Androgen-Induced Relaxation of Uterine Myocytes Is Mediated by Blockade of Both Ca2+ Flux and MLC Phosphorylation

• Published in: The journal of clinical endocrinology and metabolism 2016-03, Vol.101 (3), p.1055-1065
• Main Authors: Makieva, Sofia, Hutchinson, Lawrence J, Rajagopal, Shalini P, Rinaldi, Sara F, Brown, Pamela, Saunders, Philippa T. K, Norman, Jane E
• Format: Article
• Language: English
• ISSN: 0021-972X; 1945-7197
• DOI: 10.1210/jc.2015-2851

Context: Uterine quiescence must be maintained until pregnancy reaches term. Premature activation of myometrial contractility leads to preterm labor and delivery. Objective: To scrutinize the potential of androgens to relax the myometrium and the mechanism of their action. Samples: A pregnancy-derived myometrial smooth muscle cell line (PHM1-41) and myometrial strips prepared from tissues obtained from pregnant women (lean, n = 9; obese, n = 6) undergoing elective cesarean section at term and from nonpregnant C57BL/6 mice (n=5) were each utilized. Design: The contraction of collagen-embedded PHM1–41s and the stretch-induced contraction of human and murine myometrial strips were assessed after incubation with Testosterone (T), dihydrotestosterone (DHT), and T conjugated to BSA. Intracellular calcium ([Ca2+]) and phosphorylated myosin light chain concentrations were quantified in PHM1–41s using a Fluo-4 Ca2+ assay and in-cell Westerns, respectively. Setting: University research institute. Results: DHT and T, but not T conjugated to BSA, impaired the contractile function of PHM1–41s and of human and murine myometrial strips. The response was rapid (observed within minutes), was sustainable for up to 48 hours, and was not abolished on knockdown of the androgen receptor. DHT (100 μm) reduced the amplitude of lean strip contraction to 2 ± 2% of the pretreatment value and T (100 μm) to 3.3 ± 1%. These values for obese strips were 15 ± 6.7% and 11± 6.7%, respectively. At the same doses, in murine strips, DHT reduced the amplitude to 4.8 ± 3% and T to 4.9 ± 3%. DHT (50 μm) pretreatment reduced the oxytocin-stimulated increase in [Ca2+] (P < .0001; n = 6) and phosphorylated myosin light chain (P < .05; n = 5) in PHM1–41s. Conclusion: Lipid-soluble androgens could be developed as tocolytic agents for the treatment of preterm labor.