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

作　　者：杨少奇 史晓妮 成于思 巢杰 YANG Shaoqi;SHI Xiaoni;CHENG Yusi;CHAO Jie(Key Laboratory of Environmental Medicine Engineering,Ministry of Education/School of Public Health,School of Medicine,Southeast University,Nanjing,Jiangsu 210000,China;Department of Physiology,School of Medicine,Southeast University,Nanjing,Jiangsu 210000,China)

机构地区：[1]东南大学公共卫生学院/环境医学工程教育部重点实验室,江苏南京210000 [2]东南大学医学院生理学系,江苏南京210000

出　　处：《环境与职业医学》2022年第10期1089-1094,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目:(81972987)。

摘　　要：[背景]矽肺是由于长期吸入大量游离的二氧化硅(SiO_(2))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SiO_(2)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SiO_(2)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、SiO_(2)7 d组、SiO_(2)56 d组。SiO_(2)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SiO_(2)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。[Background]Silicosis is caused by long-term inhalation of large amounts of free silica(SiO_(2))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 SiO_(2)into mice and four groups were set up:normal saline(NS)7 d group,NS 56 d group,SiO_(2)7 d group,and SiO_(2)56 d group.An in vitro model of silicosis was established in SiO_(2)-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,SiO_(2)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.

关 键 词：脂肪酸结合蛋白5 矽肺 单细胞转录组测序 空间转录组测序 

分 类 号：R114[医药卫生—卫生毒理学]