17β-estradiol induces hyperresponsiveness in guinea pig airway smooth muscle by inhibiting the plasma membrane Ca2+-ATPase

• Published in: Molecular and cellular endocrinology 2024-09, Vol.590, p.112273, Article 112273
• Main Authors: Romero-Martínez, Bianca S., Flores-Soto, Edgar, Sommer, Bettina, Reyes-García, Jorge, Arredondo-Zamarripa, David, Solís-Chagoyán, Héctor, Lemini, Cristina, Rivero-Segura, Nadia A., Santiago-de-la- Cruz, José A., Pérez-Plascencia, Carlos, Montaño, Luis M.
• Format: Article
• Language: English
• Subjects: 17β-estradiol; acute exposure; adenosinetriphosphatase; Airway hyperresponsiveness; Airway smooth muscle; asthma; Asthmatic women; blood serum; Ca2-transporting ATPase; caffeine; calcium; carbachol; choline chloride; cyclopiazonic acid; estradiol; guinea pigs; lanthanum; mitochondria; muscle contraction; myocytes; plasma membrane; Plasma membrane Ca2+-ATPase pump; sarcoplasmic reticulum; smooth muscle; Western blotting
• ISSN: 0303-7207; 1872-8057; 1872-8057
• DOI: 10.1016/j.mce.2024.112273

High serum estrogen concentrations are associated with asthma development and severity, suggesting a link between estradiol and airway hyperresponsiveness (AHR). 17β-estradiol (E2) has non-genomic effects via Ca2+ regulatory mechanisms; however, its effect on the plasma membrane Ca2+-ATPases (PMCA1 and 4) and sarcoplasmic reticulum Ca2+-ATPase (SERCA) is unknown. Hence, in the present study, we aim to demonstrate if E2 favors AHR by increasing intracellular Ca2+ concentrations in guinea pig airway smooth muscle (ASM) through a mechanism involving Ca2+-ATPases. In guinea pig ASM, Ca2+ microfluorometry, muscle contraction, and Western blot were evaluated. Then, we performed molecular docking analysis between the estrogens and Ca2+ ATPases. In tracheal rings, E2 produced AHR to carbachol. In guinea pig myocytes, acute exposure to physiological levels of E2 modified the transient Ca2+ peak induced by caffeine to a Ca2+ plateau. The incubation with PMCA inhibitors (lanthanum and carboxyeosin, CE) partially reversed the E2-induced sustained plateau in the caffeine response. In contrast, cyclopiazonic acid (SERCA inhibitor), U-0126 (an inhibitor of ERK 1/2), and choline chloride did not modify the Ca2+ plateau produced by E2. The mitochondrial uniporter activity and the capacitative Ca2+ entry were unaffected by E2. In guinea pig ASM, Western blot analysis demonstrated PMCA1 and PMCA4 expression. The results from the docking modeling demonstrate that E2 binds to both plasma membrane ATPases. In guinea pig tracheal smooth muscle, inhibiting the PMCA with CE, induced hyperresponsiveness to carbachol. 17β-estradiol produces hyperresponsiveness by inhibiting the PMCA in the ASM and could be one of the mechanisms responsible for the increase in asthmatic crisis in women. A) The Caf-induced transient Ca2+ peak is produced by the release of Ca2+ from the SR by the RyR and the decrease of cytosolic Ca2+ by the PMCA and SERCA. B) The inhibition of PMCA by E2 modifies the Ca2+ response to Caf, inducing a Ca2+ plateau. C) this sustained [Ca2+]i leads to airway hyperresponsiveness. PMCA, plasma membrane Ca2+ ATPase; Caf, caffeine; RyR, ryanodine receptor; SERCA, sarcoplasmic reticulum Ca2+ ATPase; SR, sarcoplasmic reticulum; E2, 17β-estradiol; ASM, airway smooth muscle. [Display omitted] •Acute exposure to 17β-estradiol (E2) induces airway hyperresponsiveness (AHR) in the airway smooth muscle (ASM).•The acute incubation of E2 in ASM cells modified the transient Ca2+ peak ind