title: SILAC-based proteomic analysis reveals that salidroside antagonizes cobalt chloride-induced hypoxic effects by restoring the tricarboxylic acid cycle in cardiomyocytes.
abstract: Hypoxic status alters the energy metabolism and induces cell injury in cardiomyocytes, and it further triggers the occurrence and development of cardiovascular diseases. Our previous studies have shown that salidroside (SAL) exhibits anti-hypoxic activity. However, the mechanisms remain obscure. In the present study, we successfully screened 92 different expression proteins in CoCl2-induced hypoxic conditions, 106 different expression proteins in the SAL-mediated anti-hypoxic group were compared with the hypoxic group using quantitative proteomics strategy, respectively. We confirmed that SAL showed a positive protective function involving the acetyl-CoA metabolic, tricarboxylic acid (TCA) cycle using bioinformatics analysis. We also demonstrated that SAL plays a critical role in restoring the TCA cycle and in protecting cardiomyocytes from oxidative injury via up-regulation expressions of PDHE1-B, ACO2, SUCLG1, SUCLG2 and down-regulation of MDH2. SAL also inhibited H9c2 cell apoptosis by inhibiting the activation of pro-apoptotic molecules caspase 3 and caspase 9 as well as activation of the anti-apoptotic molecular Bcl-2. Additionally, SAL also improved mitochondrial membrane potential (ΔΨm), reduced reactive oxygen species (ROS) and intercellular Ca(2+) concentration ([Ca(2+)]i) accumulation and inhibited the excessive consumption of ATP in H9c2 cells.
authors.affiliation: Central Laboratory, Logistics University of the Chinese People's Armed Police Force, 300309, China.
authors.forename: Zhong-Wei
authors.initials: ZW
authors.lastname: Xu
authors.affiliation: Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Tianjin 300309, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury, Tianjin 300162, China.
authors.forename: Xiao-Han
authors.initials: XH
authors.lastname: Jin
authors.affiliation: Department of Physiology and Pathophysiology, Logistics University of the Chinese People's Armed Police Force, Tianjin 300309, China.
authors.forename: Xiang-Yan
authors.initials: XY
authors.lastname: Meng
authors.affiliation: Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury, Tianjin 300162, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Central Laboratory, Logistics University of the Chinese People's Armed Police Force, 300309, China.
authors.forename: Feng-Xu
authors.initials: FX
authors.lastname: Fan
authors.affiliation: Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Tianjin 300309, China.
authors.forename: Shi-Yun
authors.initials: SY
authors.lastname: Mao
authors.affiliation: Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Tianjin 300309, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury, Tianjin 300162, China.
authors.forename: Wen-Cheng
authors.initials: WC
authors.lastname: Zhang
authors.affiliation: Central Laboratory, Logistics University of the Chinese People's Armed Police Force, 300309, China.
authors.forename: Na-Na
authors.initials: NN
authors.lastname: Shan
authors.affiliation: Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury, Tianjin 300162, China. Electronic address: cardiolab@live.com.
authors.forename: Yu-Ming
authors.initials: YM
authors.lastname: Li
authors.affiliation: Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Tianjin 300309, China. Electronic address: xu_rc@sohu.com.
authors.forename: Rui-Cheng
authors.initials: RC
authors.lastname: Xu
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D017209:Apoptosis; D002118:Calcium / Q000737:chemistry; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002460:Cell Line; D002853:Chromatography, Liquid; D002952:Citric Acid Cycle; D003035:Cobalt / Q000737:chemistry*; D019295:Computational Biology; D005960:Glucosides / Q000737:chemistry*; D000860:Hypoxia / Q000473:pathology; D008564:Membrane Potentials; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010100:Oxygen / Q000737:chemistry; D010636:Phenols / Q000737:chemistry*; D010936:Plant Extracts / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D029771:Rhodiola / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014233:Tricarboxylic Acids / Q000737:chemistry*
chemical_list: D005960:Glucosides; D010636:Phenols; D010936:Plant Extracts; D020543:Proteome; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D014233:Tricarboxylic Acids; D003035:Cobalt; D000255:Adenosine Triphosphate; D053148:Caspase 3; D053453:Caspase 9; C018021:cobaltous chloride; C009172:rhodioloside; D010100:Oxygen; D002118:Calcium
keywords: Cobalt chloride; Hypoxic; Quantitative proteomics; SILAC; Salidroside