The Value Of AMH For Assessing Ovarian Function Injury Induced By Chemotherapy And Relevant Mechanism

Objective: This study was designed to explore the value of AMH for predicting ovarian reserve and assessing ovarian function injury induced by chemotherapy,and to clarify the protective effect of GnRHa on ovarian function exposed to chemotherapy with breast cancer as well as the mechanism of how GnRHa works through AMH.Materials,Patients and Methods:(1)Serum AMH levels were detected using ELISA assay in 177 healthy women and 30 normal female C57BL/6J mice of different ages,and the numbers of primordial follicles and growing follicles in mice of different ages were counted microscopically.(2)The clinical data were collected from 72 premenopausal women with breast cancer that conform to chemotherapy,the serum AMH levels were tested pre-and post-chemo whereafter the data was analysized to evaluate ovarian function injury.(3)Seventy-five Balb/c nude mice with breast cancer were randomly assigned to five groups: Control,CTX 100mg/kg,GnRHa + CTX 100mg/kg,CTX 200mg/kg,GnRHa + CTX 200mg/kg.Serum AMH levels were detected before chemotherapy and 1,3 and 5 weeks after treatment,respectively.The ovaries from mice at different stages were collected,whereafter the numbers of primitive follicles and growing follicles were recorded.(4)In addition,human ovarian granulosa cells were treated with CTX and GnRHa,and protein immunoblotting,ELIS and other techniques were used to observe AMH secretion changes at protein level.Results:(1)In this study,we found that the decline rate of AMH was accelerated in mice older than 8 months and women over 35 years old,and the reduction of the primordial follicles and growing follicles in mice was directly related to the AMH level.(2)The AMH level,as well as the numbers of primordial follicles and growing follicles showed a significant decrease in mice treated with high-dose CTX,while the changes were not significant in the low-dose CTX group.In breast cancer patients treated with chemotherapy alone,there was no significant difference on serum AMH level between pre-and post-chemo in patients younger than 35 years old,while the serum AMH level decreased significantly in patients over 35 years old.(3)After the combination of CTX and GnRHa,the serum AMH of breast cancer mice remained stable after a slight decrease.The discrepancy of AMH between mice treated with and without GnRHa was great,especially in the high-dose CTX group.The numbers of primordial follicles and growing follicles were higher in the combination group than that in the CTX alone group.(4)In granulosa cells treated with CTX,the expression of intracellular AMH was increased,while the secretion of AMH in the culture medium decreased significantly.Magically,this phenomenon could be reversed by GnRHa treatment.Conclusion: Our study indicates that serum AMH is an excellent biomarker of ovarian reserves,which predicting ovarian aging and ovarian injury precisely in mature mice and women.The predicting value of AMH may be more effective in premenopausal women accepted chemotherapy over 35 years old.It was also found that GnRHa could reverse the inhibition of AMH secretory induced by CTX,and then increase the serum AMH level so as to prevent the development of primordial follicles into growing follicles,which resulted in a stable ovarian reserves.Thus we conclude that AMH plays an important role in ovarian protection conducted by GnRHa.