Effect of GnRHa on apoptosis and release of VEGF in endometrial cell cultures from patients with adenomyosis
WANG Xiao-xia1,KANG Jia-li1,SHAO Xue-fei2,YU Jia1,DENG Ling-hong1 Department of Gynecology and Obstetrics,The First Municipal People' s Hospital Affiliated to Guangzhou Medical College,Guangzhou 510180,China
AIM:To investigate the effect of Gonadotropin-realeasing hormone agonist(GnRHa) on apoptosis and the release of vascular endothelial growth factor(VEGF) in the in vitro eutopic endometrial cell of adenomyosis.METHODS:Biopsy specimens of eutopic endometrium obtained from 32 women with adenomyosis and 20 normal women were studied.Cells were cultured with GnRHa for 24 h,48 h and 72 h in the concentration of 10-7 mol/L and 10-5 mol/L.Apoptotic ratio was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) and flow cytometry before and after using GnRHa.VEGF concentrations in culture supernatant were detected by commercial enzyme-linked immunosorbent assay(ELISA).RESULTS:(1)Cells in two groups showed cell shrinkage,floating,nucleus concentration,nuClear fragmentation and typical apoptotic bodies.(2)The apoptotic ratio of cultured AM endometrial cell without GnRHa was lower than that in control group and the apoptotic ratio increased with the prolongation of time in two groups(P0.01).(3)After GnRHa addition,each group showed higher apoptotic ratio and a trend towards higher apoptotic ratio linked to advanced concentration.Furthermore,apoptotic ratio in study group was significantly higher than that in control group at the same time point and same concentration(P0.01).(4) The concentrations of VEGF in the eutopic endometrial cells of adenomyosis were significantly higher than those in the normal endometrimn.The concentrations of VEGF in the both groups were downregulated by GnRHa in a dose-dependent manner.CONCLUSION:(1) The abnormality of apoptotic ratio of AM endometrial cells may be associated with the AM pathogenesis.GnRHa nmy increase the apoptotic ratio of cultured AM endometrial cells by autocrine or paracrine.(2)VEGF may play an impotant role in the pathogenesis of adenomyosim.GnRHa can directly suppress the survival and growth of ectopic endometrial by decreasing the release of VEGF which was related to the adenomyosis angiogenesis.