Li Wenjiao, Gao Lu, Wang Yan, Duan Tao, Myatt Leslie, Sun Kang
School of Life Sciences, The First Maternal and Fetal Care Hospital, Fudan University, Shanghai, China.
Endocrinology. 2006 May;147(5):2490-5. doi: 10.1210/en.2005-1626. Epub 2006 Feb 9.
Chorion is the most abundant site of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) expression within intrauterine tissues. It is important to study the regulation of 11beta-HSD1 expression in the chorion in terms of local cortisol production during pregnancy. Using real-time PCR and enzyme activity assay, we found that cortisol (1 mum) and IL-1beta (10 ng/ml) for 24 h significantly increased 11beta-HSD1 mRNA expression and reductase activity in cultured human chorionic trophoblasts. A further significant increase of 11beta-HSD1 mRNA expression and reductase activity was observed with cotreatment of cortisol and IL-1beta. To explore the mechanism of induction, 11beta-HSD1 promoter was cloned into pGL3 plasmid expressing a luciferase reporter gene. By transfecting the constructed vector into WISH cells, an amnion-derived cell line, we found that cortisol (1 microM) or IL-1beta (10 ng/ml) significantly increased reporter gene expression. Likewise, an additional increase in reporter gene expression was observed with cotreatment of cortisol and IL-beta. To explore the physiological significance of 11beta-HSD1 induction in the chorion, we studied the effect of cortisol on cytosolic phospholipase A(2) and cyclooxygenase 2 expression. We found that treatment of chorionic trophoblast cells with cortisol (1 microM) induced both cytosolic phospholipase A(2) and cyclooxygenase 2 mRNA expression. We conclude that cortisol up-regulates 11beta-HSD1 expression through induction of promoter activity, and the effect was enhanced by IL-1beta, suggesting that more biologically active glucocorticoids could be generated in the fetal membranes in the presence of infection, which may consequently feed forward in up-regulation of prostaglandin synthesis.
绒毛膜是子宫内组织中11β-羟基类固醇脱氢酶1型(11β-HSD1)表达最丰富的部位。从孕期局部皮质醇生成的角度研究绒毛膜中11β-HSD1表达的调控具有重要意义。通过实时定量PCR和酶活性测定,我们发现,皮质醇(1μM)和白细胞介素-1β(10 ng/ml)作用24小时可显著增加培养的人绒毛膜滋养层细胞中11β-HSD1 mRNA表达和还原酶活性。同时给予皮质醇和白细胞介素-1β时,11β-HSD1 mRNA表达和还原酶活性进一步显著增加。为探究诱导机制,将11β-HSD1启动子克隆到表达荧光素酶报告基因的pGL3质粒中。通过将构建的载体转染到羊膜来源的WISH细胞系中,我们发现皮质醇(1μM)或白细胞介素-1β(10 ng/ml)可显著增加报告基因表达。同样,同时给予皮质醇和白细胞介素-1β时,报告基因表达进一步增加。为探究绒毛膜中11β-HSD1诱导的生理意义,我们研究了皮质醇对胞质磷脂酶A2和环氧化酶2表达的影响。我们发现,用皮质醇(1μM)处理绒毛膜滋养层细胞可诱导胞质磷脂酶A2和环氧化酶2 mRNA表达。我们得出结论,皮质醇通过诱导启动子活性上调11β-HSD1表达,白细胞介素-1β可增强该效应,提示在感染情况下胎膜中可能产生更多具有生物活性的糖皮质激素,这可能进而促进前列腺素合成的上调。