环境与职业医学2025，Vol.42Issue(4)：482-489,8.DOI:10.11836/JEOM24405
过表达Ptpn2抑制SiO2介导的肺泡Ⅱ型上皮细胞炎症应答
Overexpression of Ptpn2 inhibits SiO2-mediated inflammatory response in alveolar type Ⅱ ep-ithelial cells
摘要
Abstract
[Background]Protein tyrosine phosphatase non-receptor type Ⅱ(PTPN2)is essential for the reg-ulation of inflammation and immunity,but the specific mechanism of action of Ptpn2 in silicosis is unknown. [Objective]To investigate the regulatory role of overexpression of Ptpn2 in SiO2-mediated in-flammatory response in alveolar type Ⅱ epithelial cells based on transcriptome sequencing. [Methods]This study was an in vitro study.A negative control group(vector transferred)and an overexpression of Ptpn2 group of mouse lung epithelial cell line MLE-12 cells were firstly constructed.Transcriptome sequencing was performed to detect differentially expressed genes(DEGs),differentially expressed mRNAs,and differentially expressed ncRNAs in the two groups of MLE-12 cells,and then the DEGs were analyzed by the Gene Ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG).Constructed MLE-12 cells and A549 cells were stimulated using SiO2 suspension,and divided into a negative control group(vector transferred),an overexpression of Ptpn2 group,a negative control+SiO2 group,and an overexpression of Ptpn2+SiO2 group,respectively.Protein expressions of tumor necrosis factor-α(TNF-α)and interleukin(IL)-17A,IL-2,IL-1β were detected by Western blot.Positive TNF-α expression was detected by immunofluorescence staining. [Results]The results of Western blot showed that the protein expression level of PTPN2 was up-regulated in the overexpressed Ptpn2 group compared with the negative control group(P<0.05).The volcano plot and clustering heat map showed that there were 1122 DEGs,3681 differentially expressed mRNAs,and 474 differentially expressed ncRNAs in the overexpressed Ptpn2 group compared with the negative control group.The results of GO enrichment showed that,in terms of the biological process,the DEGs were mainly related to the regulation of metal ion transport,extracellular matrix organization,and extracellular structural organization;in terms of cellular composition,the DEGs were mainly related to collagen-containing extracellular matrix,receptor complexes,and basement membranes;in terms of molecular function,the DEGs were mainly related to the extracellular matrix structural constituents,glycosaminoglycan bind-ing,and semaphorin receptor binding.The results of KEGG enrichment showed that the DEGs were closely related to cytokin-cytokine re-ceptor interaction,IL-17 signaling pathway,TNF signaling pathway,and chemokine signaling pathway.In MLE-12 and A549 cells,the results of Western blot showed that the protein expression levels of TNF-α,IL-17A,IL-2,and IL-1β were up-regulated in the negative control+SiO2 group compared with the negative control group(P<0.05),and the protein expression levels of TNF-α,IL-17A,IL-2,and IL-1β were down-regulated in the overexpression of Ptpn2+SiO2 group compared with the negative control+SiO2 group(P<0.05).The immunoflu-orescence staining showed that the expression of TNF-α was increased in the negative control+SiO2 group compared with the negative control group,and decreased in the overexpression of Ptpn2+SiO2 group compared with the negative control+SiO2 group. [Conclusion]Overexpression of Ptpn2 inhibits SiO2-mediated inflammatory response in MLE-12 cells and A549 cells.关键词
蛋白酪氨酸磷酸酶非受体Ⅱ型/矽肺纤维化/转录组测序/上皮细胞/炎症Key words
protein tyrosine phosphatase non-receptor type Ⅱ/silicosis fibrosis/transcriptome sequencing/epithelial cell/inflammation分类
预防医学引用本文复制引用
冯梦斐,魏懿,孙鑫茹,宫景硕,高学敏,徐洪,朱莹..过表达Ptpn2抑制SiO2介导的肺泡Ⅱ型上皮细胞炎症应答[J].环境与职业医学,2025,42(4):482-489,8.基金项目
国家自然科学基金项目(82204006) （82204006）
河北省自然科学基金项目(H2021209049) （H2021209049）
