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Interleukin-I, Epidermal Growth Factor, and Transforming Growth Factor-ß Exhibit Differential Kinetics on Endothelin-I Synthesis in Amnion Cells

Department of Obstetrics and Gynecology and Laboratory of Perinatal Research, The Ohio State University, College of Medicine, Columbus, Ohio; United States Air Force

Philip Samuels, MD

Peter D. Zimmerman, BS

Department of Obstetrics and Gynecology and Laboratory of Perinatal Research, The Ohio State University, College of Medicine, Columbus, Ohio

Douglas A. Kniss, PhD

Department of Obstetrics and Gynecology and Laboratory of Perinatal Research, The Ohio State University, College of Medicine, Columbus, Ohio kniss.1{at}osu.edu

Objective: To investigate the effects of three cytokines, interleukin-1 (IL-1), epidermal growth factor (EGF), and transforming growth factor-ß (TGF-ß), on the regulation of endothelin-1 (ET-1) mRNA and protein production in human amnion cells.

Methods: Human amnion cells were harvested from uncomplicated pregnancies undergoing elective cesarean delivery at term and grown in primary monolayer culture. Cells were treated with IL-1, EGF, and TGF-ß for dose-response and time course experiments. Northern analysis was used to determine ET-1 mRNA expression, and enzyme-linked immunosorbent assay was used for ET-1 peptide determination.

Results: Interleukin-1, EGF, and TGF-ß induced the expression of ET-1 mRNA and protein in a dose- and time-dependent fashion. The kinetics of ET-1 mRNA production did not differ markedly with respect to the inducing cytokine, but the kinetics of ET-1 protein production was quite different. Interleukin-1 and EGF stimulated a rapid increase in ET-1 that peaked by 24 hours, and the levels declined to just above the detection limit by 72 hours. In contrast, TGF-ß-stimulated cells showed modest ET-1 production at early times (4-24 hours) and then gradually increased and peaked at 72 hours.

Conclusions: Cytokines modulate the expression of ET-1 mRNA and its cognate protein in human amnion cells. The differential kinetics of ET-1 peptide expression in amnion cells suggests that ET metabolism as well as synthesis contribute to the net expression of endothelin in amnion.

Key Words: Cytokines • amnion • endothelin • pregnancy complications

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