Naringin, an active flavonoid isolated from citrus fruit extracts, exhibits biological and pharmacological properties, such as antioxidant activity and antidiabetic effect. Mitogen-activated protein kinase (MAPK) signalling pathway has been shown to participate in hyperglycaemia-induced injury. The present study tested the hypothesis that naringin protects against high glucose (HG)-induced injuries by inhibiting MAPK pathway in H9c2 cardiac cells. To examine this, the cells were treated with 35 mM glucose (HG) for 24 hr to establish a HG-induced cardiomyocyte injury model. The cells were pre-treated with 80 lM naringin for 2 hr before exposure to HG. The findings of this study showed that exposure of H9c2 cells to HG for 24 hr markedly induced injuries, as evidenced by a decrease in cell viability, increases in apoptotic cells and reactive oxygen species (ROS) production, as well as dissipation of mitochondrial membrance potential (MMP). These injuries were significantly attenuated by the pre-treatment of cells with either naringin or SB203580 (a selective inhibitor of p38 MAPK) or U0126 (a selective inhibitor of extracellular signal regulated kinase 1/2, ERK1/2) or SP600125 (a selective inhibitor of c-jun N-termanal kinase, JNK) before exposure to HG, respectively. Furthermore, exposure of cells to HG increased the phosphorylation of p38 MAPK, ERK1/2 and JNK. The increased activation of MAPK pathway was ameliorated by pre-treatment with either naringin or N-acetyl-L-cysteine (NAC), a ROS scavenger, which also reduced HG-induced cytotoxicity and apoptosis, leading to increase in cell viability and decrease in apoptotic cells. In conclusion, our findings provide new evidence for the first time that naringin protects against HGinduced injuries by inhibiting the activation of MAPK (p38 MAPK, ERK1/2 and JNK) and oxidative stress in H9c2 cells.Hyperglycaemia, the most important feature of diabetes mellitus (DM), is believed to be a risk factor for the development of diabetic cardiovascular complications, such as diabetic cardiomyopathy (DCM) [1,2]. The mechanisms responsible for the deteriorative effects of hyperglycaemia on cardiomyocytes are complicated. Multiple factors, such as reactive oxygen species (ROS) production [3][4][5][6][7][8], decline of antioxidant defence systems [6-10], pro-inflammatory cytokine [11][12][13], activation of mitogen-activated protein kinase (MAPK) [14][15][16] and mitochondrial damage [17,18] have been reported to contribute to hyperglycaemia-induced injuries. Sustained hyperglycaemia has been identified as a principle mediator of ROS generation in diabetes [3,4,14,19], which leads to oxidative myocardial injury [14,20]. Furthermore, hyperglycaemia significantly increases phosphorylation of p38 MAPK and extracellular signal regulated kinase (ERK) 1/2 (members of MAPK) in left ventricle tissue of diabetic rats [14], the increased activation of MAPK is associated with the development of diabetic cardiomyopathy [14]. In streptozotocin (STZ)-induced diabetic rats, hyperglycaemia...