骨髓间充质干细胞对脂多糖诱导的急性肺损伤小鼠的早期治疗作用(英文)

Therapeutic effect of intravenous bone marrow-derived mesenchymal stem cell transplantation on early-stage LPS-induced acute lung injury in mice

目的观察经尾静脉输注的骨髓间充质干细胞(MSCs)对脂多糖(LPS)诱导的急性肺损伤小鼠的早期治疗作用。方法采用密度梯度单个核细胞分离培养法分离纯化骨髓间充质干细胞,传至第4代时观察细胞形态、流式细胞仪检测细胞表面标志,定向诱导检测分化能力后备用。36只小鼠随机分为对照组,PBS治疗组,MSC治疗组。对照组气管内滴注生理盐水50μl,PBS和MSC治疗组气管内滴注LPS制备急性肺损伤模型。造模后1 h后经尾静脉输注MSC5×106(MSCs治疗组)或PBS 100μ(lPBS治疗组或对照组)。24 h后处死小鼠,留取标本检测肺组织病理形态学、肺组织湿干重比(W/D)、支气管肺泡灌洗液(BALF)中中性粒细胞数、蛋白含量、细胞因子水平和肺组织中的MPO表达量。结果与对照组比较,PBS治疗组肺组织病理损伤严重,BALF中中性粒细胞数、蛋白含量、TNF-α、IL-6和IL-10水平、肺组织中的MPO含量及肺组织湿干重比均显著升高。与PBS治疗组比较,MSCs治疗组肺组织病理损伤程度减轻,BALF中中性粒细胞数、蛋白含量、TNF-α、IL-6水平、肺组织中的MPO含量及肺组织湿干重比均显著降低,但仍高于对照组水平,IL-10水平显著高于PBS治疗组和对照组。结论 MSCs移植在LPS诱导的急性肺损伤早期阶段可有效改善肺组织病理损伤,减轻肺部炎症反应,降低肺水肿程度,这种治疗作用不依赖于MSCs移植后在肺内的定植数量。

Objective To investigate the effect of intravenous bone marrow-derived mesenchymal stem cell（MSC） transplantation for early intervention of lipopolysaccharide（LPS）-induced acute lung injury（ALI） in mice.Methods Thirty-six mice were randomized into control group,PBS-treated ALI group,and MSC-treated ALI group.In the latter two groups,mouse models of ALI were established by intranasal instillation of LPS,and 1 h later,the 4th passage of MSCs isolated from the bone marrow of mice or PBS were administered via the tail vein.The histological findings,lung wet/dry（W/D） weight ratio,neutrophil count and protein and cytokine contents in the bronchoalveolar lavage fluid（BALF）,and myeloperoxidase（MPO） level in the lung tissue were analyzed at 24 h after MSC administration.Engraftment of MSCs in the recipient lung was determined by fluorescent PKH26 staining and flow cytometry.Results Compared with the control group,PBS-treated ALI group showed significantly higher protein levels,tumor necrosis factor-α（TNF-α）,interleukin-6（IL-6） and neutrophil count in the BALF and MPO content in the lung tissue,with also severe damage of lung histology.MSCs administration significantly reduced the lung W/D weight ratio,the levels of protein,TNF-α,IL-6 and neutrophil count in the BALF and MPO content in the lung tissue,and obviously lessened the lung injury 24 h after the transplantation.MSC administration also significantly increased the level of IL-10 in the BALF.Conclusion Intravenous MSC transplantation can effectively improve the lung histology,attenuate the inflammatory response,reduce pulmonary edema in the early stage of LPS-induced ALI in mice,and such effects are independent of MSC engraftment in the lungs.