| Objective: In the process of in vitro fertilization-embryo transfer(IVF-ET),physiological processes such as development,maturation and ovulation of dominant follicles are important.BMP-9 is a member of BMP,and the BMP family is known to act broadly on ovarian germ cells,showing dynamic changes in follicular development and corpus luteum formation.Recent studies have shown that BMP-9 is expressed in human ovarian granulosa cells.We will investigate the effect of BMP-9 on estrogen(E2)and progesterone(P)secretion of human luteinized granulose cell in vitro.Methods: Subjects Follicular fluid were collected from the women undergoing IVF-ET in the forth hospital of HeBei medical university Reproductive Medicine between April 2017 to May 2017.Human luteinized granulose cells from follicular fluid were purified and then identificated by immunohistochemistry.Then they were cultured with medium plus variousc oncentration of BMP-9(0,3,6,12 ng/ml)and FSH 75IU/L or alone for 72 h.The media were collected to measure the concentration of E2 and P by ELISA.And western-blot method was used to determine the relative expression of P450 arom and StAR proteins.Statistical analysis:All data were analyzed by SPSS21.0 statistical software.Data were presented as (?)±s.The comparisons between various groups were performed by t test.A P value of 0.05 was considered as statistically significant.Result:1.More than 90% of the luteinized granulosa cells were positively stained for FSH receptors.2.Luteinized granulosa cells could autocrine E2 and P without BMP-9 or FSH co-culture.3.When different concentrations of BMP-9 were cultured alone,there was no significant difference in E2 and P concentrations and basal levels(0ng/ml BMP-9);4.Different concentrations of BMP-9 co-cultured with FSH had no significant effect on the secretion of E2,but could inhibit the secretion of P.5.When cultured with different concentrations of BMP-9,the expression levels of P450 arom protein and StAR protein were not statistically significant compared with the basic state;6.Different concentrations of BMP-9 co-cultured with FSH had no significant effect on the expression of P450 aromatase protein,but it could down-regulate the expression of StAR protein.Conclusion:Addition of different concentrations of BMP-9 had no significant effect on E2 and P production by luteinized granulose cells in the absence of FSH;Different concentrations of BMP-9 combined with FSH caused a dosedependent inhibition of P production by luteinized granulose cells,but had no effect on E2 production;BMP-9 can inhibit the expression of StAR protein without FSH stimulation without affecting the expression of P450 arom protein;BMP-9 may act as a luteinizing inhibitor to prevent the follicles in advance of luteinization,and regulate follicular development. |
