Deterding R R, Shimizu H, Fisher J H, Shannon J M
Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, Denver, CO 80206.
Am J Respir Cell Mol Biol. 1994 Jan;10(1):30-7. doi: 10.1165/ajrcmb.10.1.8292379.
SP-D is a recently described lung-associated protein that is produced by alveolar type II cells and may function in pulmonary host defenses. Since little is known regarding the hormonal regulation of SP-D, and since the other surfactant proteins (SP-A, SP-B, and SP-C) are known to be regulated by glucocorticoids, we sought to determine the effects of glucocorticoids on SP-D mRNA and protein expression, both in vitro and in vivo, in the fetal rat lung. In vitro experiments were performed on lung explants from fetuses on gestational day 15 or 18. Explants were cultured in serum-free conditions with or without hydrocortisone for 3 days. SP-D mRNA expression was evaluated by Northern blot analysis. SP-D protein expression was analyzed using a polyclonal antibody against SP-D and standard immunohistochemical techniques. The expression of SP-D mRNA increased in fetal day 15 explants but remained unchanged in fetal day 18 explants cultured without the addition of hydrocortisone, compared with in vivo controls. The addition of hydrocortisone resulted in increases in SP-D mRNA expression at both gestational ages. This pattern of SP-D mRNA expression was compared with the expression of the other surfactant proteins and found to be most similar to that of SP-B. In vivo experiments were performed using maternal administration of dexamethasone (1 mg/kg) or an equal volume of saline on fetal days 15, 16, and 17 or on fetal day 17 with sacrifice on fetal day 18.(ABSTRACT TRUNCATED AT 250 WORDS)
表面活性蛋白-D是一种最近被描述的与肺相关的蛋白质,由II型肺泡细胞产生,可能在肺部宿主防御中发挥作用。由于对表面活性蛋白-D的激素调节知之甚少,且已知其他表面活性物质蛋白(表面活性蛋白-A、表面活性蛋白-B和表面活性蛋白-C)受糖皮质激素调节,我们试图确定糖皮质激素在体外和体内对胎鼠肺中表面活性蛋白-D的mRNA和蛋白表达的影响。对妊娠第15天或18天胎儿的肺外植体进行体外实验。将外植体在无血清条件下培养,添加或不添加氢化可的松,培养3天。通过Northern印迹分析评估表面活性蛋白-D的mRNA表达。使用抗表面活性蛋白-D的多克隆抗体和标准免疫组织化学技术分析表面活性蛋白-D的蛋白表达。与体内对照相比,在未添加氢化可的松的情况下培养的妊娠第15天胎儿外植体中,表面活性蛋白-D的mRNA表达增加,而妊娠第18天胎儿外植体中则保持不变。添加氢化可的松导致两个胎龄的表面活性蛋白-D的mRNA表达均增加。将这种表面活性蛋白-D的mRNA表达模式与其他表面活性物质蛋白的表达进行比较,发现与表面活性蛋白-B的表达最相似。体内实验是通过在妊娠第15、16和17天或妊娠第17天给母体注射地塞米松(1mg/kg)或等体积的生理盐水,并在妊娠第18天处死胎儿来进行的。(摘要截短于250字)