Chondroitin sulfate protects SH-SY5Y cells from oxidative stress by inducing heme oxygenase-1 via phosphatidylinositol 3-kinase/Akt

J Pharmacol Exp Ther. 2007 Dec;323(3):946-53. doi: 10.1124/jpet.107.123505. Epub 2007 Sep 20.

Abstract

We investigated the mechanism of the neuroprotective properties of chondroitin sulfate (CS), an endogenous perineuronal net glycosaminoglycan, in human neuroblastoma SH-SY5Y cells subjected to oxidative stress. Preincubation with CS for 24 h afforded concentration-dependent protection against H2O2-induced toxicity (50 microM for 24 h) measured as lactic dehydrogenase released to the incubation media; cell death was prevented at the concentrations of 600 and 1000 microM. Cell death caused by a combination of 10 microM rotenone plus 1 microM oligomycin-A (Rot/oligo) was also reduced by CS at concentrations ranging from 0.3 to 100 microM; in this toxicity model, maximum protection was achieved at 3 microM (48%). No significant protection was observed in a cell death model of Ca2+ overload (70 mM K+, for 24 h). H2O2 and Rot/oligo generated reactive oxygen species (ROS) measured as an increase in the fluorescence of dichlorofluorescein diacetate-loaded cells. CS drastically reduced ROS generation induced by both H2O2 (extracellular ROS) and Rot/oligo (intracellular ROS). CS also increased the expression of phosphorylated Akt and heme oxygenase-1 by 2-fold. The protective effects of CS were prevented by chelerythrine, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), cycloheximide, and Sn(IV)-protoporphyrin IX. Taken together, these results show that CS can protect SH-SY5Y cells under oxidative stress conditions by activating protein kinase C, which phosphorylates Akt that, via the phosphatidylinositol 3-kinase/Akt pathway, induces the synthesis of the antioxidant protein heme oxygenase-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Chondroitin Sulfates / pharmacology*
  • Enzyme Induction
  • Flow Cytometry
  • Heme Oxygenase-1 / biosynthesis*
  • Humans
  • Hydrogen Peroxide / toxicity
  • Immunoblotting
  • L-Lactate Dehydrogenase / metabolism
  • Neuroprotective Agents / pharmacology*
  • Oligomycins / toxicity
  • Oxidants / toxicity
  • Oxidative Stress / drug effects*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Reactive Oxygen Species / metabolism
  • Rotenone / toxicity

Substances

  • Neuroprotective Agents
  • Oligomycins
  • Oxidants
  • Reactive Oxygen Species
  • Rotenone
  • oligomycin A
  • Chondroitin Sulfates
  • Hydrogen Peroxide
  • L-Lactate Dehydrogenase
  • Heme Oxygenase-1
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt