Macrophage-induced reactive oxygen species promote myometrial contraction and labor-associated mechanisms.

Fiche publication


Date publication

mars 2020

Journal

Biology of reproduction

Auteurs

Membres identifiés du Cancéropôle Est :
Dr GARRIDO Carmen, Pr LIRUSSI Frédéric, Dr WENDREMAIRE Maeva


Tous les auteurs :
Wendremaire M, Hadi T, Pezze M, Barrichon M, Lopez T, Neiers F, Sagot P, Garrido C, Lirussi F

Résumé

At labor, the myometrium is infiltrated by a massive influx of macrophages that secrete high levels of pro-inflammatory cytokines inducing the expression of specific labor-associated markers. However, the interactions between myocytes and macrophages and the role of macrophages in the myometrium at labor remain to be elucidated. In this work, we studied the role of myometrium-infiltrated macrophages and their interaction with myocytes in lipopolysaccharide-induced preterm labor. A co-culture model of human primary myometrial cells and macrophages was developed and validated. Collagen lattices were used to evaluate myocyte contraction. Differentiation steps were assessed by i) phalloidin and vinculin staining for cytoskeleton reorganization, ii) gap junction protein alpha 1 expression and scrape loading/dye transfer with Lucifer Yellow for gap junction intercellular communication, and iii) calcium imaging for cell excitability. We demonstrated that macrophages favored lipopolysaccharide-induced contraction and early differentiation of myometrial cells. Transwell assays showed that previous activation of macrophages by lipopolysaccharide was essential for this differentiation and that macrophage/myocyte interactions involved macrophage release of reactive oxygen species (ROS). The effects of macrophage-released ROS in myometrial cell transactivation were mimicked by H2O2, suggesting that superoxide anion is a major intermediate messenger in macrophage/myocyte crosstalk during labor. These novel findings provide the foundation for innovative approaches to managing preterm labor, specifically the use of antioxidants to inhibit the initial stages of labor before the contractile phenotype has been acquired. In addition, the co-culture model developed by our team could be used in future research to decipher pathophysiological signaling pathways or screen/develop new tocolytics.

Mots clés

Cell culture, Differentiation, Labor, Macrophage, Myocyte, Oxidative stress

Référence

Biol. Reprod.. 2020 Mar 13;: