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Related Experiment Video

Updated: May 9, 2026

Monitoring Equilibrium Changes in RNA Structure by 'Peroxidative' and 'Oxidative' Hydroxyl Radical Footprinting

Monitoring Equilibrium Changes in RNA Structure by 'Peroxidative' and 'Oxidative' Hydroxyl Radical Footprinting

Published on: October 17, 2011

Relationship between tissue hydroxyl radical and oxidatively modified macromolecule levels.

Toru Sasaki¹, Sho-Ichi Mogi, Takao Kaneko

¹Research Team for Mechanism of Aging, Redox Research, Tokyo Metropolitan Institute of Gerontology, Itabashi, Tokyo, Japan.

Geriatrics & Gerontology International

|July 25, 2013

Summary

This study optimized an in vivo method to measure hydroxyl radical (·OH) generation using salicylic acid (SA) hydroxylation. Hydroxyl radical levels increased with age, correlating with oxidative damage markers like 8-oxo-2'-deoxyguanosine (8-oxodG).

Keywords:

aging hydroxyl radical oxidatively modified macromolecules salicylic acid

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Area of Science:

Biochemistry
Aging Research
Oxidative Stress Biology

Background:

Hydroxyl radical (·OH) is a key reactive oxygen species implicated in cellular damage.
Oxidative modification of macromolecules is a hallmark of aging.
Quantifying ·OH generation in vivo remains a challenge.

Purpose of the Study:

To establish and optimize an in vivo method for measuring hydroxyl radical (·OH) generation.
To investigate the relationship between ·OH levels and macromolecule oxidation in young versus aged mice.
To assess age-related changes in oxidative stress markers.

Main Methods:

Optimized an in vivo ·OH detection method using salicylic acid (SA) hydroxylation.
Quantified 2,3-dihydroxybenzoic acid (2,3-DHBA) levels as a marker for ·OH.
Measured levels of 8-oxo-2 -deoxyguanosine (8-oxodG), carbonylated proteins, and thiobarbituric acid reactive substances (TBARS) in mouse tissues.

Main Results:

The optimized method demonstrated that 2,3-DHBA levels correlated with ·OH generation in tissues.
In young mice, 2,3-DHBA levels showed consistency with other oxidative stress markers (8-oxodG, carbonylated proteins).
Aging led to a biphasic increase in brain and heart 2,3-DHBA and 8-oxodG levels, indicating heightened oxidative stress.

Conclusions:

An optimized in vivo method using SA hydroxylation for ·OH measurement was developed.
The study confirmed correlations between 2,3-DHBA, 8-oxodG, and carbonylated proteins in young mice.
Aging significantly increases ·OH generation and oxidative damage, as evidenced by rising 2,3-DHBA and 8-oxodG levels.