title: Advanced glycation end products promote the proliferation and migration of primary rat vascular smooth muscle cells via the upregulation of BAG3.
abstract: The present study was aimed to investigate the role of reactive oxygen species (ROS) on advanced glycation end product (AGE)-induced proliferation and migration of vascular smooth muscle cells (VSMCs) and whether Bcl-2‑associated athanogene 3 (BAG3) is involved in the process. Primary rat VSMCs were extracted and cultured in vitro. Cell viability was detected by MTT assay and cell proliferation was detected by EdU incorporation assay. Cell migration was detected by wound healing and Transwell assays. BAG3 was detected using qPCR and western blot analysis. Transcriptional and translational inhibitors (actinomycin D and cycloheximide, respectively) were used to study the effect of AGEs on the expression of BAG3 in VSMCs. Lentiviral plasmids containing short hairpin RNA (shRNA) against rat BAG3 or control shRNA were transduced into VSMCs. Cellular ROS were detected by 2',7'-dichlorofluorescein diacetate (DCFH-DA) staining. Mitochondrial membrane potential was detected by tetramethylrhodamine methyl ester (TMRE) staining. AGEs significantly increased the expression of BAG3 in a dose-and time-dependent manner. Furthermore, AGEs mainly increased the expression of BAG3 mRNA by increasing the RNA synthesis rather than inhibiting the RNA translation. BAG3 knockdown reduced the proliferation and migration of VSMCs induced by AGEs. BAG3 knockdown reduced the generation of ROS and sustained the mitochondrial membrane potential of VSMCs. Reduction of ROS production by N-acetylcysteine (NAC), a potent antioxidant, also reduced the proliferation and migration of VSMCs. On the whole, the present study demonstrated for the first time that AGEs could increase ROS production and promote the proliferation and migration of VSMCs by upregulating BAG3 expression. This study indicated that BAG3 should be considered as a potential target for the prevention and/or treatment of vascular complications of diabetes.
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Cunshu
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Yintao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Ning
authors.initials: N
authors.lastname: Ye
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Dongxue
authors.initials: D
authors.lastname: Dai
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors.forename: Yingxian
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D055785:Gene Knockdown Techniques; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D017127:Glycation End Products, Advanced; D017382:Reactive Oxygen Species