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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 investigate the effect of di-2-ethylhexyl phthalate (DEHP) on the apoptosis of mouse spermatocytes (GC-2 spd) and the expression of TR4 mRNA. Methods GC-2 spd cells were exposed to medium containing 0 (solvent control) and 50, 100 and 200 μmol / L DEHP for 24 h. The cell viability was detected by MTT assay. The relative expression level of TR4 mRNA was detected by Real-time PCR and the apoptosis was detected by flow cytometry. Results Compared with the solvent control group, the survival rate of GC-2 spd cells in 200 μmol / L DEHP exposure group was low, and the apoptosis rate of GC-2 spd cells in 100 and 200 μmol / L DEHP exposure group was high, only 200 μmol / L DEHP exposure group GC-2 spd cells TR4 mRNA expression levels were higher, the difference was statistically significant (P <0.05); and with the dose of DEHP increased, the survival rate of GC-2 spd cells Showed a downward trend, the rate of apoptosis showed an upward trend. Conclusion DEHP has reproductive toxicity on spermatogenic cells, which can induce the apoptosis of spermatogenic cells by promoting orphan receptor TR4.