NecroX-5 protects mitochondrial oxidative phosphorylation capacity and preserves PGC1α expression levels during hypoxia/reoxygenation injury

Abstract

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.

NecroX-5 exerts anti-inflammatory and anti-fibrotic effects via modulation of the TNFα/Dcn/TGFβ1/Smad2 pathway in hypoxia/reoxygenation-treated rat hearts

“…Our previous proteomics study and network analysis [ 35 ] revealed a small protein network that comprised three identified proteins (Dcn, collagen alpha-2[I] chain [Col1a2], and lumican [Lum]) and two predicted networking proteins (TGFβ1 and SMAD2; Fig. 1B ).…”

“…LC-MS and a non-labeled peptide count protein quantification method were used to analyze the proteomes of rat hearts as described previously [ 35 36 ]. Heart proteins were separated via SDS-PAGE on a 12% polyacrylamide gel.…”

Mitochondria-Derived Vesicles Deliver Antimicrobial Reactive Oxygen Species to Control Phagosome-Localized Staphylococcus aureus

“…IRE1a deficiency suppressed the ability of MRSA-infected macrophages to induce mH 2 O 2 compared to the non-targeted control (Figure 1E). To test the requirement for mROS in killing MRSA, we treated infected macrophages with the ROS scavenger NecroX-5, which is primarily localized to mitochondria (Kim et al, 2010;Thu et al, 2016). Infected macrophages treated with NecroX-5 exhibited significantly lower MitoPY1 fluorescence than control-treated cells ( Figure 1F) and decreased capacity to kill MRSA ( Figure 1G).…”

Mitochondria-derived vesicles deliver antimicrobial payload to control phagosomal bacteria

“…1E). To test the requirement for mROS in killing MRSA, infected macrophages were treated with the ROS scavenger, NecroX-5, which is primarily localized to mitochondria (Kim et al, 2010; Thu et al, 2016). Infected macrophages treated with NecroX-5 exhibited lower MitoPY1 fluorescence than control-treated cells (Fig.…”