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目的探讨邻苯二甲酸二(2-乙基己)酯(di-2-ethylhexyl phthalate,DEHP)对GC-2spd细胞Bcl-2、Bax m RNA表达的影响。方法体外培养GC-2spd细胞,用不同浓度的DEHP(0、50、100和200μmol/L)染毒24 h。采用MTT实验检测细胞存活率,Annexin V-FITC/PI双染色法结合流式细胞仪检测细胞凋亡率,荧光定量PCR法检测Bcl-2和Bax的m RNA表达。结果在DEHP染毒细胞24 h后,随着DEHP浓度(0、50、100和200μmol/L)的增加,细胞存活率逐渐下降,细胞凋亡率逐渐上升,Bcl-2 m RNA表达水平下降,Bax m RNA表达水平上升。结论 DEHP可能通过线粒体通路诱导GC-2spd细胞凋亡,进而影响雄性生殖功能。
Objective To investigate the effect of di-2-ethylhexyl phthalate (DEHP) on the expression of Bcl-2 and Bax m RNA in GC-2 spd cells. Methods GC-2spd cells were cultured in vitro and exposed to various concentrations of DEHP (0, 50, 100 and 200 μmol / L) for 24 h. Cell viability was detected by MTT assay. Apoptosis rate was detected by Annexin V-FITC / PI double staining combined with flow cytometry. The mRNA expression of Bcl-2 and Bax was detected by real-time quantitative PCR. RESULTS: After treated with DEHP for 24 h, the cell viability decreased gradually with the increase of DEHP concentration (0, 50, 100 and 200 μmol / L), the apoptosis rate gradually increased and the expression level of Bcl-2 mRNA decreased. Bax m RNA expression increased. Conclusion DEHP may induce apoptosis of GC-2 spd cells through mitochondrial pathway, which may affect male reproductive function.