靶向性荧光示踪间充质干细胞治疗急性肺损伤

发布时间：2018-02-24 05:22

本文关键词： 荧光探针示踪间充质干细胞急性肺损伤聚集　出处：《中山大学学报(医学科学版)》2017年03期 　论文类型：期刊论文

【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。
[Abstract]:[objective] to establish a mouse model of acute lung injury (Ali) and to treat acute lung injury with the tracer of mesenchymal stem cells (MSCs). [methods] Anti-GD2, a monoclonal antibody targeting MSC surface ganglioside GD2, was used as vector. A novel fluorescent molecular probe, Anti-GD2-Cy7, was obtained by covalent binding with Cy7. The synthesized fluorescent probe Anti-GD2-Cy7 was labeled on MSC by antigen-antibody reaction. 84 male Balb/c mice were used in this study. 48 of them were randomly divided into three groups: sham operation group (ALI group) and NS ALI group (n = 16). Lung histomorphology and pathological score were observed at 24 h and 48 h after the establishment of ALI model. Lung wet / dry weight / W / D ratio and pulmonary microvascular permeability were measured. The remaining 36 male Balb/c mice were randomly divided into normal group (n = 12), sham-operated group (n = 12) and MSC ALI group (n = 12). The labeled MSC-GD2-Cy7 was injected via tail vein into sham operation group and MSC ALI group. The anaesthetized mice were anaesthetized at 30 min and 1 d 3 d after caudal vein injection respectively. [results] the model was made up to the level of ALI, both 24 h after treatment and 48 h after treatment. The average fluorescence intensity of lung tissue in the MSC ALI group was significantly higher than that in the sham operation group [3.37 卤0.02) 脳 10 ~ (-4) VSD 2.05 卤0.05 卤0.04) 脳 10 ~ (10) -4scaled countsrsr (35.54 卤0.47) 脳 10 ~ (-4) VS ~ (25.29 卤1.48) 脳 10 ~ (-4) cases / r = 11.17 卤0.75) 脳 10 ~ (10) ~ (-0. 75) 脳 10 ~ (-4) V ~ (-1) P = 6.09 卤0.62) 脳 10 ~ (10) ~ (-0. 14) 卤0. 10 卤0. 14 卤0. 14 卤0. 14 卤0. 00 卤0. 14 卤0. 05 卤0. 14%. [P < 0. 05] [P < 0. 05] [P < 0. 05] [P < 0. 05] [P < 0. 05] [P < 0. 05]. The stem cells migrated to the normal and injured lungs 30 min after injection. On the first day, the MSC was accumulated in the injured lung and lasted until the 7th day, which indicated that MSC had the effect of aggregation at the injured site.
【作者单位】：中山大学附属第三医院外科ICU;
【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)
【分类号】：R563.8

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