Astragalus polysaccharides inhibit oxidation in high glucose-challenged or SOD2-silenced H9C2 cells
Authors Chen W, Sun Q, Ju J, Chen W, Zhao X, Zhang Y, Yang Y
Received 14 June 2018
Accepted for publication 29 August 2018
Published 24 October 2018 Volume 2018:11 Pages 673—681
Checked for plagiarism Yes
Review by Single-blind
Peer reviewers approved by Ms Justinn Cochran
Peer reviewer comments 4
Editor who approved publication: Professor Ming-Hui Zou
Wei Chen,1 Qilin Sun,1 Jing Ju,1 Wenjie Chen,1 Xuelan Zhao,1 Yu Zhang,1 Yehong Yang2
1Department of Geriatrics, Huashan Hospital, Fudan University, Shanghai 200040, China; 2Department of Endocrinology, Huashan Hospital, Fudan University, Shanghai 200040, China
Introduction: Oxidative stress plays an important role in the development of diabetic cardiomyopathy (DCM). Previously, we reported that Astragalus polysaccharides (APS) improved DCM by inhibition of cardiac oxidative stress. In this study, we evaluated the beneficial effect of APS on high glucose-induced oxidative stress in cardiomyocytes in vitro.
Materials and methods: H9C2 cells were cultured in the presence of high concentration of glucose or transfected with siRNASOD2, followed by APS treatment. The cellular mitochondrial ultrastructure was observed using a transmission electron microscope. Cell apoptosis was detected using hairpin oligonucleotide probes and quantified by flow cytometry analysis. Superoxide production was determined by immunohistochemistry using the fluorescent dye dihydroethidium (DHE). Nitrotyrosine and 8-OH-dG antibodies were employed to detect oxidative damage to cytoplasmic proteins and oxidative stress in the nuclei, respectively. Superoxide dismutase (SOD) activity was measured utilizing the SOD Assay Kit, and SOD protein levels were analyzed by Western blotting.
Results: APS treatment protected cellular mitochondrial ultrastructure, reduced cell apoptosis (hairpin-1), inhibited cellular superoxide production (DHE), and reduced oxidative damage to cytoplasmic proteins (nitrotyrosine) and oxidative stress in the nuclei (8-OH-dG) in high glucose-induced and/or SOD2-silenced H9C2 cells, together with induction of SOD2 enzyme activity and increase of protein levels.
Conclusion: Our findings indicated the beneficial effect of APS on high glucose-challenged H9C2 cells, which was associated with inhibition of oxidative stress in vitro.
Keywords: superoxide dismutases, oxidative stress, apoptosis, cardiomyocyte, ROS, diabetic cardiomyopathy
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