rage modulates hypoxiareoxygenation injury in adult murine cardiomyocytes via jnk and gsk-3β signaling pathways愤怒在成年小鼠心肌细胞通过调节hypoxiareoxygenation损伤物和gsk-3β信号通路.pdfVIP

RAGE Modulates Hypoxia/Reoxygenation Injury in Adult Murine Cardiomyocytes via JNK and GSK-3b Signaling Pathways Linshan Shang, Radha Ananthakrishnan, Qing Li, Nosirudeen Quadri, Mariane Abdillahi, Zhengbin Zhu, ´ Wu Qu, Rosa Rosario, Fatouma Toure, Shi Fang Yan, Ann Marie Schmidt, Ravichandran Ramasamy* Division of Surgical Science, Department of Surgery, College of Physicians and Surgeons, Columbia University, New York, New York, United States of America Abstract Background: Advanced 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 Findings: Cardiomyocytes 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.