Vitamin D regulates contractile profile in human uterine myometrial cells via NF-κB pathway

Am J Obstet Gynecol. 2014 Apr;210(4):347.e1-347.e10. doi: 10.1016/j.ajog.2013.11.027. Epub 2013 Nov 18.

Abstract

Objective: Infection triggers inflammation that, in turn, enhances the expression of contractile-associated factors in myometrium and increases the risk of preterm delivery. In this study, we assessed vitamin D regulation of inflammatory markers, contractile-associated factors, steroid hormone receptors, and NFκB pathway proteins in human uterine myometrial smooth muscle (UtSM) cells that were cultured in an inflammatory environment.

Study design: Inflammatory environment was simulated for UtSM cells by coculturing them with monocyte lineage (THP1) cells. We measured the expression of inflammatory markers, contractile-associated factors, steroid hormone receptors, and NFκB pathway proteins in UtSM cells that were cultured with THP1 cells in the presence and absence of vitamin D by real time polymerase chain reaction and Western blot analysis.

Results: Monocytes secreted monocyte inflammatory protein-1α and -1β, interleukin (IL)-1β and 6, and tumor necrosis factor-α into the conditioned medium. In the UtSM cells that had been cocultured with THP1 cells, there was a significant (P < .05) increase in the expression of inflammatory markers IL-1β, -6, and -13 and tumor necrosis factor-α; the contractile-associated factors connexin-43, Cox-2, and prostaglandin F2α receptor; the estrogen receptor α, and progesterone receptors A and B. Vitamin D treatment of cocultures decreased (P < .05) the expression of inflammatory markers and contractile-associated factors in UtSM cells. Similarly, vitamin D decreased estrogen receptor α and progesterone receptors A-to-B ratio in UtSM cells that were cocultured with THP1 cells. In addition, vitamin D treatment significantly (P < .05) decreased monocyte-induced p-IκBα in cytosol and NFκB-p65 in the nucleus and increased IκBα in cytosol in UtSM cells.

Conclusion: Our results suggest that vitamin D treatment decreases inflammation-induced cytokines and contractile-associated factors in the uterine myometrial smooth muscle cells through the NFκB pathway.

Keywords: NFκB; contractile-associated factor; inflammatory marker; monocyte; myometrial cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Anti-Inflammatory Agents / pharmacology*
  • Blotting, Western
  • Cell Line
  • Coculture Techniques
  • Connexin 43 / genetics
  • Connexin 43 / metabolism
  • Cyclooxygenase 2 / metabolism
  • Cytokines / genetics
  • Cytokines / metabolism
  • Cytosol / metabolism
  • Estrogen Receptor alpha / metabolism
  • Female
  • Humans
  • I-kappa B Proteins / metabolism
  • Muscle Contraction / drug effects*
  • Myometrium / cytology*
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / metabolism*
  • Phosphorylation
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Receptors, Oxytocin / genetics
  • Receptors, Oxytocin / metabolism
  • Receptors, Progesterone / metabolism
  • Receptors, Prostaglandin / genetics
  • Receptors, Prostaglandin / metabolism
  • Vitamin D / pharmacology*

Substances

  • Anti-Inflammatory Agents
  • Connexin 43
  • Cytokines
  • Estrogen Receptor alpha
  • GJA1 protein, human
  • I-kappa B Proteins
  • NF-kappa B
  • NFKBIA protein, human
  • RNA, Messenger
  • Receptors, Oxytocin
  • Receptors, Progesterone
  • Receptors, Prostaglandin
  • progesterone receptor A
  • progesterone receptor B
  • prostaglandin F2alpha receptor
  • NF-KappaB Inhibitor alpha
  • Vitamin D
  • Cyclooxygenase 2
  • PTGS2 protein, human