摘要
为探讨γ-分泌酶抑制剂(DAPT)阻断Notch信号通路对BMSCs分化肺泡上皮细胞过程中的影响。本研究采用BMSCs与MLE-12非接触共培养,并在共培养中加入DAPT,实验分3组:空白对照组、共培养组、DAPT共培养组。共培养10 d后用光镜观察BMSCs的形态结构变化,Western blotting和RT-qPCR检测Notch信号通路相关基因Notch1,Ⅱ型肺泡上皮细胞特异性表面活性蛋白(surfactant protein C,SPC)、Ⅰ型肺泡上皮细胞标志水通道蛋白(aquaporin 5,AQP5)的表达。结果表明共培养10 d后的BMSCs变为似铺路石样上皮细胞形态;和空白对照组相比,共培养组、DAPT共培养组中Notch1、SPC、AQP5蛋白及mRNA表达升高(p<0.05);与共培养组相比,DAPT共培养组中Notch1、SPC、AQP5蛋白及mRNA的表达减少(p<0.05)。因此推测BMSCs在MLE-12诱导下可以定向分化为肺泡上皮细胞,且分化的过程中Notch信号通路被激活,DAPT阻断Notch信号通路能抑制BMSCs分化为肺泡上皮细胞。
In order to explore the effect of γ-secretase inhibitor blocking Notch signaling pathway on bone mesenchymal stem cells differentiated into alveolar epithelial cells, the BMSCs and MLE-12 were used for non contact co-culture, and DAPT was added into the co-culture. The experiments were divided into 3 groups： the blank control group, co-culture group and DAPT co-culture group. The morphological changes of BMSCs were observed by light microscope after 10 days of co-culture, Western blotting and RT-qPCR were used to detect Notch signaling pathway related genes ofNotchl, the expression of type II alveolar epithelial cell specific surface active protein （surfactant protein C, SPC） and type I alveolar epithelial cell marker water channel protein （aquaporin 5, AQP5）. Results indicated that BMSCs were changed into the shape of stone like epithelial cells after 10 days of co-culture. Compared with the blank control group, the expression ofNotchl, SPC, AQP5 and mRNA were increased in the co-culture group and DAPT co-culture group （p〈0.05）. Compared with co-culture group, the expression of Notchl, SPC, AQP5 and mRNA in DAPT co-culture group wee decreased （p〈0.05）. Therefore, we can infer that BMSCs can be induced to differentiate into alveolar epithelial cells under MLE-12 induction and Notch signaling pathway was activated during differentiation. DAPT blocking Notch signaling pathway may inhibit BMSCs differentiated into alveolar epithelial cells.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第5期1727-1731,共5页
Genomics and Applied Biology
基金
国家自然基金(81660009)
江西省自然基金重大项目(20161ACB20012)共同资助
关键词
骨髓间充质干细胞
NOTCH信号通路
肺泡上皮细胞
分化
Bone mesenchymal stem cells, Notch signaling pathway, Alveolar epithelial cell, Differentiation
