Sargassum horneri methanol extract rescues C2C12 murine skeletal muscle cells from oxidative stress-induced cytotoxicity through Nrf2-mediated upregulation of heme oxygenase-1

Abstract

BackgroundSargassum horneri, an edible marine brown alga, is typically distributed along the coastal seas of Korea and Japan. Although several studies have demonstrated the anti-oxidative activity of this alga, the regulatory mechanisms have not yet been defined. The aim of the present study was to examine the cytoprotective effects of S. horneri against oxidative stress-induced cell damage in C2C12 myoblasts.MethodsWe demonstrated the anti-oxidative effects of a methanol extract of S. horneri (SHME) in a hydrogen peroxide (H2O2)-stimulated C2C12 myoblast model. Cytotoxicity was determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyl-tetrazolium assay and mode of cell death by cell cycle analysis. DNA damage was measured using a comet assay and expression of phospho-histone γH2A.X (p-γH2A.X). Levels of cellular oxidative stress as reactive oxygen species (ROS) accumulation were measured using 2’,7’-dichlorofluorescein diacetate. The involvement of selected genes in the oxidative stress-mediated signaling pathway was explored using Western blot analysis.ResultsSHME attenuated H2O2-induced growth inhibition and exhibited scavenging activity against intracellular ROS that were induced by H2O2. The SHME also inhibited comet tail formation, p-γH2A.X expression, and the number of sub-G1 hypodiploid cells, suggesting that it prevents H2O2-induced cellular DNA damage and apoptotic cell death. Furthermore, the SHME significantly enhanced the expression of heme oxygenase-1 (HO-1) associated with induction of nuclear factor-erythroid 2 related factor 2 (Nrf2) in a time- and concentration-dependent manner. Moreover, the protective effect of the SHME on H2O2-induced C2C12 cell damage was significantly abolished by zinc protoporphyrin IX, a HO-1 competitive inhibitor, in C2C12 cells.ConclusionsThese findings suggest that the SHME augments cellular antioxidant defense capacity through both intrinsic free radical scavenging activity and activation of the Nrf2/HO-1 pathway, protecting C2C12 cells from H2O2-induced oxidative cytotoxicity.

Adiponectin modulates oxidative stress-induced mitophagy and protects C2C12 myoblasts against apoptosis

“…As previously reported, ROS levels increased in H 2 O 2 -treated cells compared with untreated cells 37 . However, ROS levels were significantly inhibited in the presence of APN (Fig.…”

“…We next investigated whether APN affected intracellular ROS generation due to H 2 O 2 treatment using a 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. As previously reported, ROS levels increased in H 2 O 2 -treated cells compared with untreated cells 37 . However, ROS levels were significantly inhibited in the presence of APN (Fig.…”

Ameliorative role of curcumin on copper oxide nanoparticles‐mediated renal toxicity in rats: An investigation of molecular mechanisms

“…Curcumin γ-GCS. [63][64][65] Accordingly, transcription of γ-GCS [66,67] and HO-1 [68,69] is regulated by ROS-dependent signaling through activation of Nrf2.…”

The role of oxidative stress in skeletal muscle injury and regeneration: focus on antioxidant enzymes

“…In response to oxidative stress, Keap1 is oxidized, and dissociates from Nrf2, which in turn translocates to nucleus, heterodimerizes with Maf proteins, and binds to ARE, to induce expression of ARE-dependant genes (Ma 2013 ; Powers et al 2011 ; Trachootham et al 2008 ). In this manner, transcription of both GCS (Chan and Kwong 2000 ; Ding et al 2008 ) and HO-1 (Kang et al 2015 ; Sun et al 2015 ) is regulated by ROS-dependent signalling in myogenic cells.…”