| objectiveIn this experiment we observed the effect of mefipristone on apoptosis in cultured granulosa cell's with mefipristone- treatment, in order to investigate the relationship between mifepristone and follicular development and possible mechanisms which are treated sexual hormone-dependent disease by Mifepristone.MethodsFour-five week female rats were injected peritoneal with 60IU PMSG to irritate follicle growth, after 48 hours rats were sacrificed and GCs were separated from follicular fluids. Electron microscopy was used to identify GC, flow cytometry and fluorometry were conducted to observe the biological and morphologic changes of granulosa cells exposed to different concentrations of mifepristone for 24 hours (0,10-6,10-5 and 10-4mol/L mifepristone), Immunocytochemistry was performed to detect the Fas expressions separately, the estradiol and progestrone lever of serum were detected by chemiluminescence.One-way ANOVA and least significant difference test were used for statistical analysis. Two-tailed test with P＜0.05 was considered significant. All data were managed with SPSS 11.5 for windows.Results1. Rat granulosa cells were successfully obtaind for primary cell culture, 90% of which are lively. The cells were well spread within the 24 h, showing numerous stress fibers and numerous cytoplasmic lipid droplets characteristic of steroidogenic cells. Identification of GC: Distinctive ultramicrostructure of GC can be found by transmission electron microscope: tubiform chondrosome, SER and adipoid is abundant, and the quantity of GCs reach 80%-85%.2. The apoptosis rate of Granulosa cells in mifepristone-treated cells were increased, low dose groups (6.7%±0.73%) had no significant difference compared with that of control group (7.5%±0.88%) (P＞0.05), moderate (9.53%±0.78%) and high (13%±1.75%) treated groups had the significant difference compared with that of control group(P＜0.05). The quantity of GC with chromatoid cell nuclus were increased with increased concentration of mifepristone in fluorescence microscope, low dose groups (4.18%±0.90%) had no significant difference compared with that of control group (1.49±2.20%) (P＞0.05), moderateand (6.63%±1.48%) and high (15.45%±3.89%) treated groups had the significant difference compared with that of control group(P＜0.05).3. The expression of Fas in mifepristone-treated cells showed significant increase, compared with that of control group (P＜0.01), and comparisons among the different treated groups had the significant difference.4. The descendent Progestrone level has dose-dependant relationship with concentration of mifepristone, compared with that of control group(P＜0.05), and comparisons among the different treated groups had the significant difference. Mifepristone had no direct effect on estradiol, and there is no significant difference between groups.Conclusions1. Mifepristone was able to induce apoptosis in cultured granulose cells, The apoptosis rate of GC may be dose-dependant with Mifepristone, which maybe the mechanisms Mifepristone treating sexual hormone-dependent disease.2. Mifepristone reduce progesterone, production maybe by Regulating the expression of Fas protein, which maybe is one of the mechanisms in inducing apoptosis of granulosa cells and inhibiting ovulation.3. Mifepristone had no evident inhibition on estradiol lever in cultured granulose cells, which maybe indicate that the secreted function of granulose cells couldn't be significant affected by mifepristone.
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