GnRHa对子宫腺肌病在位内膜细胞凋亡及VEGF分泌的影响

目的:探讨促性腺激素释放激动剂(GnRHa)对子宫腺肌病(AM)在位内膜细胞凋亡及血管内皮生长因子(VEGF)分泌的影响。方法:取32例AM患者在位内膜细胞及20例对照组正常子宫内膜细胞进行体外培养,并予不同浓度GnRHa处理细胞,分别作用24、48、72 h后,用末端转移酶介导的缺口末端标记法(TUNEL)及流式细胞仪检测各组细胞的凋亡率;采用ELISA法检测细胞培养上清液中VEGF的含量。结果:(1)镜下可见两组部分细胞固缩、漂浮、核浓缩、碎裂,呈典型的凋亡小体;(2)不含GnRHa培养的AM在位内膜细胞凋亡率在各时间点均低于对照组,且两组都随时间的延长凋亡率增加(P<0.01);(3)GnRHa作用后,两组的凋亡率高于作用前,且均随浓度的增加而增加,且实验组各时间点、各浓度AM在位内膜细胞凋亡率均显著高于同时间、同浓度的对照组(P<0.01);(4)VEGF在AM在位内膜细胞中的分泌高于正常子宫内膜细胞(P<0.05),且GnRHa呈剂量依赖方式抑制AM在位内膜细胞及正常子宫内膜细胞VEGF的分泌。结论:(1)在位内膜细胞凋亡率异常可能是AM发病机制之一。GnRHa可能以自分泌或旁分泌机制直接作用于在位内膜细胞,提高了AM在位内膜细胞的凋亡率。(2)VEGF可能与子宫腺肌病的发生发展有关。GnRHa对子宫内膜细胞有直接作用,可能通过抑制VEGF的分泌,阻碍血管生成,从而达到抑制子宫内膜在子宫肌层内异位种植和生长的目的。

Effect of GnRHa on apoptosis and release of VEGF in endometrial cell cultures from patients with adenomyosis

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(P<0.01).(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(P<0.01).(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.