Anti-inflammatory effect of erythropoietin pretreatment on cardiomyocytes with hypoxia/reoxygenation injury and the possible mechanism

• Published in: Chinese journal of traumatology Vol. 11; no. 6; pp. 352 - 358
• Main Author: 秦川 肖颖彬 钟前进 陈林 王学锋
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.12.2008; Department of Cardiovascular Surgery, Xinqiao Hospital,Third Military Medical University, Chongqing 400037, China
• Subjects: Analysis of Variance; Animals; Animals, Newborn; Antioxidants - pharmacology; Blotting, Western; Cells, Cultured; Electrophoretic Mobility Shift Assay; Erythropoietin; Erythropoietin - pharmacology; Hypoxia - metabolism; Inflammation - drug therapy; Myocytes, cardiac; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; NF-kappa B - biosynthesis; Pyrrolidines - pharmacology; Random Allocation; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Reperfusion Injury - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Thiocarbamates - pharmacology; Tumor necrosis factor-alpha; Tumor Necrosis Factor-alpha - antagonists & inhibitors; Tumor Necrosis Factor-alpha - biosynthesis; Tumor Necrosis Factor-alpha - genetics; 强心剂; 红细胞生成素; 肌细胞; Erythropoietin; Tumor necrosis factor-alpha; Myocytes, cardiac; Myocytes,cardiac
• ISSN: 1008-1275
• DOI: 10.1016/S1008-1275(08)60071-1

Objective： To investigate the anti-inflammatory effect of erythropoietin （EPO） pretreatment on cardiomyocytes exposed to hypoxialreoxygenation injury （H/R） and explore the possible mechanism. Methods： The cultured neonatal rats＇ ventricular cardiomyocytes were divided randomly into 4 groups, control group （C group）, EPO pretreatment group （E group）, EPO and pyrrolidine dithiocarbamate （PDTC） pretreatment group （EP group） and PDTC pretreatment group （P group）. After 24 hours＇ pretreatment, the cardiomyocytes were exposed to H/R. After pretreatment and H/R, the expression of tumor necrosis factor- α （TNF- α ） gene in all the groups was detected by RT-PCR and Western blot. The nuclear factor- κ B （NF- κB） activity was detected by electrophoretic mobility shift assay （EMSA） and the inhibitor- κB α （Ⅰ- κB α） protein level was detected by Western blot. Results： The decrement of Ⅰ- κB a protein and the increasing NF- KB activity were found in cardiomyocytes pretreated with EPO before H/R compared to other groups （t=3.321, 4.183, P〈0.01）. However, after H/R, NF- κB activity and expression of TNF- α gene were significantly reduced, Ⅰ- κB a protein expression was increased in cardiomyocytes of E group compared to other groups （t=-3.425, 3.687, 3.454, P〈0.01）. All theses changes caused by EPO pretreatment were eliminated by the intervention of PDTC （an antagonist to NF- κB） during pretreatment. Conclusions： EPO pretreatment can inhibit the activation of NF- κB and upregulation of TNF- α gene in cardiomyocytes exposed to H/R through a negative feedback of NF- κB signaling pathway, and thus produces the anti-inflammatory effect. This might be one of the ways EPO produces the anti-inflammatory effect.