Effect Of Gnistein On FSHR And AMH In Ovaries Of Subacutely Aging Mice

By establishing a subacute aging animal model,we explored the effects of genistein(GEN)on the anti-mullerian hormone(AMH)and follicle stimulating hormone receptor(FSHR).Seventy-two 2.5-month-old female Kunming mice(30 ± 2g)were selected and randomly divided into 6 groups according to body weight: blank control group(CG),model group(MG),GEN low(L),medium(M),High-dose group(H)and estradiol valerate control group(EG).In addition to the blank group,the other groups were injected intraperitoneally with 200 mg / kg of D-galactose physiological saline to prepare a subacute aging mouse model.After successful modeling,the GEN low,middle,and high dose groups were given 10,20,and 40 mg / kg of GEN perfusion.In the stomach,the positive control group was administrated with 0.5 mg / kg of estradiol valerate,and the blank group was administrated with the same amount of peanut oil for a period of 15 days.The mice were weighed during the experiment,and important organs were weighed after dissection.The estrous cycle and ovarian morphology of the mice were observed and the follicle counted.The enzymes linked immunosorbent assay(ELISA)was used to detect serum AMH,Estradiol(E2),Follicle stimulating hormone(FSH),Luteinizing hormone(LH),Progesterone(P),Stromal cell-derived factor,SDF-1 / CXCL12).Detect the gene level of AMH,Anti-mullerian hormone receptor typeⅡ(AMHRⅡ),FSHR,LH,LHCGR,SDF-1,Chemokine receptor 4(CXCR4)in mouse ovarian tissue by real-time fluorescence quantitative PCR(RTFQ-PCR)technology.Western blot(WB)was used to detect the protein levels of AMH and AMHRⅡ,FSH and FSHR,LH and LHCGR,CXCL12 and CXCR4 in mouse ovaries.Experimental results:(1)The ovaries in the GEN dose group were larger than the model group,and the ovarian length of the GEN high dose group(40 mg / kg)was longer than that in the model group,with a significant difference(p <0.05);(2)The number of sinusoidal follicles in each dose group of GEN is lower than that in the model group,and the difference is extremely significant((p <0.01);the estrous period of model mice is disordered,and the estrous period is longer than that of the normal group.The difference is significant(p<0.05);(3)After the end of modeling,the weight of the model group decreased compared with the blank group,and the difference was significant or extremely significant(p <0.05,p <0.01).After the start of gastric administration,the body weight of the model group was not significantly different from that of the GEN dose groups.;There was no significant difference in ovaries / weight and uterus / weight between the GEN dose groups and the model group;(4)The serum AMH concentration of model mice was lower than that of the blank group,and the difference was significant or extremely significant(p <0.05,p <0.01).The GEN low and high dose groups were higher than the model group after gavage,and the differences were significant(p <0.05);Serum FSH and LH concentrations of model mice were lower than those of the blank group,and the differences were significant or extremely significant(p <0.05,p <0.01).FSH and LH concentrations of GEN dose groups were lower than those of the model group after gavage,the difference was significant or extremely significant(p <0.05,p <0.01);the concentrations of serum E2,P,and CXCL12 in model mice were lower than those in the blank group,and the difference was significant or extremely significant(p <0.05,p <0.01).The post-gastric GEN dose groups were higher than the model group,and the differences were significant or extremely significant(p <0.05,p <0.01);(5)The expression levels of AMH and LH m RNA in the middle dose of GEN were higher than those in the model group,with a significant difference(p <0.05).The expression levels of AMHRⅡ and LHCGR m RNA in the middle dose group of GEN were higher than those of other dose groups,but lower In the model group;as the concentration of GEN increased,the expression of FSHR m RNA also showed an upward trend,in which the expression level of the GEN high-dose group was higher than that of the model group;the expression of CXCL12 m RNA was highest at the high-gen GEN level,and it The difference was significant(p <0.05).The expression of CXCR4 m RNA gradually increased with the increase of GEN dose,and the expression was highest at high doses of GEN;(6)The expression levels of AMH,AMHRⅡ and LH,LHCGR proteins in mouse ovaries increased with the increase of GEN dose,which was significantly different from the model group(p <0.01).The expression level of FSH,FSHR protein and CXCL12,CXCR4 protein also increased with the increase of GEN concentration,which was significantly different from the model group(p <0.01),of which GEN high dose The expression level was also highest,and the protein expression level was higher than that of the estrogen control group.Conclusion: This shows that GEN has a good regulatory effect on the levels of endocrine-related hormones in the serum,and can improve the m RNA and protein expression levels of ovarian-related factors AMH and FSHR.GEN regulates the levels of ovarian-related factors such as AMH and FSHR,and then plays a role in regulating ovarian growth and development,and ultimately achieves the purpose of improving ovarian function.