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

Published:November 20, 2013DOI:https://doi.org/10.1016/j.ajog.2013.11.027

      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.

      Key words

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      References

        • Holst R.M.
        • Mattsby-Baltzer I.
        • Wennerholm U.B.
        • Hagberg H.
        • Jacobsson B.
        Interleukin-6 and interleukin-8 in cervical fluid in a population of Swedish women in preterm labor: relationship to microbial invasion of the amniotic fluid, intra-amniotic inflammation, and preterm delivery.
        Acta Obstet Gynecol Scand. 2005; 84: 551-557
        • Martin P.
        • Leibovich S.J.
        Inflammatory cells during wound repair: the good, the bad and the ugly.
        Trends Cell Biol. 2005; 15: 599-607
        • Ferrero-Milani L.
        • Nielsen O.H.
        • Andersen P.S.
        • Girardin S.E.
        Chronic inflammation: importance of NOD2 and NALP3 in interleukin-1beta generation.
        Clin Exp Immunol. 2007; 147: 227-235
        • Marchant D.J.
        • Boyd J.H.
        • Lin D.C.
        • Granville D.J.
        • Garmaroudi F.S.
        • McManus B.M.
        Inflammation in myocardial diseases.
        Circ Res. 2012; 110: 126-144
        • Hansen W.R.
        • Keelan J.A.
        • Skinner S.J.
        • Mitchell M.D.
        Key enzymes of prostaglandin biosynthesis and metabolism: coordinate regulation of expression by cytokines in gestational tissues: a review.
        Prostaglandins Other Lipid Mediat. 1999; 57: 243-257
        • Rauk P.N.
        • Chiao J.P.
        Interleukin-1 stimulates human uterine prostaglandin production through induction of cyclooxygenase-2 expression.
        Am J Reprod Immunol. 2000; 43: 152-159
        • Olson D.M.
        The role of prostaglandins in the initiation of parturition.
        Best Pract Res Clin Obstet Gynaecol. 2003; 17: 717-730
        • Goldenberg R.L.
        • Hauth J.C.
        • Andrews W.C.
        Intrauterine infection and preterm delivery.
        N Engl J Med. 2000; 342: 1500-1507
        • Hedwig S.M.
        Inflammation.
        in: Rubin E. Reisner H.M. Essentials of Rubin's pathology. 5th ed. Lippincott Williams & Wilkins, Philadelphia2009: 23-45
        • Thota C.
        • Farmer T.
        • Garfield R.E.
        • Menon R.
        • Al-Hendy A.
        Vitamin D elicits anti-inflammatory response, inhibits contractile-associated proteins, and modulates Toll-like receptors in human myometrial cells.
        Reprod Sci. 2013; 20: 463-475
        • Aisemberg J.
        • Vercelli C.
        • Billi S.
        • et al.
        Nitric oxide mediates prostaglandins' deleterious effect on lipopolysaccharide-triggered murine fetal resorption.
        Proc Natl Acad Sci. 2007; 104: 7534-7539
        • Pollard J.K.
        • Mitchell M.D.
        Intrauterine infection and the effects of inflammatory mediators on prostaglandin production by myometrial cells from pregnant women.
        Am J Obstet Gynecol. 1996; 174: 682-686
        • Elovitz M.A.
        • Wang Z.
        • Chien E.K.
        • Rychlik D.F.
        • Phillippe M.A.
        New model for inflammation-induced preterm birth: the role of platelet-activating factor and Toll-like receptor-4.
        Am J Pathol. 2003; 163: 2103-2111
        • Helliwell R.J.
        • Keelan J.A.
        • Marvin K.W.
        • et al.
        Gestational age-dependent up-regulation of prostaglandin D synthase (PGDS) and production of PGDS-derived anti-inflammatory prostaglandins in human placenta.
        J Clin Endocrinol Metab. 2006; 91: 597-606
        • Grigsby P.L.
        • Sooranna S.R.
        • Adu-Amankwa B.
        • et al.
        Regional expression of prostaglandin E2 and F2alpha receptors in human myometrium, amnion, and choriodecidua with advancing gestation and labor.
        Biol Reprod. 2006; 75: 297-305
        • Garfield R.E.
        • Sims S.
        • Daniel E.E.
        Gap junctions: their presence and necessity in myometrium during parturition.
        Science. 1977; 198: 958-960
        • Pierce B.T.
        • Calhoun B.C.
        • Adolphson K.R.
        • Lau A.F.
        • Pierce L.M.
        Connexin 43 expression in normal versus dysfunctional labor.
        Am J Obstet Gynecol. 2002; 186: 504-511
        • Soloff M.S.
        • Alexandrova M.
        • Fernstrom M.J.
        Oxytocin receptors: triggers for parturition and lactation?.
        Science. 1979; 204: 1313-1315
        • Mitchell B.F.
        • Schmid B.
        Oxytocin and its receptor in the process of parturition.
        J Soc Gynecol Investig. 2001; 8: 122-133
        • Merlino A.
        • Welsh T.
        • Erdonmez T.
        • et al.
        Nuclear progesterone receptor expression in the human fetal membranes and decidua at term before and after labor.
        Reprod Sci. 2009; 16: 357-363
        • Welsh T.
        • Johnson M.
        • Yi L.
        • et al.
        Estrogen receptor (ER) expression and function in the pregnant human myometrium: estradiol via ERα activates ERK1/2 signaling in term myometrium.
        J Endocrinol. 2012; 212: 227-238
        • How H.
        • Huang Z.H.
        • Zuo J.
        • Lei Z.M.
        • Spinnato 2nd, J.A.
        • Rao C.V.
        Myometrial estradiol and progesterone receptor changes in preterm and term pregnancies.
        Obstet Gynecol. 1995; 86: 936-940
        • Tsuchiya Y.
        • Asano T.
        • Nakayama K.
        • Kato Jr., T.
        • Karin M.
        • Kamata H.
        Nuclear IKKbeta is an adaptor protein for IkappaBalpha ubiquitination and degradation in UV-induced NF-kappaB activation.
        Mol Cell. 2010; 39: 570-582
        • Rabehi L.
        • Irinopoulou T.
        • Cholley B.
        • Haeffner-Cavaillon N.
        • Carreno M.-P.
        Gram-positive and Gram-negative bacteria do not trigger monocytic cytokine production through similar intracellular pathways.
        Infect Immun. 2001; 69: 4590-4599
        • Soloff M.S.
        • Cook Jr., D.L.
        • Jeng Y.J.
        • Anderson G.D.
        In situ analysis of interleukin-1-induced transcription of Cox-2 and IL-8 in cultured human myometrial cells.
        Endocrinology. 2004; 145: 1248-1254
        • White J.H.
        Vitamin D signaling, infectious diseases, and regulation of innate immunity.
        Infect Immun. 2008; 76: 3837-3843
        • Gibney K.B.
        • MacGregor L.
        • Leder K.
        • et al.
        Vitamin D deficiency is associated with tuberculosis and latent tuberculosis infection in immigrants from sub-Saharan Africa.
        Clin Infect Dis. 2008; 46: 443-446
        • Campbell G.R.
        • Spector S.A.
        Vitamin D inhibits human immunodeficiency virus type 1 and Mycobacterium tuberculosis infection in macrophages through the induction of autophagy.
        PLoS Pathog. 2012; 8: e1002689
        • Schleithoff S.S.
        • Zittermann A.
        • Tenderich G.
        • Berthold H.K.
        • Stehle P.
        • Koerfer R.
        Vitamin D supplementation improves cytokine profiles in patients with congestive heart failure: a double-blind, randomized, placebo-controlled trial.
        Am J Clin Nutr. 2006; 83: 754-759
        • Cohen-Lahav M.
        • Shany S.
        • Tobvin D.
        • Chaimovitz C.
        • Douvdevani A.
        Vitamin D decreases NFκB activity by increasing IκBα levels.
        Nephrol Dial Transplant. 2006; 21: 889-897
        • Suzuki K.
        • Bose P.
        • Leong-Quong R.
        • Fujita D.J.
        • Riabowol K.
        REAP: A two minute cell fractionation method.
        BMC Research Notes. 2010; 3: 294
        • Schindler R.
        • Mancilla J.
        • Endres S.
        • Ghorban R.
        • Clark S.C.
        • Dinarello C.A.
        Correlations and interactions in the production of interleukin-6 (IL-6), IL-1, and tumor necrosis factor (TNF) in human blood mononuclear cells: IL-6 suppresses IL-1 and TNF.
        Blood. 1990; 75: 40-47
        • Malaponte G.
        • Bevelacqua V.
        • Fatuzzo P.
        • et al.
        IL-1β, TNF-α and IL-6 release from monocytes in haemodialysis patients in relation to dialytic age.
        Nephrol Dial Transplant. 2002; 17: 1964-1970
        • Romero R.
        • Mazor M.
        • Tartakovsky B.
        Systemic administration of interleukin-1 induces preterm parturition in mice.
        Am J Obstet Gynecol. 1991; 165: 969-971
        • Hirsch E.
        • Filipovich Y.
        • Mahendroo M.
        Signaling via the type I IL-1 and TNF receptors is necessary for bacterially induced preterm labor in a murine model.
        American J Obstet Gynecol. 2006; 194: 1334-1340
        • Sadowsky D.W.
        • Adams K.M.
        • Gravett M.G.
        • Witkin S.S.
        • Novy M.J.
        Preterm labor is induced by intra amniotic infusions of interleukin-1β and tumor necrosis alpha but not by interleukin-6 or interleukin-8 in a nonhuman primate model.
        Am J Obstet Gynecol. 2006; 195: 1578-1589
        • Evans K.N.
        • Nguyen L.
        • Chan J.
        • et al.
        Effects of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 on cytokine production by human decidual cells.
        Biol Reprod. 2006; 75: 816-822
        • Soldin O.P.
        • Guo T.
        • Weiderpass E.
        • et al.
        Steroid hormone levels in pregnancy and 1 year postpartum using isotope dilution tandem mass spectrometry.
        Fertil Steril. 2005; 84: 701-710
        • Birbach A.
        • Gold P.
        • Binder B.R.
        • Hofer E.
        • de Martin R.
        • Schmid J.A.
        Signaling molecules of the NF-B pathway shuttle constitutively between cytoplasm and nucleus.
        J Biol Chem. 2002; 277: 10842-10851
        • Yan X.
        • Sun M.
        • Gibb W.
        Localization of nuclear factor-kappa B (NF kappa B) and inhibitory factor-kappa B (I kappa B) in human fetal membranes and decidua at term and preterm delivery.
        Placenta. 2002; 23: 288-293
        • Tattersall M.
        • Engineer N.
        • Khanjani S.
        • et al.
        Pro-labour myometrial gene expression: are preterm labour and term labour the same?.
        Reproduction. 2008; 135: 569-579