GENE-REGULATION OF INTERLEUKIN-1-BETA, INTERLEUKIN-1 RECEPTOR-TYPE-I,AND PLASMINOGEN-ACTIVATOR INHIBITOR-1 AND INHIBITOR-2 IN HUMAN GRANULOSA-LUTEAL CELLS

Objective: To investigate the regulation of messenger ribonucleic acid (mRNA) levels of interleukin-l beta (IL-1 beta), interleukin-l (IL-1) receptor type 1, and plasminogen activator (PA) inhibitor-1 and -2 in cumulus cells, granulosa-luteal cells, and macrophage-depleted granul osa-luteal cells obtained from human preovulatory follicles. Design: P rospective longitudinal study. Setting, Patients: Patients undergoing assisted reproductive technologies (ART) in the Department of Gynecolo gy and Obstetrics, Stanford University, Stanford, California. Interven tions: Cumulus cells and granulosa-luteal cells were collected by ultr asound-guided transvaginal aspiration at the time of ART. Main Outcome Measures: Northern blot analysis of mRNA levels of IL-1 beta, IL-1 re ceptor type 1, PA inhibitor-1 and -2 in cumulus cells, granulosa-lutea l cells and macrophage-depleted granulosa-luteal cells, and indirect i mmunocytochemical analysis of the IL-1 system and macrophages in granu losa-luteal cell preparations were performed. Results: Interleukin-1 b eta mRNA levels in uncultured cumulus cells were less than those of un cultured granulosa-luteal cells with no differences in IL-1 receptor t ype 1 mRNA levels between these two cell types. Granulosa-luteal cell IL-1 receptor type 1 mRNA levels were expressed constitutively through out 24 hours of culture with no effect by hCG, whereas IL-1 beta mRNA levels increased within 6 hours, and then remained elevated for 24 hou rs with no effect by hCG. Interleukin-1 beta significantly increased g ranulosa-luteal cell mRNA levels of IL-1 beta (over twofold), IL-1 rec eptor type 1 (over twofold), PA inhibitor-1 (approximately 1.4-fold), and PA inhibitor-2 (approximately 1.6-fold). In contrast, IL-1 beta ha d no effect on IL-1 beta and IL-1 receptor type 1 mRNA levels in macro phage-depleted granulosa-luteal cells. Granulosa-luteal cells, not mac rophages, account for the majority of the immunocytochemical staining for IL-1 beta and IL-1 receptor type 1 in follicular aspirates. Conclu sions: These studies suggest that the IL-1 system is regulated in huma n granulosa-luteal cells during the periovulatory period. Furthermore, the augmentation of PA inhibitor-1 and -2 mRNA levels by IL-1 beta su ggests a potential role for IL-1 beta in remodeling of the human ovary during the periovulatory period.