Abstract: OBJECTIVE: To investigate the effects of di(2-ethylhexyl) phthalate (DEHP) and its active metabolite, mono(2-ethylhexyl) phthalate (MEHP) on ovarian granulosa cells. METHODS: ICR mice ages at the 21 days were used by subcutaneous injection PMSG. The ovarian granulosa cells were collected and cultured in DMEM-F12. After the ovarian granulosa cells were prepared, cells were exposed to doses of 10, 50, 250 nmol/L DEHP and 10, 50, 250 nmol/L MEHP for 24 h. The mRNA levels of StAR, P450scc were determined by RT-PCR. The mRNA levels of CYP19, PPARα, PPARβ, PPARγ were determined by Real time RT-PCR. Estradiol and progesterone were measured by enzyme immunoassay (EIA) in ovarian granulosa cell. RESULTS: 250 nmol/L MEHP group decreased the gene expression of StAR and P450scc (P<0.05); 250 nmol/L DEHP group increased the gene expression of CYP19, but 10, 50 nmol/L DEHP and 50, 250 nmol/L MEHP groups decreased the gene expression of CYP19 (P<0.05). Except 10 nmol/L DEHP group, DEHP and MEHP decreased the gene expression of PPARα and PPARβ (P<0.05). 10 nmol/L DEHP and MEHP groups increased the gene expression of PPARγ, but 250 nmol/L MEHP group decreased the gene expression of PPARγ (P<0.05). DEHP and MEHP enhanced the concentration of estradiol at all dose groups, but 10 nmol/L DEHP, 50 and 250 nmol/L MEHP groups decreased the concentration of progesterone in ovarian granulosa cells (P<0.05). CONCLUSION: DEHP and MEHP could affect the gene expression of steroid biosynthesis enzymes, PPARs, and the secretory function in ovarian granulosa cells.

Key words: reproductive toxicity, di(2-ethylhexyl) phthalate, mono(2-ethylhexyl) phthalate, ovarian granulosa cells