大气细颗粒物对SH-SY5Y细胞能量代谢的影响
Effect of airborne fine particulate matters on energy metabolism of SH-SY5Y cells
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摘要:
目的 研究大气细颗粒物(PM2.5)对人神经母细胞瘤(SH-SY5Y)细胞能量代谢的影响。
方法 用不同质量浓度(下称浓度)PM2.5(0、20、40、80、160、320 mg/L)染毒SH-SY5Y细胞24 h, MTT法检测细胞存活率, 丙二醛(MDA)试剂盒检测MDA的含量, XFp细胞能量代谢分析仪检测细胞线粒体呼吸功能及糖酵解功能。
结果 PM2.5各浓度组SH-SY5Y细胞存活率均低于对照组(均P < 0.05); 80、160、320 mg/L PM2.5组细胞内MDA含量高于对照组(均P < 0.05)。PM2.5各浓度组细胞基础耗氧率、ATP偶联的有氧呼吸速率低于对照组(P < 0.05), 分别下降了13.8%、19.7%、25.1%、35.8%、45.2%和17.0%、21.9%、28.3%、39.2%、47.9%; 40、80、160、320 mg/L PM2.5组细胞有氧呼吸最大值较对照组下降了19.0%、24.2%、28.5%、40.7%(均P < 0.05)。80、160、320 mg/L PM2.5组细胞糖酵解水平和糖酵解最大值低于对照组(均P < 0.05), 分别下降了15.9%、22.1%、26.1%和16.5%、19.6%、21.5%。SH-SY5Y细胞基础耗氧率、ATP偶联的有氧呼吸速率、质子漏耗氧速率、有氧呼吸最大值、糖酵解水平及糖酵解最大值与细胞存活率呈正相关(r在0.65~0.86之间, P < 0.01), 与MDA值呈负相关(r为-0.53~-0.86, P < 0.05或P < 0.01)。
结论 PM2.5可引起SH-SY5Y细胞的氧化损伤, 降低细胞线粒体呼吸功能及糖酵解功能。
Abstract:Objective To study the effect of atmospheric fine particulate matters (PM2.5) on the energy metabolism of human neuroblastoma (SH-SY5Y) cells.
Methods SH-SY5Y cells were exposed to different concentrations of PM2.5 (0, 20, 40, 80, 160, 320 mg/L) for 24 h. Cell survival rate was detected by MTT assay, and malondialdehyde (MDA) content with MDA kit, and mitochondrial respiratory function and glycolytic function with XFp extracellular flux analyzer.
Results The survival rates of SH-SY5Y cells of the PM2.5 groups were lower than that of the control group (Ps < 0.05). The MDA contents of the 80, 160, and 320 mg/L PM2.5 groups were higher than that of the control group (Ps < 0.05). The PM2.5 groups showed lower basal oxygen consumption rate (OCR) and ATP-linked OCR than the control group (OCR reduced by 13.8%, 19.7%, 25.1%, 35.8%, and 45.2%, respectively; ATP-linked OCR reduced by 17.0%, 21.9%, 28.3%, 39.2%, and 47.9%, respectively) (P < 0.05). The 40, 80, 160, and 320 mg/L PM2.5 groups showed lower maximal respiration level than the control group (reduced by 19.0%, 24.2%, 28.5%, and 40.7%, respectively) (Ps < 0.05). The 80, 160, and 320 mg/L PM2.5 groups displayed lower glycolysis and maximum glycolysis than the control group (glycolysis reduced by 15.9%, 22.1%, and 26.1%, respectively; maximum glycolysis reduced by 16.5%, 19.6%, and 21.5%, respectively) (Ps < 0.05). Basal OCR, ATP-linked OCR, proton leak OCR, maximal respiration capacity, glycolysis level, and maximum glycolysis were positively associated with cell survival rate (r ranged from 0.65 to 0.86, P < 0.01), and negatively associated with MDA (r ranged from -0.53 to -0.86, P < 0.05 or P < 0.01).
Conclusion PM2.5 could cause oxidative damage to SH-SY5Y cells with decreasing the mitochondrial respiratory function and glycolysis function.
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