激活素A及卵泡抑素对冻融人胎儿卵巢雌二醇分泌影响.docVIP

激活素A及卵泡抑素对冻融人胎儿卵巢雌二醇分泌影响 　 作者：池海虹，周颖，祝怀平，金畅，叶碧绿 【关键词】 激活素A；卵泡抑素；胎儿；卵巢；雌二醇 Abstract: Objective:To investigate the effect of Activin A-follistatin on steroidogenesis of frozen-thawed human fetal ovary in vitro. Methods: The frozen-thawed fetal ovarian tissue was cultured individually in minimal essential medium (MEM) for 8 days. Under the experimental conditions, the medium was supplemented with either Activin-A (Act A group) or follistatin (FS group) or the combinations of Activin-A and follistatin (A+F group). The concentration of E2 in cultured medium was detected with immunoluminescence method. Results: The levels of E2 at 4th day, 6th day and 8th day were higher than that at 2nd day (Plt;0.05), The secretion of E2 increased at first, then decreased with the highest level at 6th day. The level of E2 of Act A group was significantly greater than that of the control group, FS group and A+F group (Plt;0.01). Conclusion: Frozen-thawed human fetal ovary can excrete estrodiol under the condition of tissue culture. Act A can increase the excretion of estrodiol in cultivated ovarian tissue, and the effection of Act A can be suppressed by FS. Key words: activin A; follistatin; fetal; ovarian tissue; estrodiol 近年来发现的一组性腺起源的细胞因子--激活素(Activins)，对卵泡发育有着重要的局部调节作用，尤其是激活素A(Activin A，Act A)。尽管有较多的研究表明了激活素对不同动物卵泡发育的影响[1-3]，但对人胎儿卵巢内卵泡发育的影响尚未见报道。本研究以胎儿卵巢为研究对象，比较Act A和卵泡抑素(FS)在体外组织培养条件下对冻融人胎儿卵巢E2分泌的影响，探讨其在人卵泡发生过程中可能的调节作用。 1 材料和方法 1.1 标本 卵巢组织取自因计划生育或医疗原因需行药物引产（利凡诺＋米菲司酮）的32周胎儿（温州医学院教学医院分娩室提供）共3例，于引产后2 h内无菌条件下取材，置于预冷的Leiboviz-15(L-15，美国sigma公司产品)基础培养液中待用。本研究经我院伦理委员会批准。 1.2 试剂和仪器 基础培养液为MEM培养液(购自GIBCO公司)，含5%小牛血清，100 U/ml的FSH, 100 IU/ml青霉素和100 IU/ml链霉素，1/100 ITS(胰岛素-转铁蛋白-硒，insulin-transferrin-selenium)，pH 7.0～7.2。Act A、FSH、FS均为Sigma公司产品。E2测定采用美国贝克曼库而特公司BXI800仪器及试剂。 1.2 实验方法 1.2.1 冷冻及复苏: 将胎儿卵巢组织去除髓质及周围结缔组织后，切成5 mm×5 mm×1 mm大小，放入装有冷冻保护剂1.5 mol/L丙二醇（PROH）的冷冻管中，按照Oktay[4]提出的慢冻速融法处理。 1.2.2 体外培养及分组:冻融卵巢皮质块切成2 mm×2 mm×1 mm大小，随机放入加有0.8 ml培养液的24孔培养板内，37 ℃、5%CO2培养箱内培养，隔天半量换液，共培养8 d。实验共分四组：M