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Impact of Progesterone on Molecular Mechanisms of Preterm Premature Rupture of Membranes

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Abstract

The role and mechanisms of progesterone in preterm premature rupture of membranes (PPROM) remains unclear. This study aims to investigate the molecular mechanisms of action of progesterone in pre-labor full-term fetal amniotic membrane cells with and without stimulation by microbial, pro-inflammatory, or thrombogenic agents. Fetal amniotic membranes were collected from 30 women with a normal singleton pregnancy undergoing elective cesarean section at term prior to the onset of labor. The human amniotic epithelial cells isolated were pretreated with and without medroxyprogesterone acetate for 24 h. Then, cells were treated with and without TLR/NLR agonists, pro-inflammatory cytokines, or thrombin for 48 h. Semi-quantitative RT-PCR, Western blot, and caspase-3 activity measurement were performed. Progesterone stimulation decreased the expression of TLR2, TLR5, and Nod2 genes (alone and/or in combination with TLR/NLR agonists) and decreased the expression of IL-1β and IL-8 genes increased by stimulation with specific agonists for TLR2, TLR4, TLR5, Nod1, and Nod2. Moreover, progesterone decreased thrombin-induced IL-8 gene expression. Progesterone also decreased expression of Bax and Bid proteins (pro-apoptotic factors) increased by stimulation with pro-inflammatory cytokines (TNF-α, NGAL, IL-18, and IL-1β) and thrombin. Progesterone stimulation alone as well as co-stimulation with TNF-α, NGAL, IL-18, IL-1β, or thrombin with progesterone either increased, decreased, or did not change the expression of Bcl-2, Bcl-XL, or XIAP genes (anti-apoptotic factors). These data suggest progesterone plays protective roles against PPROM through anti-microbial, anti-inflammatory, and anti-thrombogenic actions on human-term fetal amniotic membrane cells. Progesterone alters pro-inflammatory cytokine- and thrombin-induced apoptosis by controlling the expression of pro-apoptotic and anti-apoptotic factors.

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Funding

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1A2B1004893).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Inha University Hospital, Inha University School of Medicine, Incheon, Republic of Korea

    Banghyun Lee & Sung Ook Hwang

  2. Department of Obstetrics and Gynecology, Tufts University School of Medicine, Boston, MA, USA

    Errol Norwitz

  3. Mother Infant Research Institute, Tufts Medical Center, Boston, MA, USA

    Errol Norwitz

  4. Department of Obstetrics & Gynecology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

    In Sun Hwang & Jae Yeon Woo

  5. Department of Obstetrics & Gynecology, Uijeongbu St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, 271, Cheonbo-Ro, Uijeongbu-si, Gyeonggi-do, Uijeongbu-si, Republic of Korea

    Hee Joong Lee

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Contributions

Conceptualization: B.L., E.N., S.O.H., and H.J.L.; methodology: B.L., E.N., and H.J.L.; formal analysis and investigation: B.L., I.S.H., J.Y.W., and H.J.L.; writing—original draft preparation: B.L.; writing—review and editing: B.L., E.N., I.S.H., J.Y.W., S.O.H., and H.J.L.; funding acquisition: H.J.L.; resources: B.L., and H.J.L.; supervision: S.O.H., and H.J.L

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Correspondence to Hee Joong Lee.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. This study was approved by the Institutional Review Board of The Catholic Medical Center at the Catholic University of Korea (No. UC19TOSI0123).

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ESM 1

Effects of pro-inflammatory cytokines and thrombin on caspase-3 activity in hAECs with and without progesterone A) TNF-α, B) NGAL, C) IL-18, D) IL-1β, and E) thrombin were used as experimental cytokines. The Y axis is expressed as mean ± SD of experiments from six donors. NS, nonsignificant; SD, standard deviation. (PDF 305 kb)

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Lee, B., Norwitz, E., Hwang, I.S. et al. Impact of Progesterone on Molecular Mechanisms of Preterm Premature Rupture of Membranes. Reprod. Sci. 28, 3137–3146 (2021). https://doi.org/10.1007/s43032-021-00646-8

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  • DOI: https://doi.org/10.1007/s43032-021-00646-8

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