Protective Effects Of Glutathione On Hydrogen Peroxide-induced Cell Death And Matrix Destruction In Human Nucleus Pulposus Cells

Objective To investigate the protective effects of glutathione (GSH) on hydrogen peroxide (E2O2)-induced cell death and matrix destruction in human nucleus pulposus cells.Methods Human nucleus pulposus cells were cultured in alginate beads. Firstly, alginate beads were treated by different doses of H2O2. Prepare hydrogen peroxide-induced cell death and matrix destruction in cultured human NP cellmodel. Cells in alginate beads were treated as several designed groups:1, Untreated group; 2, H2O2 group; 3, GSH/H2O2 group. Viability of cells was assessed by LIVE/DEAD staining and apoptosis staining. The viability percentage was calculated by manually counting the number of dead/apoptosis cells and dividing. Real time PCR analysis of two extracellular matrix genes aggrecan and typeâ…¡collagen was performed.Results LIVE/DEAD Staining showed H2O2 at high concentration (500 uM) promotes the nucleus pulposus cell death (P<0.01). H2O2 at the concentrations of bigger than 4 uM decreased significantly gene expression of aggrecan and Type collagenâ…¡in NP cells (P<0.05). LIVE/DEAD staining showed in H2O2 group dead cell rate was higher than in GSH/H2O2 group (P<0.01). Apoptosis staining showed similar result (P<0.05). Gene expression of aggrecan and type collagenâ…¡in H2O2 group decreased more significantly than in GSH/H2O2 group (P<0.05).Conclusion Low concentration (20uM) of H2O2-induced matrix breakdown and high concentration (500uM) of H2O2-induced cell death can be efficiently prevented by 1 mM glutathione, implying that antioxidative reagents might be a novel treatment option for intervertebral disc degeneration.