Role of apoptosis on human follicular growth and analyzes of regulattry mechanism of the apoptosis in granulosa cells

1)我们用TUNEL法分析了体外受精时获得的人颗粒细胞的凋亡。培养颗粒细胞，分析培养液中加入或不加入hCG时颗粒细胞凋亡发生的时间过程，但在整个过程中，部分样品显示凋亡，而另一些样品则未显示凋亡。未观察到hCG的影响。这些结果表明，颗粒细胞或黄体细胞的凋亡可能不参与这些细胞的细胞转换调节。2）为了阐明凋亡是否参与子宫内膜异位症，我们从12例子宫腺肌病和12例卵巢中获得了在位子宫内膜组织沿着增生组织。用TUNEL法检测凋亡诱导的DNA片段化，并用抗Fas、Le^y或bcl-2的单克隆抗体对同一组织切片进行免疫染色。分析表明，凋亡参与了所有的卵巢增生组织，但只有2/12的腺肌病组织和5/24的在位子宫内膜组织。这些病例均与月经周期的阶段无关。bcl-2在在位子宫内膜和腺肌组织中的表达仅限于增殖期，在12例卵巢子宫内膜异位症中仅1例有表达。Fas和Le^y在在位和异位子宫内膜组织中广泛表达。这些结果表明，卵巢子宫内膜异位症的特点是不同的子宫腺肌病和在位子宫内膜的细胞凋亡参与。

1) We analyzed apotosis by TUNEL method in the human granulosa cells obtained when in vitro fertilization was performed. The granulosa cells were cultured, and time-couse of apoptosis occurrence were analyzed with or without hCG in the culture media, however, some samples showed apoptosis and the others showed no apoptosis throughout the course. No effects of hCG were observed. These results indicated that the apoptosis in the granulosa or luteal cells might not involved the regulation of cell turn-over in these cells.2) To clarify whether apoptosis is involved endometriosis, we obtained eutopic endometrial tissues along with endometriotic tissues from the adenomyosis in 12 cases and from the ovary in 12 other cases. Apoptosis-induced DNA fragmentation was detected by the TUNEL method, and immunostaining with a monoclonal antibody against the Fas, Le^y or bcl-2 was also performed using the same tissue section. Analysis showed that apoptosis was involved in all the ovarian endometriotic tissues but in only 2 of the 12 adenomyotic tissues and in 5 of the 24 eutopic endometrial tissues. None of these cases correlated to phases of the menstrual cycle. The expression of bcl-2 in the eutopic endometrial and adenomyotic tissues was limited to be proliferative phase, and was observed in only one of the 12 cases of ovarian endometriosis. Fas and Le^y were expressed randomly across a wide range in both the eutopic and ectopic endometrial tissues. These results suggest that features of ovarian endometriosis is defferent from adenomyosis and eutopic endometrium in terms of apoptosis involvement.