二甲基甲酰胺致心肌细胞损伤及维生素C保护作用的实验研究

Study on the dimethylformamide-induced damage to cardiomyocytes and the protective effect of vitamin C

目的探讨二甲基甲酰胺(dimethylformamide,DMF)致H9c2心肌细胞损伤及维生素C(vitamin C,VC)对此损伤是否有保护作用。方法观察DMF染毒及VC干预后心肌细胞及核形态改变;活性氧(reactive oxygen species,ROS)、总抗氧化能力(total antioxidant capacity,T-AOC)试剂盒检测不同浓度DMF染毒不同时间、特定浓度DMF合并不同浓度VC干预后细胞氧化应激水平变化。结果 DMF染毒后细胞及核形态出现不同程度异常,低浓度VC干预后有所改善;100、140、180、220、250 m M DMF染毒24 h,100、140、180、200、220 m M DMF染毒48 h,100、125、140、180、200 m M DMF染毒72 h后,同时间不同剂量染毒,细胞ROS、T-AOC水平差异有统计学意义;同剂量不同时间染毒,细胞ROS、T-AOC水平差异有统计学意义(ROS:24 h:F=5 763.00,P<0.001;48 h:F=5 861.00,P<0.001;72 h:F=9 188.00,P<0.001;T-AOC:24 h:F=6.25,P=0.004;48 h:F=11.48,P<0.001;72 h:F=14.13,P<0.001)。VC(0.025、0.05、0.10、0.25 m M)干预后ROS、T-AOC水平与对照相比差异均有统计学意义(均有P<0.05)。结论氧化损伤可能是DMF发挥毒性的重要机制;低剂量VC可缓解DMF产生的氧化损伤,对细胞产生保护效应。

Objective To explore the damage of dimethylformamide( DMF) to H9c2 cardiomyocytes and a protective effect of vitamin C( VC).Methods The H9c2 cardiomyocytes were exposed to DMF with different concentrations,and then treated with different concentrations of VC to observe the protective effect of VC.The morphologic changes of cells were observed after DMF exposure and VC intervention.Reactive oxygen species( ROS) and total antioxidant capacity( TAOC) kits were used to detect oxidative stress levels treated with different concentrations of DMF and vitamin C for different time intervals.Results Abnormal morphologic changes of cells and nuclei were observed after exposure to DMF,while low concentrations of VC had a protective effect on the damage.After exposure of cardiomyocytes cells to the doses of 100,140,180,220,250 m M DMF treatment for 24 h,doses 100,140,180,200,220 m M DMF treatment for 48 h,and doses of 100,125,140,180,200 m M DMF treatment for 72 h,the results showed that ROS and T-AOC levels were significantly different under the same exposure time interval rather than different exposure doses,as well as under the same exposure dose rather than different exposure time intervals( R0S:24 h:F = 5 763.00,P < 0.001; 48 h:F = 5 861.00,P < 0.001; 72h:F = 9 188.00,P < 0.001; T-AOC:24 h:F = 6.25,P = 0.004; 48 h:F = 11.48,P < 0.001; 72 h:F = 14.13,P <0.001).After different doses of VC intervention( 0.025,0.05,0.10,0.25 m M),the ROS and T-AOC levels were significantly different compared with control group( P < 0.05).Conclusions Oxidative damage may be the mechanism in exerting the toxicity of DMF.Low-dose VC has a protective effect on oxidative damage of DMF to cardiomyocytes.