邻苯二甲酸二乙基己酯对小鼠初级精母细胞氧化应激及凋亡的影响

Effects of Di-2-ethylhexyl Phthalate on Oxidative Stress and Apoptosis in GC-2 spd Cells

目的研究邻苯二甲酸二乙基己酯(DEHP)对小鼠初级精母细胞(GC-2 spd)氧化应激及凋亡的影响。方法体外传代培养永生化的GC-2 spd细胞,用二甲基亚砜(DMSO)溶解DEHP,以DMSO浓度为0.1%的细胞孔作为对照组,用DEHP终浓度为50、100和200μmol/L的细胞孔作为低、中、高剂量组染毒24 h。分光光度计法测定GC-2 spd细胞丙二醛(MDA)含量,超氧化物歧化酶(SOD)活力和谷胱甘肽过氧化物酶(GSH-Px)活力,流式细胞仪检测不同剂量组细胞凋亡情况。结果与对照组比较,DEHP低、中、高剂量染毒组MDA含量依次升高,差异无明显统计学意义(P>0.05);SOD活力在DEHP高剂量染毒组下降,差异有统计学意义(P<0.05);DEHP低、中剂量染毒组GSH-Px活力明显下降,且差异有统计学意义(P<0.05)。DEHP中、高剂量染毒组细胞凋亡明显高于对照组,且差异有统计学意义(P<0.05)。结论 DEHP可通过诱发小鼠生精细胞氧化应激而诱导细胞凋亡。

Objective To explore the effect of di-2-ethylhexyl phthalate（ DEHP） on oxidative stress and apoptosis in GC-2 spd cells. Methods Immortalized GC-2 spd cells were subcultured and treated with different concentration of DEHP（ 0,50,100 and 200 μmol / L） for 24 h; DEHP was dissolved in 0. 1% dimethyl sulfoxide（ DMSO）. The content of MDA,the activity of superoxide dismutase（ SOD） and glutathione peroxidase（ GSH-Px） in GC-2 spd cells were detected by spectrophotometry,and the apoptosis of GC-2 spd cells was analyzed by flow cytometry. Results Compared with the control group（ 0 μmol / L DEHP）,the content of MDA was increased with the increase of DEHP,but the difference was not statistically significant（ P〉 0. 05）. The activity of SOD was decreased significantly in the 200 μmol / L DEHP group（ P〈 0. 05）,and the activity of GSH-Px was decreased in the 50 μmol / L and 100 μmol / L DEHP group. The apoptosis of GC-2 spd cells was increased significantly in 100 μmol / L and200 μmol / L DEHP group（ P〈 0. 05）. Conclusion DEHP might induce apoptosis in GC-2 spd cells by oxidative stress.