Bilotas M, Barañao R I, Buquet R, Sueldo C, Tesone M, Meresman G
Instituto de Biología y Medicina Experimental, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina.
Hum Reprod. 2007 Mar;22(3):644-53. doi: 10.1093/humrep/del423. Epub 2006 Nov 8.
Our purpose was to evaluate the effect of the GnRH agonist (GnRHa), leuprolide acetate (LA), and the GnRH antagonist (GnRHant), Antide, on apoptosis and expression of apoptosis-related proteins in endometrial epithelial cell (EEC) cultures from patients with endometriosis and controls (infertile women without endometriosis).
Biopsy specimens of eutopic endometrium were obtained from 22 patients with endometriosis and from 14 women that served as controls. Apoptosis was examined in EEC after incubation with LA and Antide. Bax, Bcl-2, Fas and FasL expression was evaluated after exposure to LA, Antide or a combination of both. The percentage of apoptotic cells (%ApC) was assessed by the acridine orange-ethidium bromide technique, and protein expression was evaluated by western blot and immunocytochemistry.
LA 100 and 1000 ng/ml increased the %ApC in EEC from patients with endometriosis (both P < 0.05) and controls (p < 0.05 and P < 0.01, respectively). Antide 10(-5) M increased the %ApC in EEC from patients with endometriosis and controls (P < 0.01). In EEC from women with endometriosis, Bax expression increased after treatment with LA, Antide and LA + Antide (P < 0.05, P < 0.001 and P < 0.001), whereas Bcl-2 expression decreased after exposure to LA and Antide (P < 0.001 and P < 0.01). FasL expression increased after LA, Antide and LA + Antide treatments (P < 0.01, P < 0.001 and P < 0.01). No significant changes were observed on Fas expression.
GnRH analogues enhanced apoptosis in EEC, and this was accompanied by an increase in expression of the pro-apoptotic proteins Bax and FasL and a decrease in expression of the anti-apoptotic protein Bcl-2.
我们的目的是评估促性腺激素释放激素激动剂（GnRHa）醋酸亮丙瑞林（LA）和促性腺激素释放激素拮抗剂（GnRHant）Antide对子宫内膜异位症患者及对照组（无子宫内膜异位症的不孕女性）子宫内膜上皮细胞（EEC）培养物中细胞凋亡及凋亡相关蛋白表达的影响。
从22例子宫内膜异位症患者和14例作为对照的女性中获取在位子宫内膜活检标本。将EEC与LA和Antide孵育后检测细胞凋亡情况。在暴露于LA、Antide或两者组合后评估Bax、Bcl-2、Fas和FasL的表达。通过吖啶橙-溴化乙锭技术评估凋亡细胞百分比（%ApC），并通过蛋白质印迹法和免疫细胞化学评估蛋白表达。
100和1000 ng/ml的LA增加了子宫内膜异位症患者（均P<0.05）和对照组（分别为P<0.05和P<0.01）EEC中的%ApC。10(-5) M的Antide增加了子宫内膜异位症患者和对照组EEC中的%ApC（P<0.01）。在子宫内膜异位症女性的EEC中，用LA、Antide和LA+Antide处理后Bax表达增加（P<0.05、P<0.001和P<0.001），而暴露于LA和Antide后Bcl-2表达降低（P<0.001和P<0.01）。LA、Antide和LA+Antide处理后FasL表达增加（P<0.01、P<0.001和P<0.01）。Fas表达未观察到显著变化。
GnRH类似物增强了EEC中的细胞凋亡，这伴随着促凋亡蛋白Bax和FasL表达的增加以及抗凋亡蛋白Bcl-2表达的降低。
