Wiswedel Ingrid
Department of Pathological Biochemistry, Medical Faculty of the Otto-von-Guericke-University, Magdeburg, Germany.
Methods Mol Biol. 2009;580:3-16. doi: 10.1007/978-1-60761-325-1_1.
A gas chromatography-mass spectrometric method was developed that allowed the accurate, highly sensitive and specific quantification of F(2)-isoprostanes (F(2)-IsoPs) in different tissues and body fluids. Measurement of F(2)-IsoPs in isolated rat brain mitochondria, HaCaT keratinocytes, human plasma, and microdialysates of human skin has established the occurrence of oxidative stress in a variety of model systems and disease states. F(2)-IsoPs correlated with other markers of lipid peroxidation (e.g., TBARS, HETEs) in experimental models of oxidative stress. F(2)-IsoPs were elevated about 100-fold after iron/ascorbate-induced oxidative stress and 2- to 4-fold after pentylenetetrazol (PTZ)-induced seizures, in hemodialysis patients with end stage renal disease, in psoriasis patients, in HaCaT keratinocytes, and in microdialysates of human skin following UVB irradiation.Both human and experimental studies have indicated associations of F(2)-IsoPs and inflammatory conditions. Anti-inflammatory drugs such as diclofenac did not only suppress the prostaglandin but also the F(2)-IsoP pathway.Microdialysis allows the "near-in vivo" measurement of prostanoid mediators, released in the interstitial space of the dermis under inflammatory conditions.
开发了一种气相色谱 - 质谱法,可对不同组织和体液中的F(2)-异前列腺素(F(2)-IsoPs)进行准确、高度灵敏且特异的定量分析。对分离的大鼠脑线粒体、HaCaT角质形成细胞、人血浆以及人皮肤微透析液中的F(2)-IsoPs进行测定,证实了在多种模型系统和疾病状态下均存在氧化应激。在氧化应激实验模型中,F(2)-IsoPs与脂质过氧化的其他标志物(如硫代巴比妥酸反应物、羟基二十碳四烯酸)相关。在铁/抗坏血酸诱导的氧化应激后,F(2)-IsoPs升高约100倍;在戊四氮(PTZ)诱导的癫痫发作后、终末期肾病血液透析患者、银屑病患者、HaCaT角质形成细胞以及紫外线B(UVB)照射后人皮肤微透析液中,F(2)-IsoPs升高2至4倍。人体研究和实验研究均表明F(2)-IsoPs与炎症状态有关。双氯芬酸等抗炎药物不仅抑制前列腺素途径,还抑制F(2)-IsoP途径。微透析能够“近体内”测量在炎症条件下真皮间质空间释放的类前列腺素介质。