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Histochemical Analyses of Altered Fetal Lung Development Following Single vs Multiple Courses of Antenatal Steroids

Zarah J. Pua, Barbara S. Stonestreet, Anne Cullen, Aliakbar Shahsafaei, Grazyna B. Sadowska and Mary E. Sunday

Department of Medicine and Department of Pathology, Children's Hospital and Harvard Medical School, Boston, Massachusetts (ZJP,AC,MES); Department of Pediatrics, Women's & Infants' Hospital and Brown University, Providence, Rhode Island (BSS,GBS); and Department of Pathology, Brigham & Women's Hospital, Boston, Massachusetts (AS,MES)

Correspondence and present address: Mary E. Sunday, MD, PhD, Duke University Medical Center, Box 3712, Durham, NC 27708. E-mail: mary.sunday{at}duke.edu

A single course of antenatal steroids is widely used during preterm labor to promote fetal lung maturation. However, little is known regarding efficacy and safety of multiple courses of antenatal steroids. In animal models and clinical trials, treatment with glucocorticoids can inhibit growth. The present study of single- vs multiple-course steroids in pregnant ewes analyzes the effects of steroids vs placebo on fetal lung histopathology. Single-course groups received dexamethasone (Dex) 6 mg or normal saline every 12 hr for 48 hr at 104–106 days of gestation (term = 150 days). Multiple-course groups received the first course at 76–78 days; this was repeated weekly for 5 weeks. At 108 days, lungs were analyzed using immunohistochemistry for -smooth muscle actin, a myofibroblast marker and proliferating cell nuclear antigen. Cell injury/death was evaluated using TdT-mediated dUTP digoxigenin nick end labeling (TUNEL) analysis. Although fetal growth was restricted by either single or multiple courses of Dex, alveolar development was accelerated as measured by mean linear intercepts. Alveolar walls were thinner, developing septa were longer, and septal myofibroblasts were increased for both Dex groups compared with controls. Cell proliferation increased following multiple steroid courses, especially in the distal parenchyma, with a corresponding decrease in apoptosis. These observations suggest that Dex promotes alveolarization, whether given in single or multiple courses.

(J Histochem Cytochem 53:1469–1479, 2005)

Key Words: glucocorticoids • sheep • immunohistochemistry • morphometry • smooth muscle actin • proliferating cell nuclear • antigen • apoptosis

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