摘要
【目的】通过构建小鼠急性肺损伤(ALI)模型,示踪间充质干细胞(MSC)治疗急性肺损伤。【方法】以MSC表面神经节苷脂(GD2)特异性抗原靶向的单克隆抗体Anti-GD2为载体,与荧光菁染料(Cy7)共价结合,得到新型荧光分子探针Anti-GD2-Cy7。将合成的荧光分子探针Anti-GD2-Cy7通过抗原抗体反应标记在MSC上。本研究取84只Balb/c雄性小鼠,随机抽取其中48只平均分成三组:假手术组(n=16)、MSC+ALI组(n=16)和NS+ALI组(n=16)。建立ALI模型后,在24 h、48 h时间点进行肺组织形态学观察和病理评分、检测肺湿/干质量(W/D)比值和肺微血管通透性改变。剩余36只Balb/c雄性小鼠随机分为正常组(n=12)、假手术组(n=12)和MSC+ALI组(n=12),将标记上的MSC-GD2-Cy7通过尾静脉注射入假手术组和MSC+ALI组小鼠。分别在尾静脉注射后的30 min及1 d、3 d、7 d各时间点麻醉解剖小鼠后;取其肺在小动物荧光成像仪下进行检测。【结果】模型制备达到ALI水平,无论是治疗后的24 h还是48 h,MSC+ALI组在肺组织的病理及评分、肺组织的水肿程度和肺血管的通透性明显比NS+ALI组轻。荧光检测显示MSC+ALI组在各时间点肺部平均荧光强度显著高于假手术组[(3.37±0.02)×10^(-4)vs(2.05±0.04)×10^(-4)scaled counts/s;(35.54±0.47)×10^(-4)vs(25.29±1.48)×10^(-4)scaled counts/s;(11.17±0.75)×10^(-4)vs(6.09±0.62)×10^(-4)scaled counts/s;(3.10±0.14)vs(0.00±0.00)×10^(-4)scaled counts/s;且P均小于0.05)]。【结论】间充质干细胞在注射后的30 min即可迁移到正常肺和损伤肺中,且在第1天大量的聚集在损伤肺中并持续到第7天;这表明MSC具有在损伤部位聚集的作用。
[ Objective ] To investigate the mesenchymal stem cells (MSC) in treating acute lung injury (ALl) via ALI mouse model. [Methods] By monoclonal antibody Anti-GD2 of specific antigen ganglioside (GD2) only expressed on MSC as a carrier, new fluorescent molecule probe were synthesized through covalently coupling Anti-GD2 and fluorescent group CyDye mono-reactire NHS Esters (CyT). Synthetic Anti-GD2-Cy7 and MSC were labeled by the specific binding of antigen and antibody in vitro. Total 84 balb/c male mice were selected and randomly selected 48 mice were divided into three groups: sham group (n = 16), MSC+ ALl group(n = 16), NS + ALl group(n = 16). The lung histopathology and scores, lung W/D ratio and permeability of lung microvasculature were examined at 24 h, 48 h after ALl mouse model. Other 36 mice were randomly divided into three groups : normal group (n = 12) , sham group(n = 12) , MSC + ALl group(n = 12). Labeled MSC-GD2-Cy7 were transplanted into MSC+ALI group and sham group mice via tail vein injection. At 30 min, 1 d, 3 d, and 7 d post-MSC-GD2-Cy7 injection, the mice were sacrificed after anesthesia and then the lung was removed. Excised lung was detected on small animal fluorescent imager. [Results ] Contrast to NS+ ALI group, the lung histopathology and scores, lung W/I) ratio and permeability of lung microvasculature of MSC +ALI group were more greatly improved at both 24 h and 48 h. Fluorescent results showed that the signal intensity in the lung of MSC + ALI group was significantly higher than that of sham group at each time point [(3.37 ± 0.02)× 10^-4 vs (2.05 ± 0.04) × 10^-4 scaled counts/s; (35.54 ± 0.47)× 10 ^-4vs (25.29 ± 1.48) × 10^-4 scaled counts/s; (11.17 ± 0.75)×10^-4 vs (6.09 ± 0.62)× 10^-4 scaled counts/s; (3.10 ± 0.14) vs (0.00 ± 0.00) × 10^-4 scaled counts/s; all P 〈 0.05 ]. [ Conclusion ] Our study showed that a proportion of cells migrated into normal and injured lungs 30 rain after cell transplantation, and the cells started to recruit and largely gather in injured lungs at day 1 and persisted to day 7, these results suggest that MSC possess the ability to home into injured tissues.
出处
《中山大学学报(医学科学版)》
CAS
CSCD
北大核心
2017年第3期332-340,共9页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省科技计划项目(2014A020211010)
广州市天河区科技计划项目(2013kw028)
关键词
荧光探针
示踪
间充质干细胞
急性肺损伤
聚集
fluorescent probe
visualization
mesenchymal stem cells
acute lung injury
gather