Abstract.Glucosamine is a possible cause of vascular endothelial injury in the initial stages of atherosclerosis, through endoplasmic reticulum (ER) stress resulting in fatty streaks in the vascular wall. Quercetin is an anti-diabetic and cardiovascular protective agent that has previously been demonstrated to reduce ER stress in human umbilical vein endothelial cells (HUVECs). The present study aimed to investigate whether quercetin prevents glucosamine-induced apoptosis and inflammation via ER stress pathway in HUVECs. The effect of quercetin on cell viability, apoptosis, and protein expression levels of inflammatory cytokines and ER stress markers was investigated in glucosamine-supplemented HUVECs. Quercetin was demonstrated to protect against glucosamine-induced apoptosis, improved cell viability, and inhibited expression of pro-inflammatory factors and endothelin-1. Quercetin treatment also reduced the expression levels of glucose-regulated protein 78, phosphorylated protein kinase-like ER kinase, phosphorylated c-Jun N-terminal kinase and C/EBP homologous protein. In conclusion, quercetin may have auxiliary therapeutic potential against glucosamine-induced cell apoptosis and inflammation, which may be partially due to alleviation of ER stress.
IntroductionDiabetes is a 'coronary heart disease (CHD) equivalent' disease, as epidemiological data has previously demonstrated that patients with diabetes without any prior evidence of CHD are at greater risk of death from CHD than nondiabetic patients with prior evidence of CHD (1,2). Atherosclerosis (AS) is a disease of arterial lipid deposition leading to a number of biological responses, including a chronic, macrophage-dominated inflammatory reaction (3), and is the pathological basis of CHD. The hexosamine biosynthesis pathway (HBP), a normal pathway for glucose metabolism, is activated excessively in patients with diabetes, resulting in increased cellular glucosamine (4). Endoplasmic reticulum (ER) is a membranous network to synthesize, modify, fold and assemble proteins. When the ER function, particularly folding capacity, is challenged, the unfolded protein response (UPR) is executed as a protective mechanism in ER. Failure of this mechanism to fold newly synthesized proteins shows unique damage to the cell and is termed ʻER stressʼ (5,6). Numerous studies have highlighted that ER stress may link hyperglycemia to AS (4,7,8). Important ER stress markers, including protein kinase-like ER kinase (PERK), glucose regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP), were expressed in the arterial wall of streptozotocin-induced hyperglycemic apolipoprotein (apoE)-deficient mice (4). Glucosamine levels and the expression of GRP78 were increased following hyperglycemia and prior to the early stages of fatty streak formation in aortic endothelial cells of hyperglycemic apoE -/-mice (7). In vitro studies have demonstrated that incubation of hepatic cells (9) and adipocytes (10) with 5 mM glucosamine resulted in lipid accumulation, impaired insulin-stimul...