摘要
该研究旨在探讨小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)通过miR-130b对上皮钠通道(epithelial sodium channel,ENaC)的影响。将分离与培养的小鼠BMSCs接种到Transwell小室中,然后与H441细胞进行共培养。利用CCK-8试剂盒检测BMSCs对H441细胞生存能力的影响;采用Western blot技术检测BMSCs对共培养的H441细胞中γ-ENaC蛋白水平的影响;qRT-PCR技术检测与BMSCs共培养的H441细胞中miR-130b表达情况,然后将此microRNA转染到普通培养的H441细胞中,在蛋白水平进一步验证其对H441细胞中γ-ENaC的影响。实验结果表明,BMSCs能够增强H441细胞的生存能力;同时BMSCs能分别增加共培养的H441细胞中γ-ENaC的蛋白水平以及miR-130b的转录水平;Western blot实验进一步证实,miR-130b转染至H441细胞后能够增加其γ-ENaC的蛋白表达。由此我们推测,BMSCs能够增强H441细胞的生存能力并且可能通过miR-130b发挥其对γ-ENaC的蛋白水平调控作用。
The aim of this study was to investigate the effects of mouse BMSCs(bone marrow mesenchymal stem cells)on ENaC(epithelial sodium channel)via miR-130b.The isolated and cultured mouse BMSCs were seeded onto the transwell inserts and then co-cultured with H441 cells.The effect of BMSCs on the viability of H441 cells was detected by CCK-8 reagent.The effect of BMSCs on the level ofγ-ENaC protein in co-cultured H441 cells was detected by Western blot.The expression of miR-130b in H441 cells co-cultured with BMSCs was detected by qRT-PCR.The microRNA was then transfected into normal cultured H441 cells,and its effect onγ-ENaC in H441 cells was further verified at the protein level.The results showed that BMSCs could enhance the viability of H441 cells.BMSCs could increase the protein level ofγ-ENaC and the transcription level of miR-130b in cocultured H441 cells respectively.Western blot analysis further confirmed that miR-130b could increase the protein expression ofγ-ENaC after transfection into H441 cells.Thus,we speculate that BMSCs can enhance the viability of H441 cells and may exert its regulation ofγ-ENaC protein level through miR-130b.
作者
张红蕾
周祉妤
候亚鹏
刘宏飞
王琳
丁炎
聂宏光
ZHANG Honglei;ZHOU Zhiyu;HOU Yapeng;LIU Hongfei;WANG Lin;DING Yan;NIE Hongguang(Department of Stem Cells and Regenerative Medicine,China Medical University,Shenyang 110122,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2020年第1期88-93,共6页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81670010)
辽宁省重点研发计划指导计划项目(批准号:2018225077)资助的课题。
