Preconditioning by 17β-estradiol in isolated rat heart depends on PI3-K/PKB pathway, PKC, and ROS

Abstract

To study the cell signaling events leading to 17β-estradiol (E2)-induced acute cardioprotection, we subjected isolated rat hearts to three 5-min cycles of 10 μM E2 before 30 min of regional ischemia, followed by 2 h of reperfusion. Protection was judged by changes in infarct size in percentage of risk zone volume. To test the importance of phosphoinositide 3-kinase (PI3-K), protein kinase C (PKC), or reactive oxygen species (ROS) in E2-induced protection, we combined wortmannin (1 μM), chelerythrine (2 μM), and 2-mercaptopropionylglycine (300 μM), respectively, with E2 exposure. Changes in phosphorylation of protein kinase B (PKB) and selected PKC isoforms were tested by immunoblotting of total lysates and subcellular fractions, along with assessment of PKC translocation from soluble to membrane fraction of heart tissue homogenates. Intracellular ROS levels induced by E2 preconditioning were investigated. E2 preconditioning led to significant reduction in infarct size from 31.8 ± 5.3 to 20.2 ± 2.6% in male hearts and from 42.7 ± 4.7 to 17.1 ± 3.4% in female hearts ( P < 0.05). Protection was abolished by wortmannin (30.0 ± 3.2%), chelerythrine (45.1 ± 4.4%), and 2-mercaptopropionylglycine (36.8 ± 4.7%). E2 preconditioning induced phosphorylation of PKB, PKCα, and PKCε and membrane translocation of PKCε and PKCδ. Intracellular ROS levels were found elevated after transient treatment with hormone. Therefore, our data demonstrate the ability of E2 to induce preconditioning-like cardioprotection via cell signaling events shared by classic ischemic preconditioning.