Influence of endothelial nitric oxide synthase haplotypes on nitric oxide and peroxynitrite productions

Seham O Alsulami; Tadeusz Malinski; Howard D Dewald

doi:10.1016/j.bioelechem.2024.108837

Influence of endothelial nitric oxide synthase haplotypes on nitric oxide and peroxynitrite productions

Bioelectrochemistry. 2025 Feb:161:108837. doi: 10.1016/j.bioelechem.2024.108837. Epub 2024 Oct 16.

Authors

Seham O Alsulami¹, Tadeusz Malinski², Howard D Dewald³

Affiliations

¹ Department of Chemistry and Biochemistry, Ohio University, Athens, OH, USA; Department of Biochemistry, Faculty of Sciences, University of Tabuk, Tabuk, Saudi Arabia.
² Department of Chemistry and Biochemistry, Ohio University, Athens, OH, USA; Deceased September 13, 2023.
³ Department of Chemistry and Biochemistry, Ohio University, Athens, OH, USA. Electronic address: dewald@ohio.edu.

PMID: 39426051
DOI: 10.1016/j.bioelechem.2024.108837

Abstract

The impact of four clinically significant genetic variants of endothelial nitric oxide synthase (eNOS) polymorphisms on the concentrations of nitric oxide [NO] and peroxynitrite [ONOO^-] has been given scant consideration. This study utilized a [NO]/[ONOO^-] ratio to determine the extent of endothelial dysfunction caused by these variations in the eNOS gene. The single nucleotide polymorphisms (T-786C, C-665T, and Glu298Asp) and a variable number of tandem repeats (intron 4 a/b/c) were genotyped in human umbilical vein endothelial cells (HUVEC), using sanger sequencing and DNA electrophoresis, respectively. Nanosensors were used to determine the maximal [NO] and [ONOO^-], while traditional and low-temperature SDS-PAGE were used to evaluate the expression of eNOS and the eNOS dimer-to-monomer ratio, respectively. The study results indicate that the eNOS haplotype H3 (G T/C C 4a/c allele) may have a protective effect against cardiovascular disease (CVD) with the [NO]/[ONOO^-] ratio higher than 2. However, the eNOS haplotypes H2 (G T/C C 4a/b) and H5 (T T/C C 4b) increase the susceptibility to CVD with [NO]/[ONOO^-] ratio lower than 1. The results suggest that certain eNOS genetic variants may influence susceptibility to cardiovascular disease (CVD) while other variants may have a protective effect.

Keywords: Endothelial dysfunction; Nanosensor; eNOS gene; eNOS monomer/dimer; eNOS uncoupling.

MeSH terms

Haplotypes*
Human Umbilical Vein Endothelial Cells* / metabolism
Humans
Nitric Oxide Synthase Type III* / genetics
Nitric Oxide Synthase Type III* / metabolism
Nitric Oxide* / metabolism
Peroxynitrous Acid* / metabolism
Polymorphism, Single Nucleotide*

Substances

Nitric Oxide Synthase Type III
Peroxynitrous Acid
Nitric Oxide
NOS3 protein, human