Radioimmunosassay of 8-iso-prostaglandin F2α: an index for oxidative injury via free radical catalysed lipid peroxidation

doi:10.1016/S0952-3278(98)90042-4

Prostaglandins, Leukotrienes and Essential Fatty Acids

Volume 58, Issue 4, April 1998, Pages 319-325

https://doi.org/10.1016/S0952-3278(98)90042-4 Get rights and content

Abstract

8-Iso-prostaglandin F_2α (8-iso-PGF_2α), a major F₂-isoprostane, is biosynthesized in vivo through nonenzymatic free radical-catalysed peroxidation of arachidonic acid. The levels of F₂-isoprostanes both free in the circulation and esterified to the tissue phospholipids increase intensely in animal models of oxidant injury. This study presents the development and validation of a radioimmunoassay of 8-iso-PGF_2α for the measurement of this substance in the body fluids. Furthermore, its application in normal human volunteers, a pharmacokinetic study performed in rabbits with 8-iso-PGF_2α, and hepatic lipid peroxidation in rats is reported. An antibody was raised in rabbits by immunization with 8-iso-PGF_2α coupled to BSA at the carboxylic acid by 1,1′-Carbonyldiimmidazole method. The cross-reactivity of the antibody with 8-iso-15-keto-13,14-dihydro-PGF_2α, 8-iso-PGF_2β, PGF_2α, 15-keto-PGF_2α, 15-keto-13,14-dihydro-PGF_2α,TXB₂, 11β-PGF_2α, 9β-PGF_2α and 8-iso-PGF_3α was 1.7, 9.8, 1.1, 0.01, 0.01, 0.1, 0.03, 1.8 and 0.6%, respectively. The intraassay precision was 14.5% (CV) at the level of 64 pg/0.1 ml and 12.2% with 512 pg/0.1 ml in the human plasma. Similarly, intra-assay accuracy was 95.6% and 101% for the low and the high standard, respectively. The detection limit was about 23 pmol/l. The appearance and disappearance of 8-iso-PGF_2α in the blood and urine following intravenous administration of 8-iso-PGF_2α in the rabbit was rapid. Free 8-iso-PGF_2α levels in plasma and urine from normal human volunteers are evaluated and found to correlate with the obtained values by gas chromatography-mass spectrometry methods from other studies. The levels of free 8-iso-PGF_2α in the plasma and urine increased 7- and 102-fold, respectively, after CCI₄ administration to rats. Thus, this 8-iso PGF_2α radioimmunoassay method is relevant to apply in the oxidative injury studies as an index of in vivo lipid peroxidation through free radical catalysis mechanism.

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Citation Excerpt :
F2-IsoPs are relatively stable in urine (Roberts and Morrow, 2000) and can be assessed by two main analytical approaches: mass-based techniques (using either gas – GC or liquid – LC chromatography) and immunological techniques. GC–MS is the gold standard for the analysis of F2-IsoPs; however, immunological techniques are widely used to measure 8-isoprostane, because radioimmunoassay and ELISA kits have been developed for rapid detection and quantification of 8-isoprostane (Basu, 1998; Ito et al., 2019). Studies, which compared both methods, found that 8-isoprostane concentrations determined with chromatography/mass spectrometry-based assays did not agree with those determined with immunoassays, suggesting that GC–MS and EIA do not measure the same compounds.
Oxidative stress is defined as an imbalance between the production and elimination of reactive oxygen species (ROS) are associated with various inflammation-related human disease. ROS can oxidize lipids, which subsequently undergo fragmentation to produce F2-isoprostanes (F2-IsoPs). Eight-isoprostane is one of the most extensively studied F2-IsoPs and the most commonly used biomarker for the assessment of oxidative stress in human studies. This urinary biomarker is quantified using either chemical or immunological techniques. A “physiological” range for 8-isoprostanes is needed to use this biomarker as a measure of excess oxidative stress originating from occupational exposures. However, ranges reported in the literature are inconsistent. We designed a standardized protocol of a systematic review and meta-analysis to assess baseline values for 8-isoprostane concentrations in urine of healthy adults and identify determinants of their inter- and intra-individual variability. We searched PubMed from journal inception and up to April 2019, and screened articles for studies containing F2-IsoPs concentrations in urine for healthy adult participants. We grouped studies in three biomarker groups: “8-isoprostane”, “Isoprostanes” “15- F2t-Isoprostane”. We computed geometric mean (GM) and geometric standard deviation (GSD) as the basis for the meta-analysis. Of the initial 1849 articles retrieved, 63 studies were included and 107 subgroups within these study populations were identified. We stratified the subgroups analyzed with the chemical methods by body mass index (BMI) reported. We provide pooled GM values for urinary 8-isoprostane concentrations in healthy adults, separately for chemical and immunological analysis in this review. The interquartile range (IQR) in subgroups with a mean BMI below 25 measured using chemical methods was 0.18 to 0.40 μg/g creatinine. We show that there is a significant positive association between BMI and urinary 8-isoprostane concentrations. We recommend adjusting urinary 8-isoprostane concentrations in spot urine with creatinine, quantifying 8-isoprostane with chemical analytical methods, and reporting results as median and quartiles. This will help in comparing results across studies.
Biomarkers of Oxidative/Nitrosative Stress and Neurotoxicity
2019, Biomarkers in Toxicology
Oxidative stress is implicated as one of the major underlying mechanisms in a variety of human diseases. Excess formation of free radicals may overwhelm the capacity of endogenous cellular antioxidant defense mechanisms, cause alterations in normal cell and organ physiology, and activate and/or accelerate disease processes. Reactive radicals derived from molecular oxygen (reactive oxygen species, ROS) and nitrogen (reactive nitrogen species, RNS) readily attack a variety of critical biological molecules, including lipids, DNA, and essential cellular proteins. High content of unsaturated lipids in the brain leads to pronounced lipid peroxidation, a central feature of oxidant injury in the brain. Oxidative stress is also associated with cell response to a variety of toxicants. This chapter analyzes oxidative stress and the cell response to diverse toxicants and presents evidence of ROS/RNS generation and neurodegeneration in different toxicity models.
Development of an LC-ESI(-)-MS/MS method for the simultaneous quantification of 35 isoprostanes and isofurans derived from the major n3- and n6-PUFAs
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Misregulation of oxidative and antioxidative processes in the organism – oxidative stress – contributes to the pathogenesis of different diseases, e.g. inflammatory or neurodegenerative diseases. Oxidative stress leads to autoxidation of polyunsaturated fatty acids giving rise to prostaglandin-like isoprostanes (IsoP) and isofurans (IsoF). On the one hand they could serve as biomarker of oxidative stress and on the other hand may act as lipid mediators, similarly as the enzymatically formed oxylipins. In the present paper we describe the development of an LC-ESI(-)-MS/MS method allowing the parallel quantification of 27 IsoP and 8 IsoF derived from 6 different PUFA (ALA, ARA, EPA, AdA, n6-DPA, DHA) within 12 min. The chromatographic separation was carried out on an RP-C18 column (2.1 × 150 mm, 1.8 μm) yielding narrow peaks with an average width at half maximum of 3.3–4.2 s. Detection was carried out on a triple quadrupole mass spectrometer operating in selected reaction monitoring mode allowing the selective detection of regioisomers. The limit of detection ranged between 0.1 and 1 nM allowing in combination with solid phase extraction the detection of IsoP and IsoF at subnanomolar concentrations in biological samples. The method was validated for human plasma showing high accuracy and precision. Application of the approach on the investigation of oxidative stress in cultured cells indicated a distinct pattern of IsoP and IsoF in response to reactive oxygen species which warrants further investigation.
The described method is not only the most comprehensive approach for the simultaneous quantification of IsoP and IsoF, but it was also integrated in a targeted metabolomics method (Ostermann et al. (2015) Anal Bioanal Chem) allowing the quantification of in total 164 oxylipins formed enzymatically and non-enzymatically within 30.5 min.
Inflammatory F<inf>2</inf>-isoprostane, prostaglandin F<inf>2α</inf>, pentraxin 3 levels and breast cancer risk: The Swedish Mammography Cohort
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Citation Excerpt :
Controls were randomly selected from women without a breast cancer diagnosis who had provided a blood and urine sample. Urinary samples were analysed for 8-iso-PGF2α as an index of oxidative injury by a validated radioimmunoassay [29]. Urinary samples were also analysed for 15-keto-dihydro PGF2α, major metabolite of PGF2α, as an index of inflammatory response by a validated radioimmunoassay [30].
Breast cancer is a common cancer among women. Identifying cellular participation of F₂-isoprostane, prostaglandin F_2α (PGF_2α) and pentraxin 3 (PTX3) in cancer we evaluated whether their prediagnostic systemic levels that originate from different inflammatory pathways were associated with breast cancer risk.
Seventy-eight breast cancer cases diagnosed after blood collection and 797 controls from the Swedish Mammography Cohort were analysed for urinary F₂-isoprostane, PGF_2α and plasma PTX3 levels.
None of the biomarkers investigated were significantly associated with breast cancer risk. However, there was the suggestion of an inverse association with PTX3 with multivariable adjusted ORs (95% CI) of 0.56 (95% CI=0.29–1.06) and 0.67 (95% CI=0.35–1.28) for the second and third tertiles, respectively (p_trend=0.20). No associations were observed between F₂-isoprostane (OR=0.87; 95% CI=0.48–1.57; p_trend=0.67) and PGF_2α metabolite (OR=1.03; 95% CI=0.56–1.88; p_trend=0.91) comparing the top to bottom tertiles.
The systemic levels of F₂-isoprostane, PGF_2α and PTX3 witnessed in women who later developed breast cancer may not provide prognostic information regarding tumor development in spite of their known involvement in situ cellular context. These observations may indicate that other mechanisms exist in controlling cellular formation of F₂-isoprostane, PGF_2α and PTX3 and their systemic availability in breast cancer patients.

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Radioimmunosassay of 8-iso-prostaglandin F2α: an index for oxidative injury via free radical catalysed lipid peroxidation

Abstract

Trends Biochem Sci

FEBS Lett

Biochem Pharmacol

Methods Enzymol

Prostaglandins Leukot Essent Fatty Acids

J Biol Chem

Free Radic Biol Med

Hepatology

Free radicals in biology and medicine

Free radical mechanisms in tissue injury

Biochem J

Role of oxidative stress in development of complications in diabetes

Diabetes

Protein oxidation and aging

Science

The measurement of free radical reactions in humans

Some thoughts for future experimentation

FEBS Lett

Protein oxidation and aging

Difficulties in measuring reactive protein carbonyls in tissue using 2-4-dinitrophenylhydrazine

Arch Biochem Biophys

Radioimmunosassay of 8-iso-prostaglandin F_2α: an index for oxidative injury via free radical catalysed lipid peroxidation