Résumé
Objective To explore the effects of low-dose mifepristone on human granulosa cells and the molecular mechanism of its inhibiting or delaying ovulation.Methods Phase contrast microscopy and electron microscopy were conducted to observe the morphological and biochemical change of granulosa cells exposed to various doses of mifepristone for 24 hours(the final concentrations of mifepristone were respectively 2.5?mol/L,5?mol/L and 10?mol/L,Non-mifepristone as the control group).Immunocytochemistry was performed to detect the protein expressions of Bcl-2 and Bax in granulosa cells exposed to various doses of mifepristone for 24 hours.The mRNA levels of Bcl-2 and Bax in granulosa cells were detected by RT-PCR.Results Granulosa cells treated with various doses of mifepristone presented typical morphological characteristics of apoptosis such as condensation of chromatin and cytosolic,and shrink of cells.The expression of Bcl-2 protein might be down-regulated,while the expression of Bax might be up-regulated by mifepristone.The histological score(HSCORE) of Bcl-2 protein in granulosa cells,which were incubated with various doses of mifepristone(2.5?mol/L,5?mol/L,10?mol/L) for 24 hours,were respectively 2.15?0.16,1.88?0.13 and 1.64?0.16 respectively,Compared with the HSCORE of the control group(2.51?0.16),the difference was significant(P0.05).Conclusion Mifepristone could induce human granulosa cells apoptosis.The Bcl-2 protein and Bax protein may be involved in the apoptosis of granulosa cells induced by mifepristone.
Résumé
AIM:To investigate the effect of different low-dose mifepristone on apoptosis in granulosa cells and to test low-dose mifepristone as an orally contraceptive drug.METHODS:By using immunofluorescence,terminal deoxynucleotidyl transferase-mediated nick end labelling(TUNEL)and flow cytometry technique,the nuclear morphologic features and ratio of apoptosis and fluorescent intensity of caspase-3 in granulosa cells cultured in vitro treated with different low-doses of mifepristone were observed,respectively.RESULTS:By the display of immunofluorescence,the granulosa cells in treatment group were classified as apoptotic cells on the basis of their morphologic features contained a single condensed chromatin,multiple nuclear fragments.The results of TUNEL showed significant difference between control group and groups treated with different concentration of mifepristone(P