The protective mechanism of dl-3-n butylphthalide on oxidative stress injury in rat bone nmrmw mesenchynml stem cells

Abstract

Objective To study the protective mechanism of dl-3-n butylphthalide （NBP） on oxidative stress injury induced by hydrogea peroxide （H2O2） in rat bone marrow stem cells （rBMSCs）. Methods The rBMSCs were divided into control, HE O2 and different concentration NBP pretreatment groups. The control group received no treatment. An oxidative stress injury model was induced by H2O2 for 4 hours with the final concentration 600 μmol/L in the H2O2 group. In the NBP pretreatment groups, the rBMSCs were pretreated with different NBP concentrations （0. 1, 1, 10, and 100μmol/L） for 24 hours, then treated with H2O2 for 4 hours with the final concentration 600μ mol/L. The cell viability was detected by MTT method. The apoptosis rate was detected by flow cytometry. Superoxide dismutase （SOD） activity and malondialdehyde （MDA） content were measured with SOD and MDA commercial kits. Results The cell activity （A） was 0. 487±0. 018 in the H2O2 group, and it was significantly lower than 0. 750±0. 016 in the control group （P =0. 000）; they were 0. 597±0. 024, 0. 666±0. 033, and 0. 658 ±0. 012 in the NBP 1, 10, and 100μ mol/L pretreatment groups, they were all signifi- cantly higher than the H2O2 group （all P =0. 000）, and showed a dose dependent manner. The apoptosis rate was （44. 96 ± 2. 84）% in the H2O2 group, and it was significantly higher than （0. 15 ±0.07）% in the control group （P= 0.000）. The apoptosis rates were （31.79±1.60）% ,（21.41± 1.92）% and （22.59± 1.78）% in the NBP 1, 10, and 100 μmol/L pretreat- merit groups, they were all significantly lower than the H20, groups （all P = 0. 000）, and showed a dose-dependent manner. The SOD activity was （24. 01 ± 2. 85） U/mg in the H2O2 group （P = 0. 000）, and it was significantly lower than （43.58 ±2. 72） U/mg in the control group （P =0. 000）; they were （28. 29±1.19）, （34. 06± 1.83）, and （31.76 ± 1.75） U/mg in the NBP 1, 10, and 100 μmol/L pretreatment groups, and they were all significantly higher than the H2O2 group （all P = 0. 000）. The MDA content was （7. 98 ±0. 55） nmol/mg in the H2O2 group, and it was significantly higher than （4. 73±0. 53） nmol/mg in the control group （P = 0. 000）; they were （6. 97 ±0. 29）, 6. 09 ±0. 28）, and （6. 15±0. 41） nmol/mg respectively in the NBP 1, 10, and 100μmol/L pretreatment groups （P = 0. 000）, they were significantly lower than the H2O2 group, and showed a dose-dependent rammer. Conclusions NBP has obvious protective effects on oxidative stress injury induced by H2O2 in rBMSCs. Its mechanism may be associated with the role of antioxidant oxidative stress of NBP. Key words: 3-n-Butylphthalide; Oxidative stress; Bone marrow cells; Mesenchymal stem cells; Neuroprotective agents; Antioxidants; Rats