Measurement of isoprostanes as markers of oxidative stress in neuronal tissue

Abstract

Oxidative stress is implicated in the pathogenesis of a variety of human diseases, including neurodegenerative disease, atherosclerosis and cancer, as well as progressive and even normal aging processes. Increased generation of free radicals derived primarily from molecular oxygen has also been associated with neuronal damage induced by a variety of environmental agents. However, measuring oxidative stress in biological systems is complex and requires accurate quantification of either free radicals or damaged biomolecules. One method to quantify oxidative injury is to measure lipid peroxidation. Lipids are readily attacked by free radicals, resulting in the formation of a number of peroxidation products. F₂-isoprostanes (F₂-IsoPs) are one group of these compounds, which are derived by the free radical peroxidation of arachidonic acid (AA). The F₂-IsoPs, prostaglandine F₂-like compounds, have been shown as the most accurate measure of oxidative damage in vivo. This review summarizes current methodology used to quantify F₂-IsoPs and discusses the utility of these and other prostaglandine (PG)-like compounds as in vivo biomarkers of oxidative stress in neuronal tissues.

Keywords: F2-isoprostanes; lipid peroxidation; neuroprostanes; oxidative damage.

Figure 1

Schematic presentation of procedure used for the extraction, purification, derivatization and mass spectrometric analysis of F₂-IsoPs from neuronal tissues

Figure 2

Chromatograms for F₂-IsoPs and F₄-NeuroPs from the mouse cerebrum. All chromatograms plot abundance vs. time (min). (A) m/z 569 chromatogram showing F₂-IsoPs, (B) m/z 573 chromatogram showing internal standard, (C) m/z 593 chromatogram showing F₄-NeuroPs.

Figure 3

Time course for cerebral oxidative damage following ICV injection of LPS. Data are F₂-IsoPs and F₄-NeuroPs expressed as ng per g tissue following injection at time = 0 (n≥5 per time point). * p<0.01 compared to untreated control.