title: ROS activates JNK-mediated autophagy to counteract apoptosis in mouse mesenchymal stem cells in vitro.
abstract: OBJECTIVE
  Transplantation of mesenchymal stem cells (MSCs) for the treatment of diabetic erectile dysfunction (ED) is hampered by apoptosis of the transplanted cells. In diabetic ED, there is increased oxidative stress and decreased NO in the corpora cavernosa, and reactive oxygen species (ROS) induce apoptosis of the transplanted cells. In this study we examined whether and how autophagy was involved in ROS-induced apoptosis of MSCs.
  
  
  METHODS
  Mouse C3H10 MSCs were treated with H2O2 to simulate the high oxidative condition in diabetic ED. Cell viability was measured using MTT assay. Apoptosis was analyzed by flow cytometry. Apoptosis- and autophagy-related proteins were detected with Western blot assays. Intracellular autophagosome accumulation was studied using transmission electron microscopy.
  
  
  RESULTS
  Treatment of MSCs with H2O2 (50-400 μmol/L) inhibited the cell viability in concentration- and time-dependent manners. Furthermore, H2O2 (300 μmol/L) induced apoptosis, as well as activated autophagy in MSCs. Pretreatment with lysosome inhibitor chloroquine (10 μmol/L) or PI3K inhibitor 3-methyladenine (5 mmol/L) significantly enhanced H2O2-induced cell death. Pretreatment with JNK inhibitor SP600125 (10 μmol/L) abrogated H2O2-induced accumulation of LC3-II, and attenuated H2O2-induced reduction of Bcl-2 levels in MSCs.
  
  
  CONCLUSIONS
  ROS induce autophagy to counteract apoptosis in MSCs by activation of JNK. Thus, augmentation of autophagy may reduce apoptosis, prolonging MSC survival and improving MSC-based therapeutic efficacy for diabetic ED.
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.|Chongqing Key Laboratory of Molecular Oncology and Epigenetics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Guan-yu
authors.initials: GY
authors.lastname: Liu
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xiao-xue
authors.initials: XX
authors.lastname: Jiang
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Wei-yang
authors.initials: WY
authors.lastname: He
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: You-lin
authors.initials: YL
authors.lastname: Kuang
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Ren
authors.affiliation: Molecular Biology and Lung Cancer Program, Lovelace Respiratory Research Institute, Albuquerque, NM 87108, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Gou
mesh_terms: D000818:Animals; D017209:Apoptosis; D001343:Autophagy; D002460:Cell Line; D002470:Cell Survival; D004789:Enzyme Activation; D007172:Erectile Dysfunction / Q000628:therapy; D006861:Hydrogen Peroxide / Q000378:metabolism; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D048670:MAP Kinase Kinase 4