骨髓间充质干细胞旁分泌KGF对LPS诱导小鼠肺上皮细胞损伤的修复作用  

REPAIR EFFECT OF KERATINOCYTE GROWTH FACTOR SECRETED BY BONE MARROW MESENCHYMAL STEM CELLS THROUGH A PARACRINE MECHANISM ON LIPOPOLYSACCHARIDE-INDUCED MOUSE LUNG EPITHELIAL CELL DAMAGE

作　　者：王以诺 蔡施霞[2] 李连弟[2] 吕坤聚[1] WANG Yinuo;CAI Shixia;LI Liandi;L Kunju(Department of Respiratory Medicine,971 Hospital of the People’s Liberation Army of China,Qingdao 2660071,China)

机构地区：[1]中国人民解放军第九七一医院呼吸内科,山东青岛266071 [2]青岛大学附属医院重症医学科

出　　处：《青岛大学学报（医学版）》2023年第6期796-801,共6页Journal of Qingdao University(Medical Sciences)

基　　金：国家自然科学基金青年科学基金项目(81701879)。

摘　　要：目的探讨急性呼吸窘迫综合征(ARDS)时骨髓间充质干细胞(BMSC)旁分泌角质细胞生长因子(KGF)对脂多糖(LPS)损伤的肺泡上皮的修复作用,以及肺泡上皮细胞的晚期糖基化终末产物受体(RAGE)在BMSC旁分泌KGF修复LPS诱导的肺泡上皮损伤中的作用。方法建立LPS诱导的小鼠肺上皮细胞(MLE12)损伤模型以及体外BMSC与MLE12间接共培养模型。实验对象分为control、LPS、MSC-control、MSC-LPS、KGF及anti-RAGE组,培养1、7 d后,应用蛋白质印迹方法测定MLE12细胞中RAGE及闭锁小带蛋白-1(ZO-1)的含量,应用酶联免疫吸附试验方法测定细胞条件培养液中KGF及RAGE的含量。结果BMSC能增加LPS损伤肺上皮的ZO-1蛋白表达(F=172.40、146.80,P<0.01;t=14.91~67.40,P<0.05)。BMSC修复LPS损伤的肺上皮时条件培养液中KGF表达增加(F=45.51、2359.00,P<0.01;t=14.87~73.25,P<0.05);MLE12细胞的RAGE表达增加,差异有统计学意义(F=49.90~275.60,P<0.01;t=11.02~57.84,P<0.05)。结论BMSC对LPS诱导的MLE12细胞损伤有修复作用,BMSC修复LPS诱导的MLE12细胞损伤可能有旁分泌KGF的参与并通过调节RAGE表达实现。Objective To investigate the repair effect of keratinocyte growth factor(KGF)secreted by bone marrow mesenchymal stem cells(BMSC)through a paracrine mechanism on lipopolysaccharide(LPS)-induced alveolar epithelial damage in acute respiratory distress syndrome(ARDS),as well as the role of receptor for advanced glycation end products(RAGE)from alveolar epithelial cells on the repair of LPS-induced alveolar epithelial damage by KGF secreted by BMSC.Methods Mouse lung epithelial MLE12 cells were used to establish a model of LPS-induced damage,and an indirect co-culture model was established for BMSC and MLE12 in vitro.The subjects were divided into control group,LPS group,MSC-control group,MSC-LPS group,KGF group,and anti-RAGE group.After 1 and 7 days of culture,Western blot was used to measure the content of RAGE and zonula occluden-1(ZO-1)in MLE12 cells,and ELISA was used to measure the content of KGF and RAGE in the conditioned medium of cells.Results BMSC could increase the protein expression of ZO-1 in lung epithelium damaged by LPS(F=172.40,146.80,P<0.01;t=14.91-67.40,P<0.05).During the repair of lung epithelium damaged by LPS with BMSC,there were increases in the content of KGF in conditioned medium(F=45.51,2359.00,P<0.01;t=14.87-73.25,P<0.05)and the expression of RAGE in MLE12 cells(F=49.90-275.60,P<0.01;t=11.02-57.84,P<0.05).Conclusion BMSC can repair the damage of MLE12 cells induced by LPS,and KGF secreted by BMSC through a paracrine mechanism plays an important role in such repair and may help to achieve the repair effect by regulating the expression of RAGE.

关 键 词：间质干细胞 呼吸窘迫综合征 成纤维细胞生长因子10 高级糖化终产物受体 脂多糖类 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R563.8[医药卫生—基础医学]