Correlation Factors And Mechanisms Involved In The Developmental Anomaly Of Follicles In Polycystic Ovary Syndrome

PartⅠ.The correlation between follicle count and clinical characteristics and metabolic endocrine anomalies in polycystic ovary syndrome Purpose:Investigating the correlatin between follicle count and clinical characteristics and endocrine metabolic parameters to explore the reason for follicular excess in PCOSMethods:Sixty-four cases with PCOS were included.Cases were evaluated for their clinical history,body weight,body height,FSH,LH,testosterone,DHEAS, total cholesterol,triglyceride,HDL,LDL,fasting plasma glucose,fasting insulin,AMH,follicular count and ovarian volume,FAI,and HOMA.SPSS 15.0 was used for Pearson correlation and regression analysis.A P vulue less than 0.05 is regarded as significant.Results:Follicle count is positively correlated with FAI.(correlation coefficient:0.264,P=0.038,regression equation:y+0.638x+17.237),negatively correlated with SHBG(correlation coefficient:-0.317,P=0.019,regression equation:y=-0.026x+23.611),positively correlated with AMH(correlation coefficient:0.322,P=0.011,regression equation:y=0.207+17.468),and positively correlated with ovarian volume(correlation coefficient:0.322, P=0.011,regression equation:y=0.207x+17.468).The correlation between follicle count and other parameters(age,BMI,WHR,FSH,LH,DHEAS,TC,TG,HDL, LDL,fasting plasma glucose,and fasting insulin) are not significant different. Conclusion:Androgen,ovarian volume,and AMH are significantly correlated with follicle count.Among them,the increase of ovarian volume and serum AMH in PCOS may be the results of follicular excess,and hyperandrogen may contribute to the increase of follicle count thus deserve further investigation. PartⅡ.The effects of testosterone,insulin,and luteinizing hormone on the expression of anti-M(u|¨)llerian hormone in in-vitro cultured rat granulosa cells of preantral folliclesPurpose:Investigating the effects of androgen,insulin,and LH on the AMH expression in granulose cells to explore the mechanism of follicular excess in PCOSMethods:Granulosa cells from rat preantral follicle were cultured in-vitro. 1.10^-7M testosterone,100ng/mL insulin,or 10ng/mL LH were added to the culture medium.AMH mRNA expression was determined 24 hours later.2.Time-course analysis,10^-7M testosterone was added to the culture medium.The expression of AMH mRNA and AMH typeⅡreceptor mRNA were determined 0,1,4,12,and 24 hours later.3.Dose-response analysis.10^-9M to 10^-5M testosterone were added to the culture medium respectively.AMH mRNA expression was determined 12 hours later. 4.10^-7M testosterone was added to the culture medium.Cell-conditioned medium were obtained 0,1,2,and 3 days later to determine the AMH concentration by ELISA.Result:1.Testosterone significantly inhibited the expression of AMH mRNA (P＜0.05).Insulin and LH did not affect the expression of AMH mRNA.2.The longer the stimulation of testosterone,the lower expression level of AMH mRNA and AMN typeⅡreceptor mRNA,with the nadir at 12 hours.3.The higher the concentration of testosterone,the lower the expression level of AMH mRNA.4.The secretion of AMH in culture medium significantly decreased after 3 days of testosterone stimulation.Conclusion:Testosterone inhibited the expression of AMH and AMH typeⅡreceptor mRNA in in-vitro cultured granulose cells from rat preantral follicles. The follicular excess in PCOS may be resulted through the same mechanism. Hyperandrogen in PCOS may suppress the AMH function in ovary,thus lead to the overgrowth of preantral and antral follicle,and plycystic ovary may ensue.PartⅢ.The effects of anti-M(u|¨)llerian hormone on the expression of follicular stimulating hormone receptor and aromatase in in-vitro cultured granulose cells of rat antral folliclesPurpose:Investigating the effect of AMH on the sensitivity of granulosa cells to FSH to explore the reason for decreased response of granulose cells to FSH in PCOSMethod:Granulosa cells from rat antral follicle were cultured in-vitro,1. Cells were pre-treated with FSH 10ng/mL for 24 hours,and were supplemented with LH 10ng/mL or AMH 1000ng/mh thereafter.Aromatase mRNA expression was determined 24 hours later.2.Cells were pre-treated with FSH 10ng/mL for 24 hours,and were supplemented with AMH(10,100,or 1000ng/mh,respectively).Aormatase mRNA and protein expression level were determined 24 hours later.Result:LH significantly increases the expression of aromatase mRNA,and AMH suppressed the expression of aromatase mRNA.2.The higher the concentration of AMH,the lower the expression of aromatase mRNA and protein.The adding of AMH did not significantly affect the expression of FSH receptor mRNA.Conclusion:The decrease of sensitivity to FSH by AMH in granulose cells is not resulted through the suppression of FSH receptor.AMH decreases sensitivity to FSH only at a higher concentration,which implied that the failure of formation of dominant follicle will come from the local effect of AMH but not the higher serum AMH.