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首页> 外文期刊>Journal of the Society for Gynecologic Investigation >Expression of mitochondria-dependent apoptosis genes (p53, Bax, and Bcl-2) in rat granulosa cells during follicular development.
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Expression of mitochondria-dependent apoptosis genes (p53, Bax, and Bcl-2) in rat granulosa cells during follicular development.

机译：卵泡发育过程中大鼠颗粒细胞中线粒体依赖性凋亡基因（p53，Bax和Bcl-2）的表达。

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OBJECTIVE: We examined rat ovarian granulosa cells at different follicular stages and evaluated the apoptosis pattern of the mitochondria-dependent genes during folliculogenesis. METHODS: After down-regulating ovarian function with gonadotropin-releasing hormone agonist (GnRHa), granulosa cells were collected from the rat ovary at different stages of the following different hormonal treatment paradigms: stage E (after estrogen treatment), EF (after E + follicle-stimulating hormone [FSH] treatment), and EF hCG (after E + FSH + human chorionic gonadotropin treatment). To evaluate the in vitro susceptibility of granulosa cells at different developmental stages to apoptosis, the collected cells were cultured in a serum-free medium with or without E2 for 24 hours. The regulation of apoptosis in the granulosa cells was analyzed using fluorescein-activated cell sorting, quantitative competitive polymerase chain reaction, and western blot methods. RESULTS: The apoptosis rate of the freshly isolated granulosa cells tended to increase according to the hormonal treatment paradigm. In addition, during the hormone treatment, mitochondria-dependent apoptosis genes showed the following changes: although the Bax mRNA level did not change, the Bcl-2 mRNA level decreased significantly (P <.05). The p53 mRNA level increased significantly (P <.05) and closely matched the apoptosis rate (R = 0.7, P <.05). The expression of the active form of the caspase-3 protein (the final executioner of cell death) tended to increase and showed a good correlation with the apoptosis rate (R = 0.96, P <.01). After an in vitro culture of the granulosa cells, the apoptosis rate tended to increase at all stages, particularly stage EF hCG (P <.05). Bax and p53 mRNA tended to increase and showed a good correlation with the apoptosis rate (R = 0.64, P <.05 and 0.86, P <.01). The Bcl-2 mRNA level tended to decrease at all stages showing no correlation with the apoptosis rate. The expression level of the active caspase-3 protein tended to increase atall stages and showed a good correlation with the apoptosis rate (R = 0.93, P <.01). CONCLUSION: Apoptosis of rat ovarian granulosa cells tends to increase according to the stage of follicular development. Among the mitochondria-dependent genes, p53 closely correlates with granulosa cell apoptosis during follicular development.
机译：目的：我们研究了不同卵泡阶段大鼠卵巢颗粒细胞，并评估了卵泡形成过程中线粒体依赖性基因的凋亡模式。方法：用促性腺激素释放激素激动剂（GnRHa）下调卵巢功能后，在以下不同荷尔蒙治疗范例的不同阶段从大鼠卵巢收集颗粒细胞：E期（雌激素治疗后），EF（E +之后）促卵泡激素[FSH]治疗）和EF hCG（E + FSH +人绒毛膜促性腺激素治疗后）。为了评估不同发育阶段的颗粒细胞对凋亡的体外敏感性，将收集的细胞在有或没有E2的无血清培养基中培养24小时。使用荧光素激活的细胞分选，定量竞争聚合酶链反应和蛋白质印迹方法分析了颗粒细胞的凋亡调控。结果：根据荷尔蒙治疗模式，新鲜分离的颗粒细胞的凋亡率趋于增加。此外，在激素治疗期间，线粒体依赖性凋亡基因显示以下变化：尽管Bax mRNA水平没有变化，但Bcl-2 mRNA水平却显着下降（P <.05）。 p53 mRNA水平显着增加（P <.05），并与细胞凋亡率紧密匹配（R = 0.7，P <.05）。 caspase-3蛋白活性形式（细胞死亡的最终执行者）的表达趋于增加，并且与细胞凋亡率具有良好的相关性（R = 0.96，P <.01）。在体外培养颗粒细胞后，凋亡率倾向于在所有阶段增加，特别是在EF hCG阶段（P <.05）。 Bax和p53 mRNA趋于增加并显示出与凋亡率的良好相关性（R = 0.64，P <.05和0.86，P <.01）。 Bcl-2 mRNA水平在所有阶段都有下降的趋势，与细胞凋亡率无关。活性caspase-3蛋白的表达水平在各个阶段都有升高的趋势，并且与细胞凋亡率具有良好的相关性（R = 0.93，P <.01）。结论：根据卵泡发育阶段，大鼠卵巢颗粒细胞凋亡趋于增加。在线粒体依赖性基因中，p53与卵泡发育过程中的颗粒细胞凋亡密切相关。

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