Tacconelli Stefania, Capone Marta L, Patrignani Paola
Department of Medicine and Center of Excellence on Aging, G. d'Annunzio University School of Medicine, Chieti, Italy.
Methods Mol Biol. 2010;644:165-78. doi: 10.1007/978-1-59745-364-6_14.
Several lines of evidence suggest that reactive oxygen species are implicated in human disease, including atherosclerosis, hypertension, and restenosis after angioplasty. The measurement of F(2)-isoprostanes (F(2)-iPs), formed nonenzymatically through free radical catalyzed attack on esterified arachidonate, provides a reliable tool for identifying populations with enhanced rates of lipid peroxidation. Among F(2)-isoPs, 8-iso-PGF(2alpha) (also referred to IPF(2alpha)-III) and IPF(2alpha)-VI are the most frequently measured in biological fluids. A variety of methods have been proposed to measure F(2)-isoprostanes in urine and plasma. Mass spectrometry has been developed for the measurement of both F(2)-isoprostanes but its use is limited as it is time-consuming and highly expensive. We have developed validated enzyme immunoassay (EIA) and radioimmunoassay (RIA) techniques using highly specific antisera for the measurement of 8-iso-PGF(2alpha). In contrast, the commercially available immunoassay kits are limited for their poor specificity. The measurement of specific isoprostanes, such as 8-iso-PGF(2alpha), in urine is a reliable, noninvasive index of lipid peroxidation that is of valuable help in dose-finding studies of natural and synthetic antioxidant agents.
多项证据表明，活性氧与人类疾病有关，包括动脉粥样硬化、高血压以及血管成形术后再狭窄。通过自由基催化攻击酯化花生四烯酸非酶促形成的F(2)-异前列腺素（F(2)-iPs）的测量，为识别脂质过氧化速率增加的人群提供了一种可靠工具。在F(2)-异前列腺素中，8-异前列腺素F(2α)（也称为IPF(2α)-III）和IPF(2α)-VI是生物流体中最常测量的。已经提出了多种方法来测量尿液和血浆中的F(2)-异前列腺素。质谱法已被开发用于测量这两种F(2)-异前列腺素，但由于其耗时且成本高昂，其应用受到限制。我们已经开发了经过验证的酶免疫测定（EIA）和放射免疫测定（RIA）技术，使用高度特异性抗血清来测量8-异前列腺素F(2α)。相比之下，市售的免疫测定试剂盒因其特异性差而受到限制。尿液中特定异前列腺素（如8-异前列腺素F(2α)）的测量是脂质过氧化的可靠、非侵入性指标，在天然和合成抗氧化剂的剂量研究中具有重要帮助。
