硫辛酸通过抑制NF-κB-iNOS-NO信号保护氧化应激诱导的PC12细胞损伤

Lipoic Acid Protected PC12 Cells from Oxidative Stress-induced Damage by inhibiting the NF-κB- iNOS-NO Signal Pathway

关键词:  硫辛酸  过氧化氢  PC12细胞  一氧化氮  iNOS mRNA  NF-κB p65 mRNA

Key words:  lipoic acid  H2O2  PC12 cells  NO  iNOS mRNA  NF-κB mRNA

摘要:

目的 研究硫辛酸对氧化应激的PC12细胞NF-κB - iNOS-NO信号通路的影响。方法 用终浓度为0.4 mmol·L-1的H2O2损伤PC12细胞，用MTT法测定细胞存活率；用激光共聚焦法(LSCM)检测细胞内一氧化氮的动态变化；用RT-PCR法检测iNOS mRNA和NF-κB p65 mRNA表达量的变化。结果 0.4 mmol·L-1的H2O2处理PC12细胞4 h，细胞存活率为27.9%(P<0.01)，硫辛酸10 mg·L-1可以提高H2O2损伤的PC12细胞存活率(65.4%，P<0.01)；LSCM检测显示，DAF-2荧光强度的比值(Ft/F0)的最大值由损伤模型组的5.34下降到1.80；和模型组比较，iNOS mRNA和NF-κB p65 mRNA的表达明显下调 (P<0.05或P<0.01)。结论 硫辛酸可能通过抑制NF-κB - iNOS-NO信号减轻氧化应激诱导的PC12细胞损伤。

Abstract:

OBJECTIVE To investigate the effect of lipoic acid(LA) on NF-κB - iNOS-NO signal pathway in oxidative stress-induced PC12 cells. METHODS After PC12 cells were treated with 0.4 mmol·L-1 H2O2 and different concentrations of LA, the cell viability was measured by MTT, the concentration of nitric oxide in intracellular cells was evaluated using laser-scanning confocal microscopy(LSCM) method and the expressions of NF-κB p65, iNOS mRNA were detected by RT-PCR. RESULTS Cell viability were significantly elevated by LA from 27.9% to 65.4%. Compared with H2O2-treated cells, the concentration of nitric oxide was lowed by LA through judging the change of DAF-2 Ft/F0 from 5.34 to 1.80. Meanwhile, the levels of NF-κB p65 and iNOS mRNA were decreased (P<0.05 or P<0.01). CONCLUSION Lipoic acid could protect PC12 cells from oxidative stress-induced damage by inhibiting the NF-κB - iNOS-NO signal pathway.