title: SIRT3 mediates decrease of oxidative damage and prevention of ageing in porcine fetal fibroblasts.
abstract: OBJECTIVE
  Sirtuin 3 (SIRT3) is a mitochondria-specific protein required for the deacetylation of metabolic enzymes and the action of oxidative phosphorylation by acting as a nicotinamide adenine dinucleotide (NAD+)-dependent deacetylase. SIRT3 increases oxidative stress resistance and prevents mitochondrial decay associated with ageing in response to caloric restriction. However, the effects of SIRT3 on oxidative damage and ageing are not well understood. We investigated the physiological functions of porcine SIRT3 on the damage and ageing in porcine fetal fibroblasts (PFFs).
  
  
  METHODS
  Overexpression and knockdown of SIRT3 were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis, respectively. All cells were treated with three different stress reagents 12-o-tetradecanoylphorbol-13-acetate (TPA), methanesulfonic acid methylester (MMS), and tert-butylhydroperoxide (t-BHP), respectively, and then examined by flow cytometry following JC-1 (5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimidazol-carbocyanine iodide) staining.
  
  
  RESULTS
  SIRT3 overexpression enhanced the ability of superoxide dismutase 2 (SOD2) to reduce cellular reactive oxygen species (ROS), which further decreased the damage to the membranes and the organelles of the cells, especially to mitochondria. It inhibited the initial decrease of mitochondrial membrane potential, and prevented the decrease of adenosine triphosphate (ATP) production and activity of Nampt. In contrast, SIRT3 knockdown reduced the ability of SOD2 to increase cellular ROS which was directly correlated with stress-induced oxidative damage and ageing in PFFs.
  
  
  CONCLUSIONS
  Our findings identify one function of SIRT3 in PFFs was to dampen cytotoxicity, and, therefore, to decrease oxidative damage and attenuate ageing possibly by enhancing the activity of SOD2.
authors.affiliation: College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310032, China. Electronic address: xxxie@zjut.edu.cn.
authors.forename: Xiaoxian
authors.initials: X
authors.lastname: Xie
authors.affiliation: College of Ecology, Lishui University, Lishui 323000, China; College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Binggong
authors.initials: B
authors.lastname: Zhao
authors.affiliation: College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Li
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000375:Aging / Q000502:physiology*; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D015153:Blotting, Western; D002232:Carbocyanines / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D053078:Membrane Potential, Mitochondrial; D018384:Oxidative Stress / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D056566:Sirtuin 3 / Q000235:genetics / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism; D013552:Swine; D020122:tert-Butylhydroperoxide / Q000494:pharmacology; D000099156:Superoxide Dismutase 2
chemical_list: D000255:Adenosine Triphosphate; D001562:Benzimidazoles; D002232:Carbocyanines; D017382:Reactive Oxygen Species; D056566:Sirtuin 3; D013482:Superoxide Dismutase; D020122:tert-Butylhydroperoxide; D000099156:Superoxide Dismutase 2; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine
keywords: Ageing; Knockdown; Overexpression; Oxidative damage; Porcine SIRT3