Long lasting protective effects of early l-arginine treatment on endothelium in an in vitro study

Clin Nutr. 2021 Apr;40(4):1519-1529. doi: 10.1016/j.clnu.2021.02.040. Epub 2021 Mar 3.


Background & aims: Excess nutrient supply, such as high fat and high glucose intake, promotes oxidative stress and advanced glycation end products accumulation. Oxidative stress and AGE accumulation cause pathological elevation of arginase activity and pro-inflammatory signaling implicated in endothelial dysfunction. Several studies showed positive effects of l-arginine supplementation in endothelial function but little is currently known about the role of l-arginine as prevention of endothelial dysfunction caused by excessive nutrient supply (overfeeding). Our aim was to evaluate a possible protective effect of l-arginine on endothelial dysfunction caused by excessive nutrient supply (overfeeding), using human endothelial cells line in an in vitro study.

Methods: Endothelial EA.hy926 cells were pre-treated with 1.72 mM of l-arginine for 24 h and afterwards subjected to nutritional stress (high lipid, high insulin and high glucose concentrations) for further 24 h. After treatment discontinuation, the cells were kept in culture for 48 h, in physiological condition, to evaluate the effects of treatments after normalization.

Results: Excess nutrient supply in EA.hy926 cell line showed an increase of oxidative and nitrosative stress, a rise of AGEs production, high arginase activity, leading the cells to acidosis and to cell death. l-arginine pretreatment protects the cells by reducing apoptosis, acidosis, oxidative and nitrosative stress, arginase activity and AGE accumulation. l-arginine pretreatment reduces AGEs generation and accumulation by regulating STAB1 and RAGE gene expression levels. STAB1, acting as receptor scavenger of AGEs, interferes with AGE-RAGE binding and thus prevents activation of intracellular signaling pathways leading to cell damage. Moreover the reduction of oxidative stress promotes a decrease of excessive activation of arginase involved in endothelial dysfunction. The effects of pretreatment with l-arginine last even in the absence of stimuli and despite after treatment discontinuation.

Conclusions: An early l-arginine treatment is able to prevent oxidative stress and AGEs accumulation caused by overfeeding in human endothelial cell line by regulating STAB1/RAGE gene expression and by reducing excess arginase activity. The positive effects of l-arginine pretreatment continue even after treatment discontinuation in normal conditions.

Keywords: Endothelial EA.hy926 cells; Endothelial dysfunction; Excessive nutrient supply; Oxidative stress; Stabilin 1; l-arginine.

MeSH terms

  • Arginine / pharmacology*
  • Cell Line
  • Endothelial Cells / drug effects
  • Endothelium, Vascular / drug effects*
  • Glycation End Products, Advanced / metabolism
  • Humans
  • Nutritional Physiological Phenomena / drug effects*
  • Overnutrition / metabolism
  • Overnutrition / prevention & control*
  • Oxidative Stress / drug effects
  • Protective Agents / pharmacology*
  • Signal Transduction / drug effects


  • Glycation End Products, Advanced
  • Protective Agents
  • Arginine