Thu Vu Thi, Kim Hyoung Kyu, Long Le Thanh, Nyamaa Bayalagmaa, Song In-Sung, Thuy To Thanh, Huy Nguyen Quang, Marquez Jubert, Kim Soon Ha, Kim Nari, Ko Kyung Soo, Rhee Byoung Doo, Han Jin
National Research Laboratory for Mitochondrial Signaling, Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 47392, Korea.; Faculty of Biology, VNU University of Science, Hanoi 120036, Vietnam.
National Research Laboratory for Mitochondrial Signaling, Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 47392, Korea.
Korean J Physiol Pharmacol. 2016 Mar;20(2):201-11. doi: 10.4196/kjpp.2016.20.2.201. Epub 2016 Feb 23.
Although the antioxidant and cardioprotective effects of NecroX-5 on various in vitro and in vivo models have been demonstrated, the action of this compound on the mitochondrial oxidative phosphorylation system remains unclear. Here we verify the role of NecroX-5 in protecting mitochondrial oxidative phosphorylation capacity during hypoxia-reoxygenation (HR). Necrox-5 treatment (10 µM) and non-treatment were employed on isolated rat hearts during hypoxia/reoxygenation treatment using an ex vivo Langendorff system. Proteomic analysis was performed using liquid chromatography-mass spectrometry (LC-MS) and non-labeling peptide count protein quantification. Real-time PCR, western blot, citrate synthases and mitochondrial complex activity assays were then performed to assess heart function. Treatment with NecroX-5 during hypoxia significantly preserved electron transport chain proteins involved in oxidative phosphorylation and metabolic functions. NecroX-5 also improved mitochondrial complex I, II, and V function. Additionally, markedly higher peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC1α) expression levels were observed in NecroX-5-treated rat hearts. These novel results provide convincing evidence for the role of NecroX-5 in protecting mitochondrial oxidative phosphorylation capacity and in preserving PGC1α during cardiac HR injuries.
尽管NecroX-5在各种体外和体内模型中的抗氧化和心脏保护作用已得到证实，但其对线粒体氧化磷酸化系统的作用仍不清楚。在此，我们验证了NecroX-5在缺氧复氧（HR）过程中保护线粒体氧化磷酸化能力的作用。在使用离体Langendorff系统进行缺氧/复氧处理期间，对分离的大鼠心脏采用Necrox-5处理（10 µM）和不处理。使用液相色谱-质谱（LC-MS）和非标记肽计数蛋白质定量进行蛋白质组学分析。然后进行实时PCR、蛋白质印迹、柠檬酸合酶和线粒体复合物活性测定以评估心脏功能。缺氧期间用NecroX-5处理可显著保留参与氧化磷酸化和代谢功能的电子传递链蛋白。NecroX-5还改善了线粒体复合物I、II和V的功能。此外，在NecroX-5处理的大鼠心脏中观察到明显更高的过氧化物酶体增殖物激活受体γ共激活因子-1α（PGC1α）表达水平。这些新结果为NecroX-5在心脏HR损伤期间保护线粒体氧化磷酸化能力和保留PGC1α的作用提供了令人信服的证据。
