作者: Wang, Zhi ; Melov, Simon ; Wong, Hoi-Shan ; Esbenshade, Tim ; Pliushchev, Marina ; Sweis, Ramzi F ; Brand, Martin D ; Riedmaier, Stephan ; Dighe, Pratiksha ; McKibben, Bryan ; Mezera, Vojtech ; Gerencser, Akos A

Superoxide produced by mitochondria has been implicated in numerous physiologies and pathologies. Eleven different mitochondrial sites that can produce superoxide and/or hydrogen peroxide (O₂^.-/H₂O₂) have been identified in vitro, but little is known about their contributions in vivo. We introduce novel variants of S1QELs and S3QELs (small molecules that suppress O₂^.-/H₂O₂ production specifically from mitochondrial sites I_Q and III_Qo, respectively, without compromising bioenergetics), that are suitable for use in vivo. When administered by intraperitoneal injection, they achieve total tissue concentrations exceeding those that are effective in vitro. We use them to study the engagement of sites I_Q and III_Qo in mice lacking functional manganese-superoxide dismutase (SOD2). Lack of SOD2 is expected to elevate superoxide levels in the mitochondrial matrix, and leads to severe pathologies and death about 8 days after birth. Compared to littermate wild-type mice, 6-day-old Sod2^-/- mice had significantly lower body weight, lower heart succinate dehydrogenase activity, and greater hepatic lipid accumulation. These pathologies were ameliorated by treatment with a SOD/catalase mimetic, EUK189, confirming previous observations. A 3-day treatment with S1QEL352 decreased the inactivation of cardiac succinate dehydrogenase and hepatic steatosis in Sod2^-/- mice. S1QEL712, which has a distinct chemical structure, also decreased hepatic steatosis, confirming that O₂^.- derived specifically from mitochondrial site I_Q is a significant driver of hepatic steatosis in Sod2^-/- mice. These findings also demonstrate the ability of these new S1QELs to suppress O₂^.- production in the mitochondrial matrix in vivo. In contrast, suppressing site III_Qo using S3QEL941 did not protect, suggesting that site III_Qo does not contribute significantly to mitochondrial O₂^.- production in the hearts or livers of Sod2^-/- mice. We conclude that the novel S1QELs are effective in vivo, and that site I_Q runs in vivo and is a significant driver of pathology in Sod2^-/- mice.

2016-04-01·Free Radical Biology and Medicine2区 · 医学

Regulation of endothelial dynamics by PGC-1α relies on ROS control of VEGF-A signaling

2区 · 医学

Article

作者: García-Quintans, Nieves ; Arza, Elvira ; Sánchez-Ramos, Cristina ; Luque, Alfonso ; Monsalve, María ; Olmos, Yolanda ; Prieto, Ignacio

Peroxisome proliferator activated receptor γ co-activator 1α (PGC-1α) is a regulator of mitochondrial metabolism and reactive oxygen species (ROS) that is known to play a relevant role in angiogenesis.AIMSThis study aims to investigate the role of ROS on the regulation by PGC-1α of angiogenesis.METHODS AND RESULTSWe found that endothelial cells (ECs) from mice deleted for PGC-1α display attenuated adhesion to the extracellular matrix, together with slower and reversible spreading. Structural analysis demonstrates unstable formation of focal adhesions, defective cytoskeleton reorganization in response to cellular matrix adhesion, cell migration and cell-cell adhesion. Confluent cultures showed also a reduction of membrane bound VE-cadherin, suggesting defective inter-cellular junction formation. Functional consequences included impaired directional migration, and enhanced tip phenotype in aortic explants sprouting assays. At the molecular level, PGC-1α-deleted ECs exhibit a constitutive activation of the vascular endothelial growth factor-A (VEGF-A) signaling pathway and a defective response to VEGF-A. All these alterations are partially reversed by administration of the antioxidant EUK-189. The contribution of mitochondrial ROS and NOX activation was confirmed using a mitochondrial targeted antioxidant (MitoTEMPO) and a NOX inhibitor (VAS-2870). These results indicate that elevated production of ROS in the absence of PGC-1α is a key factor in the alteration of the VEGF-A signaling pathway and the capacity of endothelial cells to form stable interactions with other endothelial cells and with the extracellular matrix. Our findings show that PGC-1α control of ROS homeostasis plays an important role in the control of endothelial response to VEGF-A.

2012-12-01·American Journal of Physiology-Lung Cellular and Molecular Physiology2区 · 医学

Antioxidant mimetics modulate oxidative stress and cellular signaling in airway epithelial cells infected with respiratory syncytial virus

2区 · 医学

Article

作者: Garofalo, Roberto P. ; Casola, Antonella ; Mautemps, Nicolas ; Komaravelli, Narayana ; Hosakote, Yashoda M. ; Liu, Tianshuang

Respiratory syncytial virus (RSV) is one of the most common causes of bronchiolitis and pneumonia among infants and young children worldwide. In previous investigations, we have shown that RSV infection induces rapid generation of reactive oxygen species (ROS), which modulate viral-induced cellular signaling, and downregulation of antioxidant enzyme (AOE) expression, resulting in oxidative stress in vitro and in vivo, which plays a pathogenetic role in RSV-induced lung disease. In this study, we determined whether pharmacological intervention with synthetic catalytic scavengers could reduce RSV-induced proinflammatory gene expression and oxidative cell damage in an in vitro model of infection. Treatment of airway epithelial cells (AECs) with the salen-manganese complexes EUK-8 or EUK-189, which possess superoxide dismutase, catalase, and glutathione peroxidase activity, strongly reduced RSV-induced ROS formation by increasing cellular AOE enzymatic activity and levels of the lipid peroxidation products F2-8-isoprostane and malondialdehyde, which are markers of oxidative stress. Treatment of AECs with AOE mimetics also significantly inhibited RSV-induced cytokine and chemokine secretion and activation of the transcription factors nuclear factor-κB and interferon regulatory factor-3, which orchestrate proinflammatory gene expression. Both EUKs were able to reduce viral replication, when used at high doses. These results suggest that increasing antioxidant cellular capacities can significantly impact RSV-associated oxidative cell damage and cellular signaling and could represent a novel therapeutic approach in modulating virus-induced lung disease.

100 项与 EUK-189 相关的药物交易

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