Cholinergic drugs ameliorate endothelial dysfunction by decreasing O-GlcNAcylation via M3 AChR-AMPK-ER stress signaling

• Published in: Life sciences (1973) 2019-04, Vol.222, p.1-12
• Main Authors: Cui, Yan-Ling, Xue, Run-Qing, Xi He, Ming Zhao, Yu, Xiao-Jiang, Liu, Long-Zhu, Wu, Qing, Si Yang, Li, Dong-Ling, Zang, Wei-Jin
• Format: Article
• Language: English
• Subjects: Acetylcholine; Acetylcholine receptors (muscarinic); AMP; AMP-activated protein kinase; Apoptosis; Body weight; Butyric acid; Cardiovascular diseases; Cholinergic drugs; Cholinergics; Endoplasmic reticulum; Endothelial cell apoptosis; Endothelial cells; Endothelium; ER stress; Glucose; High fat diet; Inhibitors; Kinases; M3 AChR-AMPK; Mice; Morbidity; N-Acetylglucosamine; O-GlcNAc; O-GlcNAcylation; Obesity; Proteins; Pyridostigmine; Ribose; Signaling; Stress; Time dependence; Umbilical vein; Weight reduction
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2019.02.036

Obesity is associated with increased cardiovascular morbidity and mortality. It is accompanied by augmented O-linked β-N-acetylglucosamine (O-GlcNAc) modification of proteins via increasing hexosamine biosynthetic pathway (HBP) flux. However, the changes and regulation of the O-GlcNAc levels induced by obesity are unclear. High fat diet (HFD) model was induced obesity in mice with or without the cholinergic drug pyridostigmine (PYR, 3 mg/kg/d) for 22 weeks and in vitro human umbilical vein endothelial cells (HUVECs) was treated with high glucose (HG, 30 mM) with or without acetylcholine (ACh). PYR significantly reduced body weight, blood glucose, and O-GlcNAcylation levels and attenuated vascular endothelial cells detachment in HFD-fed mice. HG addition induced endoplasmic reticulum (ER) stress and increased O-GlcNAcylation levels and apoptosis in HUVECs in a time-dependent manner. Additionally, HG decreased levels of phosphorylated AMP-activated protein kinase (AMPK). Interestingly, ACh significantly blocked damage to HUVECs induced by HG. Furthermore, the effects of ACh on HG-induced ER stress, O-GlcNAcylation, and apoptosis were prevented by treating HUVECs with 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP, a selective M3 AChR antagonist) or compound C (Comp C, an AMPK inhibitor). Treatment with 5-aminoimidazole-4-carboxamide ribose (AICAR, an AMPK activator), 4-phenyl butyric acid (4-PBA, an ER stress inhibitor), and 6-diazo-5-oxonorleucine (DON, a GFAT antagonist) reproduced a similar effect with ACh. Activation of cholinergic signaling ameliorated endothelium damage, reduced levels of ER stress, O-GlcNAcylation, and apoptosis in mice and HUVECs under obese conditions, which may function through M3 AChR-AMPK signaling.