GnRHa对子宫内膜异位症内膜间质细胞生长增殖及血管形成的影响

Effect of GnRHa on the proliferation and angiogenesis of ectopic and eutopic stromal cells

目的:研究GnRHa对子宫内膜异位症异位及在位内膜间质细胞增殖、凋亡及血管形成的影响。方法:体外原代培养人内异症异位及在位子宫内膜间质细胞,用不同浓度的GnRHa分别干预,四甲基偶氮噻唑蓝(MTT)法测定内膜间质细胞的生长增殖活力;Annexin V-FITC和PI双染、流式细胞仪(FCM)检测异位及在位内膜间质细胞凋亡及周期变化;免疫细胞化学检测血管内皮生长因子(VEGF)表达的改变。结果:GnRHa可抑制内异症异位及在位内膜间质细胞增殖且呈剂量依赖关系;GnRHa能明显增加异位及在位内膜细胞的凋亡率(P<0.05),且异位内膜细胞凋亡率明显高于在位内膜(P<0.05);不同浓度的GnRHa作用后,内膜间质细胞均发生细胞周期阻滞,G1期细胞增加,S期细胞减少,此作用随浓度增加而增强(P<0.05);免疫细胞化学测定表明,GnRHa可降低异位及在位内膜间质细胞VEGF的表达(P<0.05),且随GnRHa浓度增加而增强(P<0.05)。结论:GnRHa可明显抑制人内异症异位及在位子宫内膜间质细胞的生长并促进其凋亡,降低异位及在位内膜间质细胞VEGF的表达。

Objective：To investigate the effect of GnRHa on the proliferation and angiogenesis of ectopic and eutopic endometrial stromal cells from women with endometriosis in vitro. Methods：The ectopic and eutopic endometrial stromal cells were isolated and cultured in vitro, then exposed to different concentration of GnRHa. The growth and proliferation of ectopic and eutopic stromal cells were measured by MTT assay. The cell cycle distribution and apoptosis of ectopic and eutopic endometrial stromal cells were measured by flow cytometry with Annexin V-FITC and PI double staining. The expression of VEGF induced by GnRHa was analyzed by immunocytochemistry. Results：GnRHa could inhibit the proliferation of ectopic and eutopic endometrial stromal cells from women with endometriosis in a dose dependent manner. GnRHa increased the apoptotic rate of both ectopic and eutopic cells, and blocked the cell cycle in a dose dependent manner（ P 〈 0.05 ）. The apoptotic rate of ectopic cells was absolutely higher than that of eutopic cells. The number of ectopic and eutopic cells were increased in G1 stage and decreased in S stage. The expression of VEGF in ectopic and eutopic cells were significantly decreased after the treatment of GnRHa（P 〈 0.05 ）, this action was more significant in the highconcentration group than in the low-concentration group （ P 〈 0.05 ）. Conclusion： GnRHa can significantly inhibit the proliferation and enhance the apoptosis of ectopic and eutopic endometrial stromal cells from women with endometriosis, and decrease the expression of VEGF.