杨少奇, 史晓妮, 成于思, 巢杰. 基于单细胞转录组测序探索FABP5在矽肺纤维化中II型肺泡上皮细胞的特异性表达[J]. 环境与职业医学, 2022, 39(10): 1089-1094. DOI: 10.11836/JEOM22126
引用本文: 杨少奇, 史晓妮, 成于思, 巢杰. 基于单细胞转录组测序探索FABP5在矽肺纤维化中II型肺泡上皮细胞的特异性表达[J]. 环境与职业医学, 2022, 39(10): 1089-1094. DOI: 10.11836/JEOM22126
YANG Shaoqi, SHI Xiaoni, CHENG Yusi, CHAO Jie. Specific expression of FABP5 in type II alveolar epithelial cells in silicosis pulmonary fibrosis based on single-cell transcriptome sequencing[J]. Journal of Environmental and Occupational Medicine, 2022, 39(10): 1089-1094. DOI: 10.11836/JEOM22126
Citation: YANG Shaoqi, SHI Xiaoni, CHENG Yusi, CHAO Jie. Specific expression of FABP5 in type II alveolar epithelial cells in silicosis pulmonary fibrosis based on single-cell transcriptome sequencing[J]. Journal of Environmental and Occupational Medicine, 2022, 39(10): 1089-1094. DOI: 10.11836/JEOM22126

基于单细胞转录组测序探索FABP5在矽肺纤维化中II型肺泡上皮细胞的特异性表达

Specific expression of FABP5 in type II alveolar epithelial cells in silicosis pulmonary fibrosis based on single-cell transcriptome sequencing

  • 摘要: 背景

    矽肺是由于长期吸入大量游离的二氧化硅(SiO2)颗粒所致,探讨其机制可以为矽肺纤维化的治疗提供新的方向。

    目的

    本研究着重探讨脂肪酸结合蛋白5(FABP5)在SiO2诱导的矽肺模型中的表达及发挥的作用。

    方法

    结合前期单细胞转录组测序结果,利用生物信息分析技术对小鼠肺泡上皮细胞进行分群并探索FABP5的表达模式;采用空间转录组学技术探索fabp5的分布模式。小鼠气管滴注SiO2建立矽肺纤维化模型,设置4个组别:生理盐水(normal saline, NS)7 d组、NS 56 d 组、SiO2 7 d组、SiO2 56 d组。SiO2处理小鼠肺上皮细胞系(MLE-12)建立矽肺体外模型。在动物整体水平,利用组织免疫荧光实验检测上皮细胞的标志物(E-Cad)和FABP5的蛋白水平;在体外水平,利用MLE-12验证细胞模型中fabp5 mRNA 表达变化和蛋白水平变化。

    结果

    单细胞转录组测序结果及空间转录组测序结果显示在小鼠矽肺病灶区域II型肺泡上皮细胞中fabp5 mRNA表达上调,伴随着组织免疫荧光蛋白水平升高,且与E-Cad存在共定位现象。同时SiO2刺激诱导MLE-12细胞中fabp5 mRNA表达升高至1.58倍和蛋白水平上升至2倍,差异均具有统计学意义(P<0.05)。

    结论

    在肺纤维化模型的肺泡上皮细胞中,FABP5的蛋白水平上升,提示FABP5可能参与上皮细胞在肺纤维化病理进程。

     

    Abstract: Background

    Silicosis is caused by long-term inhalation of large amounts of free silica (SiO2) particles, and exploring its mechanism can provide new directions for the treatment of silicosis fibrosis.

    Objective

    To investigate the expression and role of fatty acid binding protein 5 (FABP5) in a silica-induced silicosis model.

    Methods

    In combination with the results of single-cell transcriptome sequencing, the expression pattern of FABP5 in mouse alveolar epithelial cells was explored by bioinformatic analysis, and the distribution pattern of fabp5 was detected by spatial transcriptomics. An in vivo model of silicosis was established by intratracheal injection with SiO2 into mice and four groups were set up: normal saline (NS) 7 d group, NS 56 d group, SiO2 7 d group, and SiO2 56 d group. An in vitro model of silicosis was established in SiO2-treated mouse lung epithelial cell line (MLE-12). At the whole animal level, the marker of epithelial cells (E-Cad) and the protein level of FABP5 were detected by tissue immunofluorescence assay; in vitro, the changes of fabp5 mRNA expression and protein level in MLE-12.

    Results

    The results of single-cell transcriptome sequencing and spatial transcriptome sequencing showed that the mRNA expression of fabp5 was upregulated in type II alveolar epithelial cells in the focal area of silicosis in mice, accompanied by elevated tissue immunofluorescent protein levels, and there was co-localization of E-CAD. Meanwhile, SiO2 stimulation induced a 1.58-fold increase in fabp5 mRNA expression and a 2-fold increase in protein levels in MLE-12 cells, with significant differences (P<0.05).

    Conclusion

    The protein level of FABP5 is increased in alveolar epithelial cells in a pulmonary fibrosis model, suggesting that FABP5 may be involved in the pathological process of epithelial cells in pulmonary fibrosis.

     

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