Radioimmunological measurement of F(2)-isoprostanes after hydrolysis of lipids in tissues.
8-Iso-prostaglandin F(2 alpha)(8-iso-PGF(2 alpha)) is a major isoprostane formed in vivo mainly by non-enzymatic peroxidation of arachidonic acid and a potential biomarker of oxidative injury. We have recently reported development of a specific radioimmunoassay for the measurement of free 8-iso-PGF(2 alpha)in plasma and urine. The aim of this study was to employ this radioimmunoassay to analyze the total amount of 8-iso-PGF(2 alpha)(sum of free and esterified) in liver tissues by using alkaline hydrolysis, and to apply it in an experimental model of carbon tetrachloride-induced lipid peroxidation in rats. Basal levels of total 8-iso-PGF(2 alpha)in hydrolyzed liver tissues of control rats were 6.5 times higher than the levels of free 8-iso-PGF(2 alpha). At maximum formation of total 8-iso-PGF(2 alpha)in the livers of carbon tetrachloride-treated rats, total levels of 8-iso-PGF(2 alpha)were almost 13 times higher than the levels of free 8-iso-PGF(2 alpha). In conclusion, high levels of 8-iso-PGF(2 alpha)in tissues can be quantified after hydrolysis both at basal conditions and in a model of increased lipid peroxidation. The methodology for measurement of total levels of 8-iso-PGF(2 alpha)in tissues may be suitable for future investigations of the location of oxidative injury in the body.
Södergren
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E
Department of Public Health and Caring Sciences, Section for Geriatrics/Clinical Nutrition Research, Uppsala University, Sweden.
Vessby
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B
Basu
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S
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D015415:Biomarkers; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism*; D028441:F2-Isoprostanes; D006868:Hydrolysis; D050356:Lipid Metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D018384:Oxidative Stress; D011863:Radioimmunoassay / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D015415:Biomarkers; D028441:F2-Isoprostanes; C075750:8-epi-prostaglandin F2alpha; D016718:Arachidonic Acid; D015237:Dinoprost