人胎盘源间充质干细胞条件培养液可上调缺氧状态下BeWo细胞活力和紧密连接因子的表达  

Human placenta-derived mesenchymal stem cell conditioned medium can upregulate BeWo cell viability and zonula occludens expression under hypoxia

作　　者：林清凡 解一新 陈婉清 叶振忠 陈幼芳 Lin Qingfan;Xie Yixin;Chen Wanqing;Ye Zhenzhong;Chen Youfang(Quanzhou Medical College,Quanzhou 362011,Fujian Province,China;Shishi General Hospital,Shishi 362700,Fujian Province,China;Fujian Genesis Stem Cell Co.,Ltd.,Quanzhou 362011,Fujian Province,China)

机构地区：[1]泉州医学高等专科学校,福建省泉州市362011 [2]石狮市(华侨)医院,福建省石狮市362700 [3]福建领航干细胞科技有限公司,福建省泉州市362011

出　　处：《中国组织工程研究》2021年第31期4970-4975,共6页Chinese Journal of Tissue Engineering Research

基　　金：福建省母婴健康服务应用技术协同创新中心基金(XJM1804),项目负责人:林清凡;福建省自然科学基金(2016J01422),项目负责人:陈幼芳;泉州市科技计划项目(2016Z005),项目参与人:林清凡。

摘　　要：背景:近年来间充质干细胞已是新药开发领域的研究热点,其旁分泌效应也已应用于多种组织损伤性细胞模型修复评估实验,但目前尚无研究探索其条件培养液是否对缺氧状态下人胎盘滋养层系细胞的紧密连接功能具有修复作用。目的:探讨人胎盘源间充质干细胞条件培养液对缺氧条件下人胎盘绒毛膜癌滋养层系BeWo细胞活力和紧密连接因子表达的影响。方法:采用1000μmol/L氯化钴处理12 h诱导BeWo细胞以构建胎盘屏障缺氧模型,检测细胞活力以及缺氧诱导因子1α和紧密连接因子闭合小环蛋白1、封闭蛋白4、封闭蛋白8的表达量变化;在缺氧模型中添加人胎盘源间充质干细胞条件培养液建立缺氧干预组、复氧修复组,检测细胞活力和上述4个基因的表达量改变;为探讨人胎盘源间充质干细胞条件培养液改变闭合小环蛋白1表达量的潜在机制,建立胰岛素样生长因子1缺氧干预组、胰岛素样生长因子1处理组,检测BeWo细胞活力以及闭合小环蛋白1表达量。结果与结论:①氯化钴诱导缺氧显著下调BeWo细胞活力,上调缺氧诱导因子1αmRNA和蛋白表达,下调闭合小环蛋白1和封闭蛋白4 mRNA和蛋白表达,上调封闭蛋白8 mRNA表达,但其蛋白表达下降;②人胎盘源间充质干细胞条件培养液明显提升缺氧条件下BeWo细胞活力和闭合小环蛋白1的表达,下调缺氧诱导因子1α的表达,但对封闭蛋白4无作用;③胰岛素样生长因子1干预缓解了缺氧对BeWo细胞活力的抑制作用,并上调闭合小环蛋白1的表达。BACKGROUND:In recent years,mesenchymal stem cells have become a research hotspot in the field of new drug development,and their paracrine effect has also been applied in the repair evaluation experiments of a variety of tissue injured cell models,but there is no research to explore whether their conditioned medium can repair the tight junction function of human placental trophoblast cells under hypoxia.OBJECTIVE:To demonstrate the effect of mesenchymal stem cell conditioned medium on cell viability and expression of tight junction factors in human placental choriocarcinoma trophoblast cell line BeWo under hypoxia.METHODS:BeWo cells were exposed to hypoxia induced by 1000μmol/L CoCl2 for 12 hours to construct the hypoxia model of placental barrier.For model evaluation,five indexes were measured,including cell viability and expression of hypoxia induced factors-1α,as well as three tight junction factors:zonula occludens 1,claudins 4 and claudins 8.Mesenchymal stem cell conditioned medium was added into hypoxia model to construct hypoxia intervention and reoxygenation groups.To investigate the potential mechanism of zonula occludens 1 expression recovery induced by mesenchymal stem cell conditioned medium,insulin-like growth factor 1 hypoxia intervention and treatment groups were established,and the BeWo cell viability and zonula occludens 1 expression were detected.RESULTS AND CONCLUSION:(1)BeWo cells viability,zonula occludens 1 and claudins 4 expression were reduced by CoCl2-mimic hypoxia,with hypoxia induced factors-1αmRNA and protein upregulation.However,the expression of claudins 8 mRNA was upregulated,and its protein was downregulated.(2)Human placenta-derived mesenchymal stem cell conditioned medium improved BeWo cells viability and recovered expression level of hypoxia induced factors-1αand zonula occludens 1 under hypoxia induced by CoCl2,but it has no effect on claudins 4.(3)Insulin like growth factor-1 intervention alleviated the inhibition of hypoxia on the viability of BeWo cells,and upregulated th

关 键 词：干细胞 人胎盘源间充质干细胞 条件培养液 氯化钴 缺氧 BEWO细胞 闭合小环蛋白1 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]