Physical Chemistry for Life Science Laboratory, Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan.
Department of Pharmacology, Faculty of Science, Mahidol University, Rama 6 Road, Ratchathewi, Bangkok 10400, Thailand.
Free Radic Biol Med. 2017 Dec;113:487-493. doi: 10.1016/j.freeradbiomed.2017.10.388. Epub 2017 Oct 28.
Oxidized low density lipoprotein (Ox-LDL) is implicated in a variety of oxidative diseases. To clarify the mechanisms involved and facilitate the investigation of therapeutics, we previously developed a detection method for lipid-derived radicals using the fluorescent probe 2,2,6-trimethyl-6-pentyl-4-(4-nitrobenzo[1,2,5]oxadiazol-7-ylamino)piperidine-1-oxyl (NBD-Pen). In this study, NBD-Pen was used to detect lipid-derived radicals in Ox-LDL from in vitro and in vivo samples using an iron overloaded mouse model. By following the timeline of lipid radical generation using this method, the iron overloaded mice could be successfully treated with the antioxidant Trolox, resulting in successful lowering of the plasma lipid peroxidation, aspartate transaminase and alanine transaminase levels. Furthermore, using a combination therapy of the chelating agent deferoxamine (DFX) and Trolox, liver injury and oxidative stress markers were also reduced in iron overloaded mice. The NBD-Pen method is highly sensitive as well as selective and is suitable for targeting minimally modified LDL compared with other existing methods.
氧化型低密度脂蛋白（Ox-LDL）与多种氧化性疾病有关。为了阐明相关机制并促进治疗方法的研究，我们先前开发了一种使用荧光探针 2,2,6-三甲基-6-戊基-4-(4-硝基苯并[1,2,5]恶二唑-7-基氨基)哌啶-1-氧自由基（NBD-Pen）检测脂质衍生自由基的方法。在这项研究中，使用铁超载小鼠模型，使用 NBD-Pen 从体外和体内样本中检测 Ox-LDL 中的脂质衍生自由基。通过使用这种方法跟踪脂质自由基生成的时间线，抗氧化剂 Trolox 可以成功治疗铁超载小鼠，从而成功降低血浆脂质过氧化、天冬氨酸转氨酶和丙氨酸转氨酶水平。此外，使用螯合剂去铁胺（DFX）和 Trolox 的联合治疗，铁超载小鼠的肝损伤和氧化应激标志物也减少。与其他现有方法相比，NBD-Pen 方法具有高灵敏度和选择性，并且适合于靶向最小修饰的 LDL。
