The Role of Palmitic Acid in the Co-Toxicity of Bacterial Metabolites to Endothelial Cells

Abstract

Introduction

Metabolic endotoxemia most often results from obesity and is accompanied by an increase in the permeability of the intestinal epithelial barrier, allowing co-absorption of bacterial metabolites and diet-derived fatty acids into the bloodstream. A high-fat diet (HFD) leading to obesity is a significant extrinsic factor in developing vascular atherosclerosis. In this study, we evaluated the effects of palmitic acid (PA) as a representative of long-chain saturated fatty acids (LCSFA) commonly present in HFDs, along with endotoxin (LPS; lipopolysaccharide) and uremic toxin indoxyl sulfate (IS), on human vascular endothelial cells (HUVECs).

Methods

HUVECs viability was measured based on tetrazolium salt metabolism, and cell morphology was assessed with fluorescein-phalloidin staining of cells’ actin cytoskeleton. The effects of simultaneous treatment of endothelial cells with PA, LPS, and IS on nitro-oxidative stress in vascular cells were evaluated quantitatively with fluorescent probes. The expression of vascular cell adhesion molecule VCAM-1, E-selectin, and occludin, an essential tight junction protein, in HUVECs treated with these metabolites was evaluated in Western blot.

Results

PA, combined with LPS and IS, did not influence HUVECs viability but induced stress on actin fibers and focal adhesion complexes. Moreover, PA combined with LPS significantly enhanced reactive oxygen species (ROS) production in HUVECs but decreased nitric oxide (NO) generation. PA also considerably increased the expression of VCAM-1 and E-selectin in HUVECs treated with LPS or IS but decreased occludin expression.

Conclusion

Palmitic acid enhances the toxic effect of metabolic endotoxemia on the vascular endothelium.

Keywords:

• metabolic endotoxemia
• high-fat diet
• palmitic acid
• endotoxin
• indoxyl sulfate

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

The study was prepared under the project financed from the funds granted by the Ministry of Education and Science in Poland, in the “Regional Initiative of Excellence” program for the years 2019–2022, project number 016/RID/2018/19 (individual project number RID Z501.20.009).