因子(CFTR)和 Na,K-ATPase-β1 亚基的 mRNA 表达增加。单通道膜片钳分析表明,在分析的细胞中存在具有高选择性和非选择性阳离子通道的细胞,在 E2 和 P 的影响下,反应性斑块的比例更高。我们得出结论,E2 和 P 通过增强 ENaC 和 Na,K-ATPase 的表达和活性来增加肺泡上皮细胞的钠离子转运。