Activation of hexosamine pathway impairs nitric oxide (NO)-dependent arteriolar dilations by increased protein O-GlcNAcylation

作者：Beleznai Timea; Bagi Zsolt^*

来源：Vascular Pharmacology, 2012, 56(3-4): 115-121.

DOI：10.1016/j.vph.2011.11.003

• Hyperglycemia
• Glucosamine
• Hexosamine pathway
• Diabetes mellitus
• O-GlcNAcylation
• Microvessel

We hypothesized that under high glucose conditions, activation of the hexosamine pathway leads to impaired nitric oxide (NO)-dependent arteriolar dilation. Skeletal muscle arterioles (diameter: similar to 160 mu m) isolated from male Wistar rats were exposed to normal glucose (NG, 5.5 mmol/L) or high glucose concentrations (HG, 30 mmol/L, for 2 h) and agonist-induced diameter changes were measured with videomicroscopy. Western blots were performed to identify the vascular levels of protein O-linked-N-acetyl-glucosamine (O-GlcNAc) and phosphorylated endothelial NO synthase (eNOS). In arterioles exposed to HG, dilations to histamine were abolished compared to those exposed to NG (max: -6 +/- 6% and 69 +/- 9%, respectively), while acetylcholine-induced responses were not affected. Inhibition of NO synthesis with N-G-nitro-L-arginine methyl ester (L-NAME) reduced histamine-induced dilations in NG arterioles, but it had no effect on microvessels exposed to HG. Dilations to the NO donor, sodium nitroprusside and constrictions to norepinephrine and serotonin were similar in the two groups. In the presence of the inhibitor of hexosamine pathway, azaserine, histamine-induced dilations were significantly augmented in arterioles exposed to HG (max: 67 +/- 2%). Moreover, exposure of vessels to glucosamine (5 mmol/L, for 2 h) resulted in reduced histamine-induced arteriolar dilations (max: 26 +/- 3%). The level of protein O-GlcNAcylation was increased, whereas the P-eNOS (Ser-1177) was decreased in HG exposed vessels. These findings indicate that a high concentration of glucose may lead to glucosamine formation, which impairs histamine-induced, NO-mediated arteriolar dilations. We propose that interfering with the hexosamine pathway may prevent microvascular complications in diabetes.

• 出版日期2012-4