| The objective is to investigate the effect of Genistein on ovarian reserve in young and initial aging Sprague-Dawley(SD)female rats of different age groups.Young female rats(200±20g,n=40),aged 49 days and initial aging female rats(490±20g,n=40),aged 10-11 months were selected and according to weight,they were divided into the following five groups with 8 animals in each: negative control group(NC),low dose group(L),middle dose group(M),high dose group(H)and positive control group(PC).GEN group received GEN of 15,30,60 mg/kg respectively,PC group received diethylstilbestrol of 0.5 mg/kg.It lasted 30 days.After the experiment,the body weight and the main organs weight of rats were measured.Serum levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH)and inhibin A(INH-A),inhibin B(INH-B),anti-mullerian hormone(AMH),insulin-like growth factors-1(IGF-1)and insulin-like growth factor binding protein-1(IGFBP-1))were detected by enzyme linked immunosorbent assay(ELISA)method.The mRNA expression levels of insulin-like growth factors(IGF),steroidogenic acute regulatory protein(StAR),P450 side-chain cleavage enzyme(P450scc),aromatase cytochrome P450(P450arom)、CYP19,bone morphogenetic protein-15(BMP15)and Growth differentiation factor-9(GDF-9),early growth response gene-1(EGR-1)and stromal cell-derived factor(SDF-1)in ovary tissue were detected by Realtime fluorescence quantitative PCR(RTFQ PCR).The protein expression levels of StAR、P450scc、P450arom and EGR-1 in ovary tissue were detected by Western Blot(WB).The results are as follows:(1)GEN had no significant effect on body weight,liver/body ratio and kidney/body ratio of young and initial aging rats(P>0.05).But,the spleen/body ratio increased,the uterine/body ratio and ovarian/body ratio decreased(P<0.05 or P<0.01)in M and H groups.(2)The effect of GEN on gonadotropin FSH and LH in the serum of young and initial aging female rats was opposite.With the increase of GEN does,the serum contents of FSH decreased(P<0.05);the serum contents of LH decreased too,but the difference was not significant(P>0.05).(3)The effect of GEN on cytokines: The result of serum inhibin levels showed that INH-B decreased in young group rats(P<0.01).AMH decreased in both young and initial aging rats,but levels in the young rats were more obvious(P<0.05 or P<0.01);IGFBP-1 levels increased significantly in serum(P<0.05 or P<0.01),the expression of IGF-1 and IGFBP-1 mRNA in the ovary increased in M and H dose of GEN(P<0.01);the increase of BMP-15 in the serum of the young group was significant(P < 0.05 or P < 0.01),there was no significant change of BMP-15 mRNA in the ovary(P>0.05);The expression of GDF-9 in the serum and the ovaries of young and initial aging rats increased(P<0.05 or P<0.01).(4)The effect of GEN on the key enzyme gene of estrogen production: GEN can significantly improve StAR,P450 scc,P450arom,CYP19 gene and protein expression(P<0.05 or P<0.01).The expression of estrogen effector gene EGR-1 increased significantly(P<0.05 or P<0.01).The expression of SDF-1 mRNA increased in M and H groups of young and initial aging rats(P<0.05 or P<0.01).Conclusion: GEN could regulate the expression of key enzyme genes of estrogen by regulating androgengenesis.It could affect ovarian hormone production,regulate the expression of follicular development-related factors.So the growth and development of ovarian follicle could be regulated.The same dose of GEN had a slight difference on young and initial aging female rats.GEN had some effect on the ovaries of young females,but follicle viability reduce.For initial aging female rats,GEN had a positive effect on ovarian reserve function and could improve the follicle viability. |
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