Analysis of Oxidative Stress and Wound-Inducible Dinor Isoprostanes F1 (Phytoprostanes F1) in Plants

Abstract

Abstract Isoprostanes F2 are arachidonate autoxidation products in mammals that have been shown to be induced during several human disorders associated with enhanced free-radical generation. Isoprostanes F2 represent not only extremely reliable markers of oxidative stress in vivo, but they also exert potent biological effects. Therefore, it has been postulated that isoprostanoids are mediators of oxidant injury in vivo. Higher plants, however, do not synthesize arachidonic acid or isoprostanes. Here we show that a series of isoprostane F2 analogs termed phytoprostanes F1 (previously dinor isoprostanes F1) are formed by an analogous pathway from α-linolenate in plants. High-performance liquid chromatography and gas chromatography-mass spectrometry methods using [18O]3phytoprostanes F1 as internal standard have been developed to quantify phytoprostanes F1. In fresh peppermint (Mentha piperita) leaves, phytoprostanes F1 were found in free form (76 ng/g of dry weight) and at about 150-fold higher levels esterified in lipids. It is notable that these levels of phytoprostanes F1 are more than two orders of magnitude higher than the basal levels of isoprostanes F2 in mammalian tissues. Furthermore, wounding, as well as butyl hydroperoxide or cupric acetate stress triggered a dramatic increase of free and esterified phytoprostanes F1. Thus phytoprostanes F1 may represent a sensitive measure of oxidative damage in plants similar to isoprostanes in mammals. However, one of the most exciting issues to be clarified is the possibility that linolenate-derived phytoprostanes F1 exert biological activities in plants and/or animals.