Abstract
BackgroundAdvanced glycation end-products (AGEs) have been implicated in diverse pathological settings including diabetes, inflammation and acute ischemia/reperfusion injury in the heart. AGEs interact with the receptor for AGEs (RAGE) and transduce signals through activation of MAPKs and proapoptotic pathways. In the current study, adult cardiomyocytes were studied in an in vitro ischemia/reperfusion (I/R) injury model to delineate the molecular mechanisms underlying RAGE-mediated injury due to hypoxia/reoxygenation (H/R).Methodology/Principal FindingsCardiomyocytes isolated from adult wild-type (WT), homozygous RAGE-null (RKO), and WT mice treated with soluble RAGE (sRAGE) were subjected to hypoxia for 30 minutes alone or followed by reoxygenation for 1 hour. In specific experiments, RAGE ligand carboxymethyllysine (CML)-AGE (termed “CML” in this manuscript) was evaluated in vitro. LDH, a marker of cellular injury, was assayed in the supernatant in the presence or absence of signaling inhibitor-treated cardiomyocytes. Cardiomyocyte levels of heterogeneous AGEs were measured using ELISA. A pronounced increase in RAGE expression along with AGEs was observed in H/R vs. normoxia in WT cardiomyocytes. WT cardiomyocytes after H/R displayed increased LDH release compared to RKO or sRAGE-treated cardiomyocytes. Our results revealed significant increases in phospho-JNK in WT cardiomyocytes after H/R. In contrast, neither RKO nor sRAGE-treated cardiomyocytes exhibited increased phosphorylation of JNK after H/R stress. The impact of RAGE deletion on GSK-3β phosphorylation in the cardiomyocytes subjected to H/R revealed significantly higher levels of phospho-GSK-3β/total GSK-3β in RKO, as well as in sRAGE-treated cardiomyocytes versus WT cardiomyocytes after H/R. Further investigation established a key role for Akt, which functions upstream of GSK-3β, in modulating H/R injury in adult cardiomyocytes.Conclusions/SignificanceThese data illustrate key roles for RAGE-ligand interaction in the pathogenesis of cardiomyocyte injury induced by hypoxia/reoxygenation and indicate that the effects of RAGE are mediated by JNK activation and dephosphorylation of GSK-3β. The outcome in this study lends further support to the potential use of RAGE blockade as an adjunctive therapy for protection of the ischemic heart.
Highlights
Mounting evidence indicates that ischemia and ischemia/ reperfusion (I/R) play important roles in cardiomyocyte loss in pathophysiological conditions
Thirty minutes of hypoxia followed by 1 hr of reoxygenation resulted in
As H/R resulted in increased expression of RAGE in cardiomyocytes, we sought to identify if H/R resulted in increased generation of RAGE ligand Advanced glycation end-products (AGEs)
Summary
Mounting evidence indicates that ischemia and ischemia/ reperfusion (I/R) play important roles in cardiomyocyte loss in pathophysiological conditions. The myocyte may be exposed to a variety of cellular stresses, such as hypoxia, ischemia, and I/R Cardiomyocyte loss, both necrotic and apoptotic, is a feature of many pathological conditions in the heart [1]. AGEs interact with the receptor for AGEs (RAGE) which results in the propagation of stress signals and activation of MAPKs, NFkB, and several proapoptotic pathways [8,9]. Adult cardiomyocytes were used as an in vitro I/R injury model to delineate the molecular mechanisms by which RAGE mediates injury due to hypoxia and reoxygenation. Adult cardiomyocytes were studied in an in vitro ischemia/reperfusion (I/R) injury model to delineate the molecular mechanisms underlying RAGEmediated injury due to hypoxia/reoxygenation (H/R)
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have