Effect of apocynin on NADPH oxidase-mediated oxidative stress-LOX-1-eNOS pathway in human endothelial cells exposed to high glucose.

Hyperglycemia-induced generation of reactive oxygen species contributes to the development of proatherogenic changes and vasculopathy in diabetes. NADPH oxidase has been recognized as a major source of reactive oxygen species in the vasculature and the lectin-like oxLDL receptor-1 (LOX-1) appears to play a crucial role in the pathogenesis of diabetic endothelial dysfunction. The present study aimed to examine the relationships between the hyperglycemia-mediated NADPH oxidase-LOX-1 pathway activation and nitric oxide-mediated endothelial function. In addition, we investigated effect of the NADPH oxidase inhibitor, apocynin on these consequences. In human umbilical artery endothelial cells (HUAECs), the effect of high glucose on expressional regulations and functional consequences of NADPH oxidase subunits, LOX-1 and endothelial nitric oxide synthase (eNOS), in the absence and presence of apocynin (10 micromol/l) were evaluated. HUAECs were cultured under normal (5.5 mmol/l) or high glucose (30mmol/l) concentrations for 48 h in the absence and presence of apocynin. Our results showed that high glucose significantly enhanced the activity and the protein expression of NADPH oxidase subunits, Nox2 and p47(phox). High glucose markedly increased LOX-1 mRNA level and this was functionally reflected on the augmented uptake of Dil-labelled LDL (5 micromol/l, 3h) by HUAECs. Furthermore, high glucose attenuated eNOS protein and total nitrite levels. However, apocynin inhibited all these changes. Collectively, our study demonstrates that high glucose-induced oxidative stress via NADPH oxidase activation and this contributed to LOX-1 upregulation and eNOS downregulation in human endothelial cells. Apocynin efficiently reversed these consequences, suggesting its potential role as a vasculoprotective agent.

Taye A ，Saad AH ，Kumar AH ，Morawietz H ... -  《-》

High glucose increases expression of cyclooxygenase-2, increases oxidative stress and decreases the generation of nitric oxide in mouse microvessel endothelial cells.

Aljofan M ，Ding H 《-》

Role of LOX-1 in monocyte adhesion-triggered redox, Akt/eNOS and Ca2+ signaling pathways in endothelial cells.

Sakamoto N ，Ishibashi T ，Sugimoto K ，Sawamura T ，Sakamoto T ，Inoue N ，Saitoh S ，Kamioka M ，Uekita H ，Ohkawara H ，Suzuki K ，Teramoto T ，Maruyama Y ，Takeishi Y ... -  《-》

Blockade of chronic high glucose-induced endothelial apoptosis by Sasa borealis bamboo extract.

Choi YJ ，Lim HS ，Choi JS ，Shin SY ，Bae JY ，Kang SW ，Kang IJ ，Kang YH ... -  《-》

Increased oxidative stress in the streptozotocin-induced diabetic apoE-deficient mouse: changes in expression of NADPH oxidase subunits and eNOS.

Ding H ，Hashem M ，Triggle C 《EUROPEAN JOURNAL OF PHARMACOLOGY》