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陈佳,黄燕霞,冯玉丽,刘贝珠.肺祖细胞抑制博来霉素诱导的小鼠肺纤维化[J].中国医药科学,2024,14(5):25-29        基金项目:广东省广州市番禺区科技计划项目(2020-Z04-100)
肺祖细胞抑制博来霉素诱导的小鼠肺纤维化
Inhibition of lung progenitor cells on mice pulmonary fibrosis induced by bleomycin
  
DOI:
中文关键词:  胚胎干细胞;肺祖细胞;肺纤维化;免疫缺陷小鼠
英文关键词:Embryonic stem cells; Lung progenitor cells; Pulmonary fibrosis; Immunodeficient mice
作者单位
陈佳,黄燕霞,冯玉丽,刘贝珠 广州市番禺区何贤纪念医院,广东广州 511400 
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中文摘要:
      [摘要]目的 研究肺祖细胞在小鼠肺内定植的情况及其对小鼠肺纤维化的影响。方法 人胚胎干细胞体外分化为肺祖细胞,经免疫荧光和细胞流式术鉴定后,移植至正常免疫缺陷小鼠肺内,观察其定植的时间以及是否在体内发生分化为成熟肺上皮细胞。免疫缺陷小鼠经气管灌注博来霉素(2 μg/10 g)建立肺纤维化模型,并将肺祖细胞移植至肺纤维化的小鼠肺内,治疗后取肺组织进行切片做苏木素-伊红染色和Masson染色,同时检测肺内低α-平滑肌肌动蛋白(α-SMA)、细胞外基质蛋白-1(ECM-1)和羟脯氨酸表达的变化,以及检测凋亡的情况。结果 免疫荧光和细胞流式术显示,定型内胚层细胞阶段标志物叉头框蛋白A2和趋化因子受体4的表达率能达到90%,肺祖细胞标志物NKX2.1显示肺祖细胞的分化效率为70%;在移植后4周,在免疫缺陷小鼠肺内找到定植的肺祖细胞,但未发现肺祖细胞表达表面活性物质C;灌注肺祖细胞后,各组间反映肺纤维化程度的Aschcroft score、α-SMA、ECM-1比较,差异有统计学意义(P < 0.01);各组间反映肺上皮功能的表面活性物质C荧光面积比较,差异有统计学意义(P < 0.01);各组间反映胶原水平高低的羟脯氨酸浓度比较,差异有统计学意义(P < 0.01);反映细胞凋亡的原位末端转移酶标记技术染色实验,各组间荧光面积比较,差异有统计学意义(P < 0.01)。结论 肺祖细胞可以在免疫缺陷小鼠肺内定植,不能分化为成熟的肺上皮细胞,但能抑制肺纤维化的进展。
英文摘要:
      [Abstract] Objective To investigate the colonization of lung progenitor cells in the lungs of mice and their effects on mice pulmonary fibrosis. Methods Human embryonic stem cells were differentiated into lung progenitor cells in vitro, identified by immunofluorescence and cell flow cytometry, and transplanted into the lungs of normal immunodeficient mice. The time of colonization and the ability to differentiate into mature lung epithelial cells in vivo were observed. Immunodeficient mice were perfused with bleomycin (2 μg/10 g) through the air duct to establish a pulmonary fibrosis model, and lung progenitor cells were transplanted into the lungs of the mice with fibrosis. After treatment, lung tissue was sliced for hematoxylin-eosin staining and Masson staining, and changes of low α-smooth muscle actin (α-SMA), extracellular matrix protein 1 (ECM-1), and hydroxyproline expressions in lung and apoptosis were also tested. Results Immunofluorescence and cell flow cytometry showed that the expression rate of the definitive endoderm cell stage markers, forkhead box protein A2 and chemokine receptor 4, could reach over 90%. The lung progenitor cell marker NKX2.1 showed a differentiation efficiency of 70% for lung progenitor cells. At 4 weeks after transplantation, lung progenitor cells that were colonized in the lungs of immunodeficient mice were found, but no expression of surfactant protein C was found in lung progenitor cells. After perfusion of lung progenitor cells, there were statistical differences in the Ashcroft score, α-SMA and ECM-1 that reflect the degree of pulmonary fibrosis between the groups (P < 0.01). There was a difference in the fluorescence area of surfactant protein C that reflected the function of pulmonary epithelium between the groups (P < 0.01). There was a difference in the concentration of hydroxyproline that reflected the level of collagen between the groups (P < 0.01). There was a difference in the fluorescence areas between the groups in the in situ terminal deoxynucleotidyl transferase labeling technique staining experiment (P < 0.01). Conclusion Lung progenitor cells can be colonized in the lungs of immunodeficient mice and cannot differentiate into mature lung epithelial cells, but can inhibit the progression of pulmonary fibrosis.
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