ROS activates JNK-mediated autophagy to counteract apoptosis in mouse mesenchymal stem cells in vitro

Acta Pharmacol Sin. 2015 Dec;36(12):1473-9. doi: 10.1038/aps.2015.101. Epub 2015 Nov 23.

Abstract

Aim: 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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Autophagy*
  • Cell Line
  • Cell Survival
  • Enzyme Activation
  • Erectile Dysfunction / therapy
  • Hydrogen Peroxide / metabolism
  • MAP Kinase Kinase 4 / metabolism*
  • Male
  • Mesenchymal Stem Cell Transplantation
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Oxidative Stress
  • Reactive Oxygen Species / metabolism*

Substances

  • Reactive Oxygen Species
  • Hydrogen Peroxide
  • MAP Kinase Kinase 4