CAJ | 학술논문

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30 min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8 mRNA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Mr45 000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.
배경:태반화태막재모태액체평형과정중기착중요작용,이수천과태반화태막운수적분자화세포궤제목전상불십분청초,추측모태액체교환가능통과태반화태막상적수통도진행.목적:관찰인정상임신만기태반여태막조직중수통도단백8단백질수평적표체급기분포.설계、시간급지점:대비관찰,우2005-07/12재엄주의학원실험중심완성.재료:수집정상족월임신부궁분면적태반여태막조직양본각5례,산부년령(27±5)세,실험방법획의원윤리위원회비준.방법:태반태막조직재분면30 min내취양,용무균생리염수표세간정거제혈액,립즉방입액담중,전운지-80℃빙상보존비용.주요관찰지표:운용반전록-취합매련반응법、면역조직화학법급Western인적법검측수통도단백8재태반여태막조직중적표체여분포.결과:반전록-취합매련반응현시,수통도단백8 mRNA재태반화태막조직중균유표체.수통도단백8 Western인적검측태반화태막조직현시,일특이성조대재Mr45 000좌우.면역조직화학결과현시,수통도단백8표체우태반합체자양세포、양막상피세포급평활융모막세포자양세포.결론:재단백질수평현시,수통도단백8표체우태반적합체자양세포、양막상피세포급평활융모막세포자양세포.
BACKGROUND: Placenta and fetal membrane play an important role In maternal-fetal homeostasis. However, the molecular and cellular mechanisms underlying water transfer across placenta and amniotic membrane remain unknown. It is hypothesized that maternal-fetal fluid exchanges via aquaporin (AQP) water channels in the placenta and fetal membrane.OBJECTIVE: To investigate AQP8 protein expression in normal human placenta and fetal membrane.DESIGN, TIME AND SETTING: A control observation was performed at the Central Laboratory of Guangzhou Medical College from July to December 2005.MATERIALS: Human placenta and fetal membrane tissues from 5 elective cesarean section deliveries of normal term pregnancies (range 37-42 weeks) were studied. Maternal age averaged (27?) years old. Experimental protocol was approved by the Hospital's Ethics Committee.METHODS: Thirty minutes after delivery, fetal membrane and placenta were dissected and washed with sterile physiological saline. Some were frozen at -80?, and the remaining tissues were fixed for 24-48 hours with 10% neutral formalin and paraffin embedded for immunohistochemical staining.MAIN OUTCOME MEASURES: AQP8 expression and distribution in human placenta and fetal membrane were detected by the reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blotting analysis.RESULTS: RT-PCR results showed that AQP8 mRNA was expressed in both placenta and fetal membrane tissues. Western blotting analysis also yielded positive results in placenta and fetal membrane with a specific band site at approximately 45 000.Immunohistochemistry results revealed that AQP8 protein was expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.CONCLUSION: At protein level, AQP8 is expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.