水通道蛋白3在人羊膜上皮细胞中的表达及调控  

Expression and regulation of aquaporin 3 in human amnion epithelial cells

作　　者：马小燕[1] 沈奇[1] 王晶晶[1] 谢爱兰[1] 朱雪琼[1] 

机构地区：[1]温州医学院附属第二医院妇产科,325027

出　　处：《中华围产医学杂志》2013年第5期288-293,共6页Chinese Journal of Perinatal Medicine

基　　金：浙江省中医药重点研究计划（2010ZZ009）;温州市科技合作计划项目（H20100067）

摘　　要：目的研究丝裂原活化蛋白激酶（mitogen-activated protein kinases，MAPK）／细胞外信号调节激酶（extracellular signal regulated kinase，ERK）信号转导通路对人羊膜上皮细胞中水通道蛋白3（aquaporin，AQP3）表达的调控作用。方法选择2011年1月至11月，在温州医学院附属第二医院产科分娩的不明原因羊水过少和羊水量正常的足月单胎妊娠产妇各10例，均为剖宫产分娩。原代培养羊膜上皮细胞，并用ERK1／2抑制剂U0126进行处理。浓度为0、5、10、20和40umol／L的U0126分别作用12h；再选取使磷酸化ERK1／2表达水平最低的浓度为最佳浓度，作用于细胞0、2、6、12和24h。采用免疫细胞化学染色检测AQP3蛋白定位，Western印迹检测总ERK1／2、磷酸化ERK1／2和AQP3蛋白表达水平。统计学方法采用t检验和方差分析。结果（1）羊水过少组磷酸化ERK1／2和AQP3的表达均低于羊水量正常组（ERK1／2：2．46±0．25与3．46±0．33；AQP3：1．56±0．10与2．34±0．18，t=9．243和13．292，P〈0．01）。（2）羊水过少组中U0126不同浓度组间比较，总ERK1／2表达差异无统计学意义（F=0．365，P〉0．05）；5、10、20、40umol／L组磷酸化ERK1／2和AQP3表达均低于0μmol／L组（磷酸化ERK1／2：0．96±0．16、1．12±0．13、0．98±0．17、1．02±0．26与2．46±0．25；AQP3：1．10±0．09、1．12±0．08、1．13±0．06、1．11±0．06与1．56士0．10，P〈0．05）；但5μmol／L组与10、20和40μmol／L组比较，差异均无统计学意义（P〉0．05）。羊水过少组U0126作用的最佳浓度为5μmol／L。该浓度作用2h组磷酸化ERK1／2和AQP3表达均低于0h组（磷酸化ERK1／2：1．27±0．29与2．55±0．12；AQP3：1．44±o．12与2．15±0．09，P〈0．05），但2h组与6、12和24h组比较，差异均无统计学意义（P〉0．05）。最佳作用时间为2h。（3）羊水量正常组和羊水过少组产妇的羊膜上皮细胞中，�Objective To investigate the role of mitogen-activated protein kinases （MAPK）/ extracellular signal regulated kinasel/2 （ERK1/2） signal transduction pathway in regulating the expression of aquaporin 3 （AQP3） in human amnion epithelial cells. Methods Primary cell culture of human amnion epithelial ceils deriving from amnion of term pregnancy with normal amniotic fluid volume （AFV） or isolated oligohydramnios was conducted in the Second Affiliated Hospital of Wenzhou Medical College from January to November 2011. Either group included 10 elective cesarean cases. The primarily cultured cells were treated with different concentrations （0, 5, 10, 20 and 40 μmol/L） of ERK inhibitors （U0126） for 12 h, and then the optimal concentration of U0126 which resulted in the lowest expression of phosphor-ERK1/2 （p-ERK1/2） was added for different durations （0, 2, 6, 12 and 24 h）. Immunocytochemistry was used to detect the localization of AQP3 and Western blot analysis was used to examine the expression of total ERK1/2, p ERK1/2 and AQP3 in human amnion epithelial cells. Statistical analysis was performed by t-test and one-way ANOVA. Results （1） Compared with those in normal AFV group, p-ERK1/2 and AQP3 expression in human amnion epithelium cells decreased in oligohydramnios group, respectively （p-ERK1/2：3.46 ± 0.33 and 2.46±0.25;AQP3：2.34±0.18 and 1.56±0.10, t= 9.243 and 13.292, P〈0.01）. （2） In oligohydramnios groups, after treated with different concentrations of U0126, the expressions of total ERK1/2 did not change （F=0.365, P〉0.05）. The expression of p-ERK1/2 and AQP3 in 5, 10, 20, 40 μmol/L U0126 （p-ERK1/2： 0. 96±0.16, 1.12±0.13, 0.98±0.17 and 1.02±0.26; AQP3： 1.10±0.09, 1.12±0.08, 1.13±0.06 and 1.11±0.06） were all significantly lower than those in 0 μmol/L U0126 group （p-ERK1/2： 2.46±0.25; AQP3： 1.56±0.10, P〈0.05）. However, the expression of p-ERK1/2 and AQP3 showed no significant difference among 5, 10, 20, and 40/lmol/L U0126 groups

关 键 词：水通道蛋白质3 细胞外信号调节MAP激酶类 羊膜 上皮细胞 羊水过少 

分 类 号：R714[医药卫生—妇产科学]