上海交通大学学报（医学版）

›› 2009, Vol. 29 ›› Issue (10): 1157-.

• 论著（基础研究） • 上一篇    下一篇

不同剂量BMSC尾静脉输注对小鼠肺组织纤维化的影响

徐佳波1, 李燕芹1, 李 莉2, 刘 斌1, 熊剑飞1, 叶 青1   

  1. 1. 上海交通大学 医学院仁济医院呼吸科, 上海 200127；2. 同济大学 上海肺科医院呼吸科, 上海 200433
  • 出版日期:2009-10-25 发布日期:2009-10-26
  • 通讯作者: 李燕芹, 电子信箱: drliyanqin@126.com。
  • 作者简介:徐佳波（1977—）, 男, 主治医师, 硕士；电子信箱: shonexu@yahoo.com.cn。
  • 基金资助:

    上海市科委自然基金（064119630）

Effects of different dosages of BMSC on lung fibrosis in mice

XU Jia-bo1, LI Yan-qin1, LI Li2, LIU Bin1, XIONG Jian-fei1, YE Qing1   

  1. 1. Department of Respiratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China；2. Department of Respiratory Medicine, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, China
  • Online:2009-10-25 Published:2009-10-26
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 064119630

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摘要：

目的 研究经尾静脉输注不同剂量的骨髓间充质干细胞（BMSC）对小鼠肺纤维化的影响。方法 体外培养雄性转红色荧光蛋白（RFP）基因FVB小鼠BMSC。将24只FVB雌性受体小鼠随机分为正常组、模型组、BMSC治疗1组（BMSC 1组）和BMSC治疗2组（BMSC 2组）（n=6）。模型组、BMSC 1组和BMSC 2组小鼠分别经气管内注射博莱霉素制备肺纤维化模型。建模24 h后，BMSC 1组和BMSCs 2组小鼠分别经尾静脉注射BMSC 1×106/只和2×106/只。于建模后第21天处死所有小鼠，制备小鼠肺组织标本。光学显微镜观察病理学变化；激光共聚焦显微镜和免疫组织化学法进行RFP(+)BMSC分布观察和RFP定量分析；免疫组织化学法检测肺表面活性物质蛋白A(SP-A)表达；碱水解法测定羟脯氨酸（Hyp）含量；Real-time PCR检测转化生长因子-β（TGF-β）和血小板衍生生长因子（PDGF）mRNA表达。结果 小鼠肺组织纤维化程度，BMSC 1组较模型组显著减轻，而BMSC 2组较模型组明显加重。BMSC 2组在肺间质纤维化活跃区域可见大量RFP(+)BMSC且其形态类似成纤维细胞。SP-A表达，正常组＞BMSC 1组＞BMSC 2组和模型组（均P<0.05），而BMSC 2组与模型组比较差异无统计学意义（P＞0.05）。Hyp含量，正常组＜BMSC 1组＜模型组＜BMSC 2组（P<0.01，P<0.05）；TGF-β和PDGF mRNA表达，BMSC 2组＞BMSC 1组＞模型组＞正常对照组（均P<0.05）。结论 适量的BMSC尾静脉输注能有效减轻博来霉素诱导的小鼠肺纤维化，而过量的BMSC则可能使其病变加重。

关键词: 骨髓间充质干细胞, 肺纤维化, 量效关系, 红色荧光蛋白, 肺表面活性物质蛋白, 羟脯氨酸, 转化生长因子-β,
血小板衍生生长因子

Abstract:

Objective To investigate the effects of different dosages of bone marrow mesenchymal stromal cells (BMSC) on lung fibrosis. Methods BMSCs with red fluorescence protein (RFP) from male FVB mice were cultured in vitro. Twenty-four female wild type FVB mice were randomly divided into four groups: normal group, model group, BMSC 1 group and BMSC 2 group (n=6). Mouse pulmonary fibrosis models were induced by bleomycin via single intratracheal perfusion. Twenty-four h after model establishment, mice in BMSC 1 group and BMSC 2 group were injected with 1×106 BMSCs and 2×106 BMSCs, respectively through vena caudalis for each mouse. All the animals were sacrificed 21 d after model estalishment, and mouse lung tissue samples were obtained. The pathological changes were observed by light microscopy, the hydroxyproline (Hyp) contents were measured by alkaline hydrolysis assay, the distribution of RFP(+) BMSCs and quantitation of RFP were analysed by laser scanning confocal microscopy and immunohistochemistry, the expression of surfactant protein A (SP-A) was detected by immunohistochemistry, and the expression of transforming growth factor-β (TGF-β) and platelet-derived growth factor (PDGF) mRNA was detected by Real-time PCR. Results Compared with model group, the pulmonary fibrosis in BMSC 1 group was significantly alleviated, and that of BMSC 2 group became much more severe. A large number of RFP(+) BMSCs were found in fibrosis area of BMSC 2 group, which exhibited morphology similar to fibroblasts. As far as the expression of SP-A was concerned, normal group was higher than BMSC 1 group, BMSC 1 group was higher than BMSC 2 group and model group (P<0.05), while there was no significant difference between BMSC 2 group and model group (P＞0.05). Normal group, BMSC 1 group, model group and BMSC 2 group fell in the increase order by Hyp contents (P<0.01, P<0.05), and BMSC 2 group, BMSC 1 group, model group and normal group fell in the decrease order by expression of TGF-β and PDGF mRNA (P <0.05）. Conclusion Proper dose of BMSC has a favourable effect on bleomycin-induced lung fibrosis, while excessive dose of BMSC can aggravate the fibrosis.

Key words: bone marrow mesenchymal stromal cell, lung fibrosis, dose-effective relationship, red fluorescence protein, surfactant protein, hydroxyproline, transforming growth factor-β, platelet-derived growth factor

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