Chelsea W Fox1, Lingzhi Zhang2, Benjamin C Moeller3, V Gabriel Garzo4, R Jeffrey Chang2, Antoni J Duleba2. 1. Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, University of South Carolina School of Medicine/Prisma Health, Greenville, South Carolina. 2. Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, La Jolla, San Diego, California. 3. California Animal Health and Food Safety Laboratory, School of Veterinary Medicine, University of California, Davis, California. 4. Reproductive Partners Fertility Center-San Diego, La Jolla, San Diego, California.
Abstract
OBJECTIVE: To study the effects of ibuprofen on androgen production, gene expression, and cell viability in rat theca-interstitial cells exposed to the proinflammatory stimuli interleukin-1β (IL-1β) and lipopolysaccharide (LPS). DESIGN: Animal study. SETTING: University-based research laboratory. PATIENTS/ANIMALS: Theca-interstitial cells were isolated from 30 day old female Sprague Dawley rats. INTERVENTIONS: Theca cells were cultured with pro-inflammatory media containing IL-1β and LPS and compared with cells cultured in control media. MAIN OUTCOME MEASURES: Androstenedione quantification was performed on conditioned cell culture medium using liquid chromatography-mass spectrometry. Theca cell viability was assessed using PrestoBlue cell viability assay. The gene expression of Cyp17a1, Cyp11a1, and Hsd3b was analyzed using quantitative polymerase chain reaction. RESULTS: Both proinflammatory stimuli IL-1β and LPS increased androstenedione concentration in cell culture medium, and these effects were mitigated with ibuprofen. Both inflammatory agents in addition increased the expression of key genes involved in androgen synthesis: Cyp17a1, Cyp11a1, and Hsd3b; the addition of ibuprofen to the culture medium inhibited these effects. Theca cell viability increased with IL-1β and LPS. Ibuprofen inhibited the IL-1β-mediated increase in cell viability but did not reverse the effects of LPS. CONCLUSIONS: In conclusion, our findings support the hypothesis that many of the alterations induced by inflammatory stimuli in theca-interstitial cells are abrogated by the addition of ibuprofen.
OBJECTIVE: To study the effects of ibuprofen on androgen production, gene expression, and cell viability in rat theca-interstitial cells exposed to the proinflammatory stimuli interleukin-1β (IL-1β) and lipopolysaccharide (LPS). DESIGN: Animal study. SETTING: University-based research laboratory. PATIENTS/ANIMALS: Theca-interstitial cells were isolated from 30 day old female Sprague Dawley rats. INTERVENTIONS: Theca cells were cultured with pro-inflammatory media containing IL-1β and LPS and compared with cells cultured in control media. MAIN OUTCOME MEASURES: Androstenedione quantification was performed on conditioned cell culture medium using liquid chromatography-mass spectrometry. Theca cell viability was assessed using PrestoBlue cell viability assay. The gene expression of Cyp17a1, Cyp11a1, and Hsd3b was analyzed using quantitative polymerase chain reaction. RESULTS: Both proinflammatory stimuli IL-1β and LPS increased androstenedione concentration in cell culture medium, and these effects were mitigated with ibuprofen. Both inflammatory agents in addition increased the expression of key genes involved in androgen synthesis: Cyp17a1, Cyp11a1, and Hsd3b; the addition of ibuprofen to the culture medium inhibited these effects. Theca cell viability increased with IL-1β and LPS. Ibuprofen inhibited the IL-1β-mediated increase in cell viability but did not reverse the effects of LPS. CONCLUSIONS: In conclusion, our findings support the hypothesis that many of the alterations induced by inflammatory stimuli in theca-interstitial cells are abrogated by the addition of ibuprofen.
Authors: Chelsea W Fox; Lingzhi Zhang; Abhishek Sohni; Manuel Doblado; Miles F Wilkinson; R Jeffrey Chang; Antoni J Duleba Journal: Endocrinology Date: 2019-12-01 Impact factor: 4.736
Authors: Israel Ortega; Jesus A Villanueva; Donna H Wong; Amanda B Cress; Anna Sokalska; Scott D Stanley; Antoni J Duleba Journal: J Ovarian Res Date: 2014-02-13 Impact factor: 4.234