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27 June 2014 : Original article  

Resveratrol protects vascular smooth muscle cells against high glucose-induced oxidative stress and cell proliferation in vitro

Rong GuoABE, Weiming LiA, Baoxin LiuBF, Shuang LiBF, Buchun ZhangCDG, Yawei XuDG

DOI: 10.12659/MSMBR.890858

Med Sci Monit Basic Res 2014; 20:82-92

Abstract

BACKGROUND: Resveratrol exhibits beneficial effects against numerous degenerative diseases at different stages of pathogenesis. This study investigated potential mechanisms and resveratrol effects on high glucose (HG)-induced oxidative stress (30 mM d-glucose, 30 min) and cell proliferation (30 mM d-glucose, 24 h) in vascular smooth muscle cells (VSMCs).

MATERIAL AND METHODS: Intracellular reactive oxygen species (ROS) generation was detected by 2’,7’-dichlorofluorescein diacetate (DCFH-DA). Total antioxidant capacity (TAC), malonyldialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) were measured to evaluate oxidative stress. VSMC proliferation was measured by CCK-8 assays and through propidium iodide-based cell cycle analysis. Expression of NAD(P)H oxidase, proliferation proteins, and cell signalling were assessed by immunoblot analysis.

RESULTS: Co-treatment of primary cultures of VSMCs with 1–100 μM resveratrol decreased HG-induced ROS overproduction (P<0.05). Resveratrol also abolished HG-induced phosphorylation of oxidase subunit p47 phox and reduced HG-induced cyclin D1, cyclin E, and PCNA expression in a concentration-dependent manner. Furthermore, resveratrol (10 μM) attenuated HG-induced phosphorylation of Akt, p38 mitogen-activated protein kinase (MAPK), ERK 1/2, and JNK1/2 without affecting total levels. HG stimulation enhanced downstream IκB-α phosphorylation and NF-κB activity, and resveratrol repressed these effects.

CONCLUSIONS: Resveratrol inhibits HG-induced oxidative stress and VSMC proliferation by suppressing ROS generation, NADPH oxidase, Akt phosphorylation, p38 MAPK/JNK/ERK phosphorylation, and IκB-α and NF-κB activities.

Keywords: Biological Markers - metabolism, Animals, Cell Cycle - drug effects, Cell Proliferation - drug effects, Cell Survival - drug effects, Cells, Cultured, Cytoprotection - drug effects, Extracellular Signal-Regulated MAP Kinases - metabolism, Glucose - toxicity, I-kappa B Proteins - metabolism, JNK Mitogen-Activated Protein Kinases - metabolism, Models, Biological, Muscle, Smooth, Vascular - pathology, Myocytes, Smooth Muscle - pathology, NADPH Oxidase - metabolism, Oxidation-Reduction - drug effects, Oxidative Stress - drug effects, Phosphorylation - drug effects, Protective Agents - pharmacology, Proto-Oncogene Proteins c-akt - metabolism, Rats, Sprague-Dawley, Reactive Oxygen Species - metabolism, Signal Transduction - drug effects, Stilbenes - pharmacology, p38 Mitogen-Activated Protein Kinases

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Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416