间充质干细胞条件培养基对肺泡上皮液体清除能力的影响

摘要

目的 探讨小鼠骨髓间充质干细胞(BMSCs)条件培养基对小鼠肺泡液体清除的影响和对人肺泡上皮细胞生存能力的作用,并明确小鼠BMSCs条件培养基在肺脏液体清除中的作用机制.方法 细胞贴壁法分离培养小鼠BMSCs,应用流式细胞术对小鼠BMSCs的表面标记物进行鉴定;运用酶标仪测定小牛血清白蛋白浓度的方法测定小鼠在体肺泡液体清除率;利用活细胞计数盒(CCK-8)试剂研究小鼠BMSCs条件培养基对人H441肺泡上皮细胞生存能力的影响;采用蛋白质印迹法研究小鼠BMSCs条件培养基对人肺泡上皮细胞钠通道蛋白水平的影响.结果 小鼠BMSCs稳定表达CD44,阴性表达CD34;小鼠BMSCs条件培养基能够提升在体小鼠肺泡液体清除率;小鼠BMSCs的条件培养基能够提高人肺泡上皮细胞的生存能力;蛋白质印迹法结果显示小鼠BMSCs条件培养基能够增加人肺泡上皮细胞钠通道的蛋白水平表达.结论 小鼠BMSCs条件培养基能够增强肺泡上皮细胞的液体清除并能改善其生存能力,机制可能与肺泡上皮细胞钠通道蛋白表达的增加有关.%Objective To explore the effects of conditioned media from mice bone marrow mesenchymal stem cells (BMSCs) on the mice alveolar fluid clearance (AFC) and the viability of human alveolar epithelial H441 cells,and to investigate the potential mechanisms of the former.Methods Mice BMSCs were isolated and cultured through cell adherence method,and they were identified with phenotypic markers by flow cytometry.AFC was measured using bovine serum albumin protein assays.A CCK-8 kit was used to investigate the effects of conditioned media from mice BMSCs on the viability of human alveolar epithelial H441 cell.Western blotting was applied to analyze the effects of conditioned media from mice BMSCs on the sodium channels protein levels of human alveolar epithelial cells.Results Mice BMSCs positively expressed the cell surface markers CD44 and negatively expressed CD34 by flow cytometry.Conditioned media from mice BMSCs could increase the rate ofin vivo AFC in mice.Conditioned media from mice BMSCs could increase the viability of human alveolar epithelial cells.Western blotting showed that the expression of sodium channel in human alveolar epithelial cells was significantly elevated by conditioned media from mice BMSCs.Conclusion Conditioned media from mice BMSCs can increase AFC and the viability of alveolar epithelial cell,and the former may be related to the augment of epithelial sodium channel protein expression.