20-Hydroxyeicosatetraenoic acid impairs endothelial insulin signaling by inducing phosphorylation of the insulin receptor substrate-1 at Ser616

Xuguang Li; Gang Zhao; Ben Ma; Rui Li; Jiang Hong; Shaowen Liu; Dao Wen Wang

doi:10.1371/journal.pone.0095841

20-Hydroxyeicosatetraenoic acid impairs endothelial insulin signaling by inducing phosphorylation of the insulin receptor substrate-1 at Ser616

PLoS One. 2014 Apr 24;9(4):e95841. doi: 10.1371/journal.pone.0095841. eCollection 2014.

Authors

Xuguang Li¹, Gang Zhao², Ben Ma², Rui Li², Jiang Hong¹, Shaowen Liu¹, Dao Wen Wang²

Affiliations

¹ Department of Cardiology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
² Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

Abstract

20-Hydroxyeicosatetraenoic acid (20-HETE) induces endothelial dysfunction and is correlated with diabetes. This study was designed to investigate the effects of 20-HETE on endothelial insulin signaling.Human umbilical vein endothelial cells (HUVECs) or C57BL/6J mice were treated with 20-HETE in the presence or absence of insulin, and p-ERK1/2, p-JNK, IRS-1/PI3K/AKT/eNOS pathway, were examined in endothelial cells and aortas by immunoblotting. eNOS activity and nitric oxide production were measured. 20-HETE increased ERK1/2 phosphorylation and IRS-1 phosphorylation at Ser616; these effects were reversed by ERK1/2 inhibition. We further observed that 20-HETE treatment resulted in impaired insulin-stimulated IRS-1 phosphorylation at Tyr632 and subsequent PI3-kinase/Akt activation. Furthermore, 20-HETE treatment blocked insulin-stimulated phosphorylation of eNOS at the stimulatory Ser1177 site, eNOS activation and NO production; these effects were reversed by inhibiting ERK1/2. Treatment of C57BL/6J mice with 20-HETE resulted in ERK1/2 activation and impaired insulin-dependent activation of the IRS-1/PI3K/Akt/eNOS pathway in the aorta. Our data suggest that the 20-HETE activation of IRS-1 phosphorylation at Ser616 is dependent on ERK1/2 and leads to impaired insulin-stimulated vasodilator effects that are mediated by the IRS-1/PI3K/AKT/eNOS pathway.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Aorta / drug effects
Aorta / metabolism
Cardiovascular Agents / pharmacology*
Cardiovascular Diseases / drug therapy
Cardiovascular Diseases / metabolism
Cells, Cultured
Drug Evaluation, Preclinical
Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
Extracellular Signal-Regulated MAP Kinases / metabolism
Human Umbilical Vein Endothelial Cells / drug effects
Human Umbilical Vein Endothelial Cells / metabolism*
Humans
Hydroxyeicosatetraenoic Acids / pharmacology*
Insulin / physiology*
Insulin Receptor Substrate Proteins / metabolism*
MAP Kinase Signaling System
Mice, Inbred C57BL
Nitric Oxide / metabolism
Nitric Oxide Synthase Type III / metabolism
Phosphatidylinositol 3-Kinases / metabolism
Phosphorylation
Protein Processing, Post-Translational
Proto-Oncogene Proteins c-akt / metabolism
Serine / metabolism

Substances

Cardiovascular Agents
Hydroxyeicosatetraenoic Acids
IRS1 protein, human
Insulin
Insulin Receptor Substrate Proteins
Nitric Oxide
Serine
20-hydroxy-5,8,11,14-eicosatetraenoic acid
NOS3 protein, human
Nitric Oxide Synthase Type III
Phosphatidylinositol 3-Kinases
Proto-Oncogene Proteins c-akt
Extracellular Signal-Regulated MAP Kinases

Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 81300090 and No. 30930039,http://www.nsfc.gov.cn/) and the Young Physician Training Project of Shanghai. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.