钠-钾ATP酶介导间充质干细胞修复大鼠急性肺损伤  

Mesenchymal stem cells ameliorated aucte lung injury via Na-K-ATPase of alveolar type Ⅱ cells in rats

作　　者：李基伟[1] 魏立[1] 务森[1] 陈晓[1] 张宁[1] 朱晓明[1] 陈重[1] 

机构地区：[1]河南省人民医院胸外二科,郑州450003

出　　处：《中华实验外科杂志》2017年第8期1345-1347,共3页Chinese Journal of Experimental Surgery

基　　金：河南省人民医院“23456人才工程”科研专项基金

摘　　要：目的 探讨间充质干细胞（MSCs）旁分泌作用修复急性肺损伤（ALI）的机制.方法 实验设阴性组、炎症组、修复组及对照组,每组大鼠分别注射磷酸盐缓冲液（PBS）、脂多糖（LPS）、LPS＋MSCs和MSCs,72h后测定肺湿/干比及制作苏木素-伊红（HE）病理切片.体外共培养大鼠肺泡Ⅱ型上皮细胞（AT-Ⅱ）及MSCs,设置阴性组、炎症组、修复组及对照组,共培养72h后提取大鼠AT-Ⅱ蛋白及mRNA,Western blot及定量聚合酶链反应方法观察Na-K-ATPaseα1亚基在蛋白及基因水平的改变以及测定AT-Ⅱ的Na-K-ATPase活性.结果 LPS明显造成大鼠ALI,鼠尾静脉注入MSCs可明显缓解炎症损伤;阴性组、炎症组、修复组及对照组肺湿干比分别为：4.33±0.24、5.21±0.21、4.34±0.19和4.44±0.14,MSCs可明显降低肺湿/干比（P=0.000）.体外实验证实MSCs可明显提高AT-Ⅱ的Na-K-ATPase活性.阴性组、炎症组、修复组及对照组Na-K-ATPase的活性分别为0.54±0.01、0.68±0.02、0.75±0.01和0.62±0.00,炎症环境可提高Na-K-ATPase活性,P=0.000;MSCs明显提高Na-K-ATPase活性,P=0.000.4组α1亚基蛋白表达分别为0.737±0.039、0.610±0.022、0.780±0.029 和0.838±0.022,MSCs明显提高Na-K-ATPaseα1亚基的蛋白表达,P=0.000.MSCs同时促进α1基因的表达,4组mRNA表达分别为1.000±0.000、0.787±0.044、3.960±0.091及4.466±0.071,P=0.000.结论 MSCs不仅提高AT-Ⅱ的Na-K-ATPaseα1亚基在蛋白和基因的表达,同时提高Na-K-ATPase活性,从而缓解ALI.Objective To study the mechanism by which acute lung injury was ameliorated by mesenchymal stem cells （MSCs） in rats.Methods Rats were injected with phosphate buffered saline （PBS）,lipopolysaccharide （LPS）,LPS＋MSCs or MSCs,respectively.Histopathology and wet/dry analysis in four groups was performed.Therapeutic effects were further evaluated in vitro after co-culture.The expression of α1 subunit of Na-K-ATPase in alveolar type Ⅱ cells （AT-Ⅱ） was detected by Western blotting and quantitative polymerase chain reaction （qPCR） at protein and gene levels respectively.Results LPS injection resulted in marked inflammatory injury according to hematoxylin and eosin （HE）-stained lung sections.MSC administration reduced the lung injury,and the wet/dry ratio in four different groups was 4.33±0.24,5.21±0.21,4.34±0.19 and 4.44±0.14 respectively.LPS administration to rats resulted in increased ratio of wet/dry versus rats injected with PBS （P=0.000）.The decrease in wet/dry ratio was lower as LPS-exposed rats were treated with MSCs （P=0.000）.In addition,the protein expression of α1 subunit was 0.737±0.039,0.610±0.022,0.780±0.029 and 0.838±0.022 respectively.Meanwhile,the mRNA expression of α1 subunit was 1.000±0.000,0.787±0.044,3.960±0.091 and 4.466±0.071 respectively.As shown,the protein expression of α1 subunit was decreased after inflammatory insult （P=0.000）.Co-culture with MSCs increased the expression of α1 subunit （P=0.000）.Na-K-ATPase activity was 0.54±0.01,0.68±0.02,0.75±0.01 and 0.62±0.00 respectiely.It was not only up-regulated at inflammatory situation but also further increased by MSCs.Conclusion MSCs provided therapeutic benefits via up-regulating the expression of α1 subunit and increasing activity of Na＋/K＋-ATPase in AT-Ⅱ cells.

关 键 词：间充质干细胞 肺泡Ⅱ型上皮细胞 NA-K-ATPASE 

分 类 号：R563.8[医药卫生—呼吸系统]