经典Wnt/β-catenin信号通路介导子痫前期滋养细胞氧化应激损伤的机制研究  

Mechanism of classical Wnt/β-catenin signaling pathway mediated oxidaticve stress injury of trophoblast cells in preeclampsia

作　　者：罗欣[1] 庄白妹 饶海英[1] 李庆姝[2] 刘西茹[1] 漆洪波[1] 

机构地区：[1]重庆医科大学附属第一医院产科,重庆400016 [2]重庆医科大学基础医学院病理学教研室,重庆400016

出　　处：《重庆医科大学学报》2015年第6期801-805,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81300509;81300508;81471472)

摘　　要：目的:研究经典Wnt/β-连环蛋白(β-catenin)信号通路参与人滋养细胞氧化应激损伤的机制,探讨其在子痫前期(preeclampsia,PE)发病中的作用。方法:(1)采用免疫组化SP法和蛋白免疫印迹法(Western blot,WB)检测胎盘组织中β-catenin信号分子的表达。(2)将HTR8/SVneo分为正常培养对照组、缺氧/复氧(hypoxia/reoxygenation,H/R)处理组、氯化锂(Li Cl)+H/R处理组、Li Cl+正常培养组。利用WB法和间接免疫荧光法检测β-catenin在各组细胞中的表达。(3)利用流式细胞仪检测各组细胞凋亡指数及细胞内活性氧(reactive oxygen species,ROS)水平。(4)利用Transwell小室模型和明胶酶谱法分析检测各组HTR8/SVneo的侵袭率及基质金属蛋白酶(matrix metalloproteinase,MMP)-2、9的活性。结果:(1)与正常晚孕比较,子痫前期胎盘组织中β-catenin蛋白表达降低(P<0.05)。(2)与正常培养组相比较,H/R处理组的β-catenin蛋白水平显著下降,经Li Cl预处理后H/R细胞中的β-catenin蛋白表达增加(P<0.01)。(3)与正常对照组相比,H/R处理可导致HTR8/SVneo凋亡率及ROS水平增加(P<0.01);予以Li Cl预处理可有效抑制H/R导致的细胞凋亡及ROS聚集(P<0.05,P<0.01)。(4)H/R组细胞的体外侵袭能力及MMP-2、9的活性均低于正常培养组;经Li Cl预处理后,H/R组细胞的体外侵袭能力及清液中MMP-2、9的活性显著上升(P<0.01)。结论:Li Cl通过特异性激活Wnt/β-catenin信号通路,对滋养细胞氧化应激损伤有保护性作用。Objective：To investigate the roles of classic Wnt/β-catenin signaling pathway in the oxidative stress injury of trophoblasts involved in the development of preeclampsia. Methods：（1）Immunohistochemistry and Western blot were utilized to determine the expression of β-catenin in the placental tissues.（2）Cell treatment and classification of each group were as follow：normal culture group,H/R culture group,Li Cl＋normal culture group,Li Cl＋H/R culture group. The expressions and localization of β-catenin protein were detected by Western blot and immunofluorescence.（3）Flow cytometry assay was employed to verify the level of reactive oxygen species（ROS）and apoptosis index of HTR8/SVneo.（4）Changes of cell invasion rates were identified by transwell matrigel invasion assay;the activities of matrix metalloproteinase（MMP-2,9）were measured by gelatin zymography assay. Results：（1）The level of β-catenin protein decreased in placental tissues of preeclamptic compared with that in normal third trimester（P〈0.05）.（2）In HTR8/SVneo cells,β-catenin was located in both nucleus and cytoplasm. HTR8/SVneo cells exposed to H/R showed a significant decrease in β-catenin protein levels compared with the normal culture cells（P〈0.01）. Pretreated with Li Cl could upregulate the expression of β-catenin in H/R cells（P 〈0.01）.（3）Treatment with H/R could increase the level of ROS and apoptosis,and reduce the invasive ability of HTR8/SVneo cells in vitro（P 〈0.01）. Pretreated with Li Cl could inhibit the accumulation of ROS and apoptosis of cells caused by oxidative stress（P 〈0.01,P 〈0.05）.（4）The enzymatic activities of MMP-2,9 and the invasion rates of cells in vitro decreased significantly in HTR8/SVneo cells exposed to H/R（P〈0.01）,while pretreated with Li Cl could increase the enzymatic activities of MMP-2,9 and enhance the invasion rates of HTR8/SVneo cells exposed to H/R（P〈0.01）. Conclusion：Li Cl via activate the classical wnt/β-catenin signal

关 键 词：子痫前期 滋养细胞 氧化应激 Β-连环蛋白 氯化锂 

分 类 号：R714.24[医药卫生—妇产科学]