骨髓间充质干细胞对小鼠急性肺损伤治疗作用的研究

目的探讨输入外源性骨髓间充质干细胞(MSCs)对脂多糖(LPS)诱导的小鼠急性肺损伤(AL)I的治疗作用.方法实验随机分为3组(n=10):对照组、ALI组和MSCs治疗组.健康小鼠LPS气管滴入方法建立ALI模型,尾静脉输入全骨髓培养法分离纯化的MSCs.流式细胞分析鉴定MSCs.HE染色组织切片观察输入MSCs前后肺组织病理形态学改变,W/D比值和髓过氧化物酶(MPO)活性评价肺水肿和炎症反应程度.ELISA检测肺组织匀浆中肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)的含量.结果流式细胞分析鉴定:培养MSCs表面标记CD105为阳性,CD34为阴性.肺组织病理学显示:ALI组小鼠肺组织呈现典型的炎症病理变化,包括肺泡充血、出血、水肿,肺泡腔和血管壁中性粒细胞浸润,肺泡壁增厚等肺损伤性病变;MSCs治疗组小鼠肺组织损伤程度明显减轻.肺W/D比值、肺组织MPO活性及TNF-α和IL-6含量:ALI组小鼠肺W/D比值、肺组织MPO活性及TNF-α和IL-6含量高于对照组(P<0.05);MSCs治疗组肺W/D比值、肺组织MPO活性及TNF-α和IL-6含量低于ALI组(P<0.05).结论输入MSCs能够减轻脂多糖致急性肺损伤程度,其作用可能与降低肺W/D比值和肺组织MPO活性、IL-6及TNF-a含量有关.

The Treatment Effect of Marrow-derived Mesenchymal Stem Cells on LPS -induced Acute Lung Injury in Mice

Objective To study treatment effects of extrinsic mesenchymal stem cells(MSCs)on mice with acute lung injury(ALI)induced by lipopolysaccharide(LPS).Methods Thirthy mice was divided randomly into 3 groups(n=10):control group,ALI group and MSCs therapy group.Healthy mice were given lipopolysaccharide(LPS)intratracheally to cause acute lung injury,MSCs from mouce bone marrow were isolated,cultured and amplified in vitro,and the MSCs were given to mice via caudal vein.MSCs marker were identificated by flow cytometry.Pathomorphological changes of mouce lung were observed under light microscope after Hematoxylin-Eosin staining.The lung wet to dry weight ratio and the lung tissue MPO activity were determinated to assessed lung edema and inflammatory degree.The concertrations of IL-6 and TNF-α in lung tissue homogenate were measured by ELISA.Results Fow cytometer analysis showed that the cultured MSCs displayed a positive CD105 and a negative CD34.Histopathological evalution showed that typical pathological inflammation lesions in the lung were observed in ALI group,including alveolar congestion,hemorrhage,edema,infiltration of neutrophils in the airspace or vessel wall,thickness of the alveolar wall;pathological changes were relieved obviously in MSCs therapy group.The changes of lung W/D weight ratio and the lung tissue MPO activity,IL-6 and TNF-α content of lung tissues homogenate in mice in ALI group were all higher than those in control group(P<0.05);the lung W/D weight ratio,IL-6 and TNF-α content in MSCs groups were all lower than those in ALI group(P<<0.05).Conclusion MSCs can alleviate injury degree of ALI induced by LPS in mice,the mechanism of action may correlate with decreasing the lung W/D weight ratio and MPO activity,IL-6 and TNF-α content in lung tissue.