Gper1-but not Esr1 nor Esr2- activation is essential for acute E2-mediated heart protection from ischemic/reperfusion (I/R) injury.

<p>A. Recordings of left ventricular developed pressure (LVDP) comparing heart activity from wild-type (WT) animals not subjected to ischemia/reperfusion (Sham) with the function of hearts subjected to the ischemia reperfusion (I/R) protocol under the continuous presence of vehicle (control) or 40 nM E2 in KH buffer. In sham hearts, function stabilizes at about 30-40 min after mounting and is stable for at least 2 hours. In hearts subjected to I/R (middle and bottom traces), E2 improved recovery from ischemia (during reperfusion) compared to control. B-D. LVDP traces from hearts of Esr1, Esr2 and Gper1 knockouts (-/-) subjected to I/R. Only in Gper1-/- hearts, E2 lost its ability to improve functional recovery (E). n=6-8 hearts/ group.</p

A novel estrogen receptor GPER inhibits mitochondria permeability transition pore opening and protects the heart against ischemia-reperfusion injury

Several studies have recently demonstrated that G protein-coupled receptor 30 (GPER) can directly bind to estrogen and mediate its action. We investigated the role and the mechanism of estrogen-induced cardioprotection after ischemia-reperfusion using a specific GPER agonist G1. Isolated hearts from male mice were perfused using Langendorff technique with oxygenated (95% O2 and 5% CO2) Krebs Henseleit buffer (control), with G1 (1 μM), and G1 (1 μM) together with extracellular signal-regulated kinase (Erk) inhibitor PD-98059 (5μM). After 20 min of perfusion, hearts were subjected to 20 min global normothermic (37°C) ischemia followed by 40 min reperfusion. Cardiac function was measured, and myocardial necrosis was evaluated by triphenyltetrazolium chloride staining at the end of the reperfusion. Mitochondria were isolated after 10 min of reperfusion to assess the Ca2+ load required to induce mitochondria permeability transition pore (mPTP) opening. G1-treated hearts developed better functional recovery with higher rate pressure product (RPP, 6140 ± 264 vs. 2,640 ± 334 beats·mmHg−1·min−1, P < 0.05). The infarct size decreased significantly in G1-treated hearts (21 ± 2 vs. 46 ± 3%, P < 0.001), and the Ca2+ load required to induce mPTP opening increased (2.4 ± 0.06 vs. 1.6 ± 0.11 μM/mg mitochondrial protein, P < 0.05) compared with the controls. The protective effect of G1 was abolished in the presence of PD-98059 [RPP: 4,120 ± 46 beats·mmHg−1·min−1, infarct size: 53 ± 2%, and Ca2+ retention capacity: 1.4 ± 0.11 μM/mg mitochondrial protein (P < 0.05)]. These results suggest that GPER activation provides a cardioprotective effect after ischemia-reperfusion by inhibiting the mPTP opening, and this effect is mediated by the Erk pathway

G Protein-Coupled Estrogen Receptor 1 Mediates Acute Estrogen-Induced Cardioprotection via MEK/ERK/GSK-3β Pathway after Ischemia/Reperfusion.

Three types of estrogen receptors (ER) exist in the heart, Esr1, Esr2 and the G protein-coupled estrogen receptor 1, Gper1. However, their relative importance in mediating estrogen protective action is unknown. We found that, in the male mouse ventricle, Gper1 transcripts are three- and seventeen-fold more abundant than Esr1 and Esr2 mRNAs, respectively. Analysis of the three ER knockouts (Esr1-/-, Esr2-/- and Gper1-/-) showed that only the Gper1-/- hearts lost their ability to be protected by 40 nM estrogen as measured by heart function, infarct size and mitochondrial Ca2+ overload, an index of mitochondrial permeability transition pore (mPTP) activity. Analysis of Akt, ERK1/2 and GSK-3β salvage kinases uncovered Akt and ERK1/2 transient activation by estrogen whose phosphorylation increased during the first 5 min of non-ischemic perfusion. All these increase in phosphorylation effects were abrogated in Gper1-/-. Inhibition of MEK1/2/ERK1/2 (1 μM U0126) and PI-3K/Akt (10 μM LY294002) signaling showed that the MEK1/2/ERK1/2 pathway via GSK-3β exclusively was responsible for cardioprotection as an addition of U0126 prevented estrogen-induced GSK-3β increased phosphorylation, resistance to mitochondrial Ca2+-overload, functional recovery and protection against infarction. Further, inhibiting PKC translocation (1 μM chelerythrin-chloride) abolished estrogen-induced cardioprotection. These data indicate that estrogen-Gper1 acute coupling plays a key role in cardioprotection against ischemia/reperfusion injury in male mouse via a cascade involving PKC translocation, ERK1/2/GSK-3β phosphorylation leading to the inhibition of the mPTP opening