邻苯二甲酸二(2-乙基己基)酯对小鼠精母细胞凋亡及孤核受体TR4 mRNA表达的影响

Effects of diethyl hexyl phthalate on apoptosis and expression of TR4 mRNA in GC-2 spd cells

目的研究邻苯二甲酸二(2-乙基己基)酯(di-2-ethylhexyl phthalate,DEHP)对小鼠精母细胞(GC-2 spd)凋亡及TR4 mRNA表达的影响。方法将GC-2 spd细胞分别暴露于含终浓度0(溶剂对照)、50、100、200μmol/L DEHP的培养基培养24 h。采用MTT法检测细胞活性,采用Real-time PCR方法检测细胞TR4 mRNA的相对表达水平,流式细胞仪检测细胞凋亡情况。结果与溶剂对照组比较,200μmol/L DEHP染毒组GC-2 spd细胞的存活率均较低,100、200μmol/L DEHP染毒组GC-2 spd细胞的凋亡率均较高,仅200μmol/L DEHP染毒组GC-2 spd细胞TR4 mRNA的表达水平较高,差异均有统计学意义(P<0.05);且随着DEHP染毒剂量的升高,GC-2 spd细胞的存活率呈下降趋势,凋亡率呈上升趋势。结论 DEHP对生精细胞具有生殖毒性作用,可通过促进孤核受体TR4诱导生精细胞凋亡。

Objective To study the effects of di-（2-ethylhexyl） phthalate （DEHP） on the GC-2 spd cell apoptosis and the expression of TR4 mRNA. Methods GC-2 spd cells were respectively exposed to the medium with the final concentrations of 0,50,100,200μmol/L （with solvent as the control） for DEHP and then cultured for 24 h. Cell activity was detected by MTF, the relative expression level of TR4 mRNA was detected by real-time PCR method, and cell apoptosis was detected by flow cytometry. Results Compared with the solvent control group,the survival rate of GC-2 spd cells in the 200 I^mol/L DEHP exposure group was lower,the apoptosis rates of GC-2 spd cells in the 100 and 200 p^mol/L DEHP exposure groups were higher,and only the expression level of TR4 mRNA of GC-2 spd cells was higher,with statistically significant differences （P〈0.05）. With the increase of the DEHP dose,the survival rate of GC-2 spd cells showed a decreasing trend,while the apoptosis rate showed an increasing trend. Conclusion DEHP has a reproductive toxic effect on spermatogenic cells,which can induce the apoptosis of spermatogenic cells by the orphan nuclear receptor 4.