Involvement of sarco/endoplasmic reticulum Ca2 +-ATPase (SERCA) in mPRα (PAQR7)-mediated progesterone induction of vascular smooth muscle relaxation

Progesterone acts directly on vascular smooth muscle cells (VSMCs) through activation of membrane Progesterone Receptor α (mPRα)-dependent signaling to rapidly decrease cytosolic CA²⁺ concentrations and induce muscle relaxation. However, it is not known whether this progesterone action involves uptake of CA²⁺ by the sarco/endoplasmic reticulum (SR) and increased sarco/endoplasmic reticulum CA²⁺-ATPase (SERCA) activity. The present results show that treatment of cultured human VSMCs with progesterone and the selective mPR agonist Org OD-02-0 (OD 02-0) but not with the nuclear PR agonist R5020 increased SERCA protein expression, which was blocked by knockdown of mPRα with siRNA. Moreover, treatments with progesterone and OD 02-0, but not with R5020, increased phospholamban (PLB) phosphorylation, which would result in disinhibition of SERCA function. Progesterone and OD 02-0 significantly increased CA²⁺ levels in the SR and caused VSMC relaxation. These effects were blocked by pretreatment with cyclopiazonic acid (CPA), a SERCA inhibitor, and by knockdown of SERCA2 with siRNA, suggesting that SERCA2 plays a critical role in progesterone induction of VSMC relaxation. Treatment with inhibitors of inhibitory G proteins (Gi, NF023), MAP kinase (AZD 6244), Akt/PI3K (wortmannin), and a Rho activator (calpeptin) blocked the progesterone- and OD 02-0-induced increase in CA²⁺ levels in the SR and SERCA expressions. These results suggest that the rapid effects of progesterone on cytosolic CA²⁺ levels and relaxation of VSMCs through mPRα involve regulation of the functions of SERCA2 and PLB through Gi, MAP kinase, and Akt signaling pathways and downregulation of RhoA activity.NEW & NOTEWORTHY The rapid effects of progesterone on cytosolic CA²⁺ levels and relaxation of VSMCs through mPRα involve regulation of the functions of SERCA2 and PLB through Gi, MAP kinase, and Akt signaling pathways and downregulation of RhoA activity.