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背景:胎盘和胎膜在母胎液体平衡过程中起着重要作用,而水穿过胎盘和胎膜运输的分子和细胞机制目前尚不十分清楚,推测母胎液体交换可能通过胎盘和胎膜上的水通道进行。目的:观察人正常妊娠晚期胎盘与胎膜组织中水通道蛋白8蛋白质水平的表达及其分布。设计、时间及地点:对比观察,于2005-07/12在广州医学院实验中心完成。材料:收集正常足月妊娠剖宫分娩的胎盘与胎膜组织样本各5例,产妇年龄(27±5)岁,实验方法获医院伦理委员会批准。方法:胎盘胎膜组织在分娩30min内取样,用无菌生理盐水漂洗干净去除血液,立即放入液氮中,转运至-80℃冰箱保存备用。主要观察指标:运用反转录-聚合酶链反应法、免疫组织化学法及Western印迹法检测水通道蛋白8在胎盘与胎膜组织中的表达与分布。结果:反转录-聚合酶链反应显示,水通道蛋白8mRNA在胎盘和胎膜组织中均有表达。水通道蛋白8 Western印迹检测胎盘和胎膜组织显示,一特异性条带在Mr45000左右。免疫组织化学结果显示,水通道蛋白8表达于胎盘合体滋养细胞、羊膜上皮细胞及平滑绒毛膜细胞滋养细胞。结论:在蛋白质水平显示,水通道蛋白8表达于胎盘的合体滋养细胞、羊膜上皮细胞及平滑绒毛膜细胞滋养细胞。
BACKGROUND: The placenta and fetal membranes play an important role in the fluid balance of the fetal fluid. The molecular and cellular mechanisms by which water travels across the placenta and fetal membranes are not well understood. It is speculated that fluid exchange through the placenta and fetal membranes may be presumed Channel. Objective: To observe the expression and distribution of aquaporin-8 protein in placenta and fetal membranes in normal late pregnancy. DESIGN, TIME AND SETTING: The comparative observation was performed at the Experimental Center of Guangzhou Medical College between July 2005 and December 2005. MATERIALS: A total of 5 placenta and fetal membranes samples were collected from cesarean delivery in normal term pregnancy. The age of the patients was (27 ± 5) years old. The experimental method was approved by the Hospital Ethics Committee. Methods: Placenta fetal tissue samples within 30min of delivery, rinsed with sterile saline to remove the blood, immediately into the liquid nitrogen, transported to -80 ℃ refrigerator for later use. MAIN OUTCOME MEASURES: The expression and distribution of aquaporin 8 in placenta and fetal membranes were detected by reverse transcription - polymerase chain reaction, immunohistochemistry and Western blotting. Results: Reverse transcription - polymerase chain reaction showed that aquaporin 8 mRNA was expressed in both placenta and fetal membranes. Aquaporin 8 Western Blotting Placenta and fetal membranes revealed that a specific band around Mr45000. Immunohistochemistry showed that aquaporin 8 was expressed in placental syncytiotrophoblasts, amniotic epithelial cells and placental chorionic cytoplastic cells. CONCLUSIONS: At the protein level, aquaporin 8 is expressed on placental syncytiotrophoblasts, amniotic epithelial cells, and placental chorionic cytoplastic cells.