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Production of Oxytocin Receptor and Cytokines in Primary Uterine Smooth Muscle Cells Cultivated Under Inflammatory Conditions

Ulrike Tretzmüller

Mathias Brunbauer, MD

Andrea Kaider, MsC

Peter Husslein, MD

Departments of Obstetrics and Gynecology and Medical Computersciences, General Hospital, University of Vienna, Vienna, Austria

Martin Knöfler, PhD

Departments of Obstetrics and Gynecology and Medical Computersciences, General Hospital, University of Vienna, Vienna, Austria; Department of Obstetrics and Gynecology, University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria martin.knoefler{at}akh-wien.ac.at

Objective: We studied the production of the oxytocin receptor and interleukins in human uterine smooth muscle cells cultured in vitro in the presence of cytokines that were shown to be elevated in gestational diseases such as intrauterine infections and chorioamnionitis.

Methods: Human uterine smooth muscle cells were cultured in the absence or presence of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor (TNF), or lipopolysaccharide (LPS). Additionally, cells were cultivated under hypoxic conditions (3.5% oxygen). After 6, 12, 24, and 48 hours of incubation, oxytocin receptor mRNA was measured from total RNA using quantitative, competitive reverse transcriptase-polymerase chain reaction. Secreted cytokines (IL-1ß, IL-6, or IL-8) were quantitated from supernatants after 6, 12, 24, and 48 hours of stimulation by commercially available enzyme-linked immunosorbent assay.

Results: In nonstimulated cultures basal secretion of IL-1ß, IL-6, and IL-8 was detectable. Supplementation of IL-1ß induced a statistically significant decrease in oxytocin receptor mRNA abundance, whereas IL-6, TNF, LPS, or hypoxia did not significantly affect oxytocin receptor gene expression. The cytokines IL-1 and TNF induced IL-6 and IL-8 release, whereas secretion of the two interleukins was not altered in the presence of LPS or hypoxia. Expression of IL-1ß was not significantly induced under inflammatory or hypoxic culture conditions.

Conclusion: The constitutive and cytokine-inducible expression of interleukins from uterine smooth muscle cells suggests that the myometrium may contribute to the overall production of inflammatory mediators in the uterus that are thought to govern term or infection-induced preterm labor. Down-regulation of the oxytocin receptor under IL-1ß in myometrial cells may indicate that initiation and maintenance of labor could be partially limited under severe inflammatory conditions such as chorioamnionitis.

Key Words: Uterine smooth muscle cells • oxytocin receptor • inducible cytokine expression • chorioamnionitis

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