Related Experiment Video
Updated: Dec 21, 2025

Preparation of Rat Skeletal Muscle Homogenates for Nitrate and Nitrite Measurements
Published on: July 29, 2021
Nitric Oxide Modulation by Folic Acid Fortification
1Department Bioresources Engendering, Okinawa College, National Institute of Technology, 905 Henoko, Nago, Okinawa 905-2192, Japan.
Abstract:
Folic acid (FA) can be protected the neural tube defects (NTDs) causing nitric oxide (NO) induction, but the alleviation mechanism of the detailed FA function against NO has not yet been clarified. This study focused on elucidation of the interaction of FA and NO. FA suppressed nitrite accumulation as the NO indicator in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, then the expression of the iNOS gene due to the LPS treatment was not inhibited by FA, suggesting that FA can modulate against NO or nitrogen radicals. NOR3 (4-Ethyl-2-hydroxyamino-5-nitro-3-hexenamide) as the NO donor was used for evaluation of the NO scavenging activity of FA. FA suppressed the nitrite accumulation in a dose-dependent manner. To confirm the reaction product of FA and NO (FA-NO), liquid chromatography-mass spectrometry (LC/MS) was used to measure a similar system containing NOR3 and FA, and then detected the mass numbers of the FA-NO as m/z 470.9 (M + H)+ and m/z 469.1 (M - H)-. In addition, the adducts of the FA-NO derived from 14NO and 15NO gave individual mass numbers of the isotopic ratio of nitrogen for the following products: FA-14NO, m/z 471.14 (M + H)+; m/z 469.17 (M - H)- and FA-15NO, m/z 472.16 (M + H)+; m/z 470.12 (M - H)-. To clarify the detailed NO scavenging action of FA, an electron spin resonance (ESR) study for radical detecting of the system containing carboxy-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) as an NO detection reagent in the presence of NOR3 and FA was performed. The carboxy-PTI (2-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl) radical produced from the reaction with NO reduced in the presence of FA showing that FA can directly scavenge NO. These results indicated that NO scavenging activity of FA reduced the accumulation of nitrite in the LPS-stimulated RAW264.7 cells. The NO modulation due to FA would be responsible for the alleviation from the failure in neural tube formation causing a high level of NO production.
More Related Videos
Related Concept Videos
Nitric Oxide Signaling Pathway
2° Amines to N-Nitrosamines: Reaction with NaNO2
Overview of Nitrogen Metabolism
The largest pool of nitrogen available in the terrestrial ecosystem is gaseous nitrogen (N2) from the air, but this...
Key Elements for Plant Nutrition
Inorganic Nitrogen Assimilation
Regulation of Angiogenesis and Blood Supply

