A2M在慢性阻塞性肺疾病中的表达及其与免疫细胞浸润的相关性
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江苏省社会发展项目(BE2020651);江苏省第五期“333”高层次人才项目(BRA2020015);常州市高层次医学人才项目(2022CZLJ013);南京医科大学常州医学中心科研项目(CMCB202214)


The expression of A2M in chronic obstructive pulmonary disease and its correlation with immune cell infiltration
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    目的:研究慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者外周血α2巨球蛋白(alpha 2 macro- globulin,A2M)与免疫细胞浸润的相关性。方法:对GSE38974数据集进行综合分析。GO富集、KEGG分析和GSVA分析用于探索潜在的功能和通路。CIBERSORT用于评估组织浸润性免疫细胞。收集25例稳定期COPD患者和26例健康对照,分析外周血A2M水平与免疫细胞计数的相关性。ELISA检测血浆中A2M的浓度。RT-qPCR检测细胞和外周血中A2M mRNA的表达水平。Western blot法检测M2型巨噬细胞表面标志物精氨酸酶(arginase-1,Arg-1)的表达水平。采用Pearson相关分析进行相关性分析。ROC曲线判断A2M的诊断灵敏度与特异度。结果:对GSE38974数据集的差异表达基因进行生物信息学分析发现,A2M在COPD患者肺组织中表达下降,且与COPD患者肺组织的免疫细胞浸润相关。RT-qPCR和ELISA结果显示,A2M水平在COPD患者外周血中下调,与COPD患者淋巴细胞、单核细胞具有相关性。ROC曲线提示A2M具有诊断COPD的价值。进一步的通路分析提示A2M可能与巨噬细胞等调节免疫途径相关。在M2巨噬细胞中敲低A2M后Arg-1的表达降低。结论:A2M 在 COPD患者肺组织与外周血中表达降低,与COPD患者免疫细胞计数、免疫细胞浸润等密切相关,A2M可能在巨噬细胞向 M2型极化的过程中发挥了重要作用。

    Abstract:

    Objective:To investigate the correlation between peripheral blood alpha 2 macroglobulin(A2M)and immune cell infiltration in patients with chronic obstructive pulmonary disease(COPD). Methods:Comprehensive analysis of the GSE38974 dataset was performed. GO enrichment,KEGG analysis,and GSVA analysis were used to explore potential functions and pathways. CIBERSORT was used to assess tissue- infiltrating immune cells. Peripheral blood A2M levels and immune cell counts were analyzed in 25 stable COPD patients and 26 healthy controls. ELISA was used to detect the concentration of A2M in plasma. RT-qPCR was used to measure A2M mRNA expression levels in cells and peripheral blood. Western blot was used to measure the expression levels of the M2 macrophage surface marker arginase-1(Arg-1). Pearson correlation analysis was used for correlation analysis. ROC curve was used to determine the diagnostic sensitivity and specificity of A2M. Results:Bioinformatics analysis of differentially expressed genes in GSE38974 dataset revealed that A2M expression was decreased in lung tissue of COPD patients and was correlated with immune cell infiltration in lung tissues of COPD patient. RT-qPCR and ELISA results showed that A2M levels were down-regulated in peripheral blood of COPD patients,and were correlated with lymphocytes and monocytes in COPD patients. ROC curve analysis showed that A2M had a diagnostic value for COPD. Further pathway analysis suggested that A2M might be associated with macrophages and other regulatory immune pathways. Knocking down of A2M in M2 macrophages resulted in decreased expression of Arg-1. Conclusion:The expression of A2M is decreased in lung tissue and peripheral blood of COPD patients and is closely related to the immune cell counts and immune cell infiltration in COPD patients. A2M may play an important role in the polarization of macrophages to the M2 phenotype.

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肖心儒,范亮,施宇佳,张倩. A2M在慢性阻塞性肺疾病中的表达及其与免疫细胞浸润的相关性[J].南京医科大学学报(自然科学版),2024,(1):1-11,59

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  • 收稿日期:2023-06-13
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  • 在线发布日期: 2024-01-12
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