Effects of Di-2-ethylhexyl Phthalate on the Apoptosis Caspase Pathway in Rat Ovarian Granulosa Cells in Vitro

Objective To investigate the effects of di-2-ethylhexyl phthalate (DEHP) on the expression of Caspase-3 and Caspase-9 genes, and on the apoptosis of rat ovarian granulosa cells in vitro.

Methods The ovarian granulosa cells of rats were separated; the primary cultured granulosa cells were treated with different concentrations of DEHP (0, 10, 50 and 100 nmol/L). The real time PCR was used to detect the mRNA of Caspase-3 and Caspase-9 genes, and the Western blot was used to detect the protein of Caspase-3 and Caspase-9 genes. Cell apoptosis rate was estimated by flow cytometry.

Results Compared with the control group, the expressions of Caspase-3 and Caspas-9 gene mRNAs and proteins in different DEHP-treated groups increased(P < 0.05) in a concentration dependent manner. The cell apoptosis rate in different DEHP-treated groups increased(P < 0.05) in a concentration dependent manner.

Conclusion DEHP could induce the apoptosis of ovarian granulosa cells through increasing the activity of caspase pathway, which results in ovarian dysfunction.