Original Contributions
Measurement of F2-isoprostanes as an index of oxidative stress in vivo

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Abstract

In 1990 we discovered the formation of prostaglandin F2-like compounds, F2-isoprostanes (F2-IsoPs), in vivo by nonenzymatic free radical–induced peroxidation of arachidonic acid. F2-IsoPs are initially formed esterified to phospholipids and then released in free form. There are several favorable attributes that make measurement of F2-IsoPs attractive as a reliable indicator of oxidative stress in vivo: (i) F2-IsoPs are specific products of lipid peroxidation; (ii) they are stable compounds; (iii) levels are present in detectable quantities in all normal biological fluids and tissues, allowing the definition of a normal range; (iv) their formation increases dramatically in vivo in a number of animal models of oxidant injury; (v) their formation is modulated by antioxidant status; and (vi) their levels are not effected by lipid content of the diet. Measurement of F2-IsoPs in plasma can be utilized to assess total endogenous production of F2-IsoPs whereas measurement of levels esterified in phospholipids can be used to determine the extent of lipid peroxidation in target sites of interest. Recently, we developed an assay for a urinary metabolite of F2-IsoPs, which should provide a valuable noninvasive integrated approach to assess total endogenous production of F2-IsoPs in large clinical studies.

Section snippets

Biochemistry of the formation of isoprostanes

One of the greatest needs in the field of free radical research has been the availability of a reliable noninvasive approach to assess oxidative stress status in humans [1]. It has long been recognized that methods previously developed for this purpose lack specificity, sensitivity, or are too invasive for human investigation [2]. In 1990, we reported the discovery of the formation of prostaglandin F2-like compounds in vivo in humans by nonenzymatic free radical–induced peroxidation of

Methodology for measurement of F2-isoprostanes

Some discussion about the methodology used for measurement of F2-IsoPs is important. The initial discovery of F2-IsoPs was made possible by the use of mass spectrometric analysis. We, and others, continue to use stable isotope dilution gas chromatography negative ion chemical ionization mass spectrometry for measurement of F2-IsoPs [13]. Although mass spectrometric methodology is expensive and time consuming, it is highly specific and sensitive. The accuracy of our method of assay is 96% and

Favorable characteristics of F2-isoprostanes as a measure of oxidative stress in vivo

The discovery of the formation of F2-IsoPs was initially of biochemical interest but evolving from subsequent studies is the notion that measurement of these compounds likely represents one of the most reliable approaches to assess oxidative stress status in vivo. There are a number of favorable attributes that imply that measurement of F2-IsoPs may provide a reliable marker of lipid peroxidation in vivo (Table 1). First, these are stable compounds. F2-IsoPs are also specific products of free

Direct comparison of measuring F2-IsoPs with other measures of lipid peroxidation to assess oxidant injury in vitro and in vivo

To explore whether the value of measuring F2-IsoPs surpasses that of some routinely used measures of lipid peroxidation in vivo, we directly compared measurements of F2-IsoPs with malondialdehyde (MDA) and lipid hydroperoxides, commonly used measures of lipid peroxidation, both in vitro and in vivo. The results obtained were most revealing.

Various approaches to assess endogenous production of F2-IsoPs

There are several approaches that can be utilized to assess endogenous production of F2-IsoPs, each of which has certain advantages and/or drawbacks.

Summary and conclusions

In summary, initially the discovery of F2-IsoPs was primarily of biochemical interest. This discovery, however, has evolved over the last several years in a number of areas. These include the discovery of a number of additional classes of compounds that are generated as products of the IsoP pathway and the findings that several of these compounds can exert potent biological activity either through receptor-mediated actions or in the case of A2/J2-IsoPs and isolevuglandins because of their

Acknowledgements

Supported by grants GM42056, GM15431, DK48831, CA77839, DK26657, and CA68485 from the National Institutes of Health. J. D. M. is a recipient of a Burroughs Wellcome Fund Clinical Scientist Award in Translational Research.

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    1

    L. Jackson Roberts, II, received his M.D. degree from the University of Iowa and was elected to Alpha Omega Alpha. He did an Internal Medicine residency at Washington University, a fellowship in Clinical Pharmacology at Vanderbilt University, and was a recipient of a Burroughs Wellcome Scholar in Clinical Pharmacology Award. He is currently Professor of Pharmacology and Medicine at Vanderbilt University.

    2

    Jason D. Morrow received his M.D. degree from Washington University. He did an Internal Medicine residency at Vanderbilt University, an Infectious Disease fellowship at Washington University, and a Clinical Pharmacology fellowship at Vanderbilt University. He is currently a Professor of Medicine and Pharmacology at Vanderbilt University. He was a Howard Hughes Medical Institute Physician Research Fellow and the recipient of an NIH Physician Scientist Award, a International Life Sciences Institute Career Development Award, and a Burroughs Wellcome Fund Clinical Scientist Award in Translational Research.

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