双炔失碳酯抗小鼠早孕的形态学观察

作者用双炔失碳酯3mg/kg注入妊娠第6天小鼠腹腔内,于妊娠第12天处死作病理观察。见:胚体坏死;子宫蜕膜及滋养层细胞出现不同程度退变坏死,部份为水样变性,细胞肿胀,胞浆内出现巨大的空泡,胞质无内容,另一部份为凝固性坏死,细胞轮廓呈嗜伊红残影;卵巢颗粒黄体细胞出现不同程度的核固缩、细胞质呈现空泡,出现胞质溶解现象。本实验结果为探讨该药抗孕机理提供了形态学资料。     

Co-culture of early embryo with human decidual stromal cellsin vitro by improvement of early embryo development

Summary An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM + 0.4 % bovine serum albumin (BSA) and 163 embryos cultured in MEM + 15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73 % developed to the morula stage and 67.21 % cavitated to blastocysts with 59.74 % hatching, as compared with 61.34 % to morula stage, 48.47 % to blastocysts and none hatching in the controls, respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.     

p38丝裂原活化蛋白激酶特异性抑制剂对小鼠早胚发育及植入的影响

目的观察p38丝裂原活化蛋白激酶（p38MAPK）特异性抑制剂SB203580对小鼠早胚发育及植入的影响。方法①采用体外胚胎培养技术,将孕2?d小鼠早胚随机分组,接种到添加不同浓度抑制剂的培养液内进行培养,以观察p38MAPK特异性抑制剂对早胚发育的影响;②体内宫腔注射：取孕4?d小鼠,由子宫角注射不同浓度的SB203580,于孕8?d检查胚胎植入数,同时取子宫,观察内膜的组织学变化。结果①添加抑制剂组的早胚不能发育为胚泡,停滞在8～16细胞阶段。但去除抑制剂后继续培养,幼胚仍能发育到胚泡阶段;②注射SB203580组的小鼠,胚胎植入数明显少于对照组（P＜0.01）。镜下可见,其胚胎组织呈退化坏死状态,胚胎周围蜕膜细胞呈明显的空泡样变性,蜕膜及胚胎中均见大量血细胞浸润。结论①p38MAPK在小鼠植入前胚胎发育过程中起作用;②p38MAPK特异性抑制剂SB203580可抑制在体小鼠胚泡植入及植入后胚胎和蜕膜细胞的发育。     

被动吸烟对大鼠卵巢组织结构的影响

目的 利用Wistar大鼠被动吸烟模型,观察被动吸烟对Wistar大鼠卵巢结构的影响,为提倡生育期妇女避免被动吸烟提供新的理论依据.方法 建立大鼠烟雾吸入模型.32只健康雌性Wistar大鼠,每只体重200～ 250g,随机分为实验组和对照组,每组16只.实验组被动吸烟3个月,对照组正常饲养.两组大鼠均于饲养3个月后处死,取双侧卵巢.常规石蜡切片,HE染色,光镜观察卵巢形态结构;以半薄切片为指导进行超薄切片,醋酸双氧铀染色,透射电镜观察卵巢超微结构.结果 ①光镜观察：吸烟组大鼠卵巢充血性囊性退变,皮质萎缩,卵泡颗粒细胞排列紊乱;生长卵泡的颗粒细胞松散变性,排列不整齐;卵泡数减少,成熟的卵母细胞数目明显减少;髓质血管收缩,减少;间质疏松.②电镜观察;对照组可见多个卵泡细胞,细胞内有空白颗粒及高密度颗粒.实验组可见多个颗粒细胞层,内有增粗的内质网、核糖体;核周隙局部增宽,线粒体模糊;线粒体水样变、空泡样变,线粒体嵴断裂、模糊;内质网脱颗粒样变.结论 ①成功建立了大鼠被动吸烟模型.②被动吸烟可破坏雌鼠卵巢的组织结构和超微结构,为吸烟对女性生殖功能的影响提供形态学基础.     

Co-Culture of Early Embryo with Human Decidual Stromal Cells in vitro by Improvement of Early Embryo Development

The quality of the in vitro culture conditionsfor human pre--implantation embryos is one of themost critical aspects of successful in vitro fertilization and embryo transfer (IVF--ET) and other biology techniques, such as embryo clone, geneknock--out, pre--implantation genetic diagnosis etc.The culture of pre--im:7lantation mammalian embryos can take place in a range of embryo culturemedia, ranging from simple chemically defined media to more complex media. In vitro pre--implantation embryo …     

米非司酮终止早孕后对再次妊娠蜕膜ER、PR的影响

目的 :探讨米非司酮配伍米索前列醇 (米索 )中止早孕后对再次妊娠早孕蜕膜雌激素受体 (ER)、孕激素受体 (PR)的影响。方法 :取停经 4 9d内、无口服米非司酮史妇女早孕蜕膜绒毛 2 0例 ,分为两组 (Ⅰ、Ⅱ组 )。另取有使用米非司酮抗早孕史再孕妇女蜕膜绒毛 2 0例 ,分为两组 (Ⅲ、Ⅳ组 )。其中Ⅰ、Ⅲ组行人工流产 ,Ⅱ、Ⅳ组行药物流产。绒毛组织均行HE染色光镜下观察 ,蜕膜组织应用免疫组织化学方法及电子计算机图像分析技术观察ER、PR的变化。结果 :①Ⅰ、Ⅲ组合体滋养细胞及细胞滋养细胞无水肿变性 ,间充质无异常染色。Ⅱ、Ⅳ组绒毛水肿变性明显。②早孕蜕膜ER、PR均位于大部分间质细胞及少数腺上皮细胞的胞核内。 4组受体水平积分Ⅰ、Ⅱ组间有显著差异 (P <0 .0 1) ;Ⅲ、Ⅳ组间有显著差异 (P <0 .0 1) ;Ⅰ、Ⅲ组间及Ⅱ、Ⅳ组间均无显著差异 (P >0 .0 5 )。结论 :①米非司酮配伍米索可引起早孕绒毛水肿变性坏死 ;②米非司酮配伍米索可使早孕蜕膜ER、PR水平下降 ,但月经恢复后ER、PR水平已回复至正常 ,再次妊娠后蜕膜ER、PR与未服用米非司酮及米索者早孕蜕膜ER、PR无差异。前次药物流产效果正常者 ,再次行药物流产同样有效     

Effects of Mifepristone （RU486） on Development and Ultrastructure of Mouse Blastocysts

Objective To investigate effects of mifepristone （MP） on the development and ultrastructure of mouse blastocysts in vivo Methods Sixty female mice were equally divided into 4 groups： control group （group A）, 1.9 mg/kg MP group （group B）, 5.6 mg/kg MP group （group C）, and 16.8 mg/kg MP group （group D）. The female mice of 4 groups undertook a superovulation method. The development of obtained blastocysts was evaluated, and ultrastructural changes of the blastocysts were observed by transmission electron microscope. Results In comparison with group A, the development rate of blastocysts in group B showed no difference （P〉0.05）, while the development rate of blastocysts in both group C and group D was significantly lower （P〈0.05）. With doses of 5.6 mg/kg and 16.8 mg/kg, the blastocysts showed granular appearance of the cytoplasm, irregular cell borders, enlarged perivitelline space and degeneration. Ultrastructure of the blastocysts in group B was similar to group A, except a little number of fat droplets in the cytoplasm. In group C, the microvilli on apical surface was decreased in number or even disappeared, mitochondria were under developed, a lot of filamentous crystals were found in the cytoplasm. Cellular junctions were defected. In group D, the blastocyst cells were irregular in shape, mitochondria were frequently vacuolated, the nucleolus was enlarged, nuclear membrane was ruptured, and chromatin was slack. Conclusion MP could damage to the ultrastructure of mouse blastocyst, and was responsible for the inhibition of blastocyst development. The inhibitory effect of MP would be in a dose-dependent fashion.     

雌激素、孕激素对小鼠胚卵体外发育及胚泡着床的影响

目的:研究雌、孕激素在小鼠胚卵体外发育及胚泡着床过程中的作用.方法:在小鼠桑椹胚与人蜕膜细胞共培养体系中分别加入雌激素(E2组)、孕激素(P组)、雌激素和孕激素(E2 P4组),观察各组胚卵发育以及胚泡粘附和铺展情况.结果:E2组桑椹胚向胚泡的发育率高于对照组,E2 P4组胚泡的粘附率和滋养层细胞的铺展率均较对照组高.结论:单独用雌激素能够促进小鼠桑椹胚发育成胚泡,雌激素和孕激素联合应用能够促进胚泡的粘附和铺展.     

超排卵对体外培养胚胎IL-6分泌的影响

【目的】探讨超排卵对小鼠体外培养胚胎发育及其分泌IL-6的影响。【方法】采用超排后自然受孕和正常自然受孕ICR小鼠各50只,收集受精卵体外培养3 d,观察胚胎发育状态。应用酶联免疫吸附法检测各胚胎培养液滴IL-6蛋白的浓度。【结果】超排组囊胚形成率明显低于正常组(P<0.05),但碎片和阻滞胚胎率高于对照组(P<0.05)。ELISA结果显示超排组IL-6分泌显著低于正常组(P<0.01)。【结论】控制性超排卵能够降低IL-6分泌,可能与其影响胚胎发育有关。     

超声波对小鼠胚胎表达整合素及其着床能力的影响

目的：研究超声波照射对小胎胚表达整合素α1、α5、β1、β3的影响充及与其着床能力的关系。方法：取小鼠二细胞胚经超声波照射后体外培养至囊胚期,用免疫组化法检测其整合素α1、α5、β1、β3的表达及其着床能力的改变,并与未经照射的正常胚胎比较,结果：1、超声波照射组胚贴附率,滋养细胞外延生长率均显著低于正常胚胎。2、正常胚胎桑椹胚期即有α5、β1、β3整合素表达,而无α1表达：囊胚期α5、β1、β3表达增强,仍无α1表达：α1只出现在外延生长的滋养细胞。3、超声波照射组胚胎β3整合素的表达明显弱于正常组。且未见α1表达,而α5、β1的表达两组间无差异。结论：超声波照射使胚胎着床能力明显降低。可能与其影响胚胎细胞α1、3整合素的正常表达有关。