Effect Of Bone Morphogenetic Protein-9 On The Expression Of Follicle Stimulating Hormone Receptor And Luteinizing Hormone Receptor In Human Ovarian Ranulose Cells

Objective: To investigate the effect of bone morphogenetic protein-9 on the expression of follicle stimulating hormone receptor and luteinizing hormone receptor in human ovarian luteinized granulosa cells and to explore the transduction mode of Smads in transforming growth factor-? signal pathway.Methods: Follicular fluid was collected from patients undergoing in vitro fertilization or intracytoplasmic sperm injection-embryo transfer due to fallopian tubes factors or male sperm factor,the uteinized granulosa cells were extracted and cultured and were identified by immunocytochemistry.The m RNA expression levels of FSHR and LHR in granulosa cells reated with BMP-9 were detected by real-time quantitative polymerase chain reaction,Western-blot method was used to detect the effect of BMP-9 on the protein expression of FSHR,LHR and Smads in granulosa cells.Results: FSHR was localized in the cytoplasm of granulosa cells,and the positive staining rate was more than 95%.The results of q RT-PCR showed that compared with the control group,the expression of FSHR m RNA in granulosa cells was significantly up-regulated in BMP-9 group,and the difference was statistically significant(P<0.001),the expression of LHR m RNA was inhibited after BMP-9 treatment,and the difference was statistically significant(P<0.05).The results of Western-blot showed that compared with the control group,BMP-9 significantly increased the expression of FSHR protein in granulosa cells,and the difference was statistically significant(0.09 ±0.01,0.19 ±0.03,P<0.05),the expression of LHR protein was inhibited,but there was no significant difference(0.58 ±0.03,0.52 ±0.04,P>0.05),BMP-9 significantly increased the expression of p-Smad1/5/9 protein(0.33 ±0.01,0.84 ±0.08,P < 0.05),BMP-9 had no effect on the expression of p-Smad2/3,and the difference was not statistically significant(0.34 ±0.04,0.29 ±0.06,P>0.05).It is suggested that BMP-9 is easy to activate Smad1/5/9 signal transduction pathway and play a signal transduction role in granulosa cells.Conclusion: This study found that the effect of BMP-9 on the expression of FSHR and LHR and its role in the conduction pathway may provide new ideas for regulating follicular development and oocyte maturation.