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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南京医科大学附属常州第二人民医院

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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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The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University

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Jiangsu province social development project (BE2020651 to Q.Z.),The Fifth "333" High-level Talent Project of Jiangsu Province (BRA2020015),Changzhou High-Level Medical Talents Training Project (2022CZLJ013 to Q.Z.),Research Project of Changzhou Medical Center of Nanjing Medical University(CMCB202214)

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    摘要:

    目的:研究慢性阻塞性肺疾病(COPD)患者外周血α 2巨球蛋白(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分析被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相关性。ELISA用于检测血浆中A2M的浓度。实时荧光定量PCR法检测细胞和外周血中A2M mRNA的表达水平。蛋白免疫印迹试验检测M2型巨噬细胞表面标记物精氨酸酶(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的诊断敏感性与特异性。 结果:对GSE38974数据集的差异表达基因(DEGs)进行生信分析,发现A2M在COPD肺组织中表达下降,且与COPD肺组织免疫细胞浸润相关。RT-qPCR和ELISA结果显示A2M水平在COPD患者外周血中下调,外周血中A2M蛋白水平与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。敲低A2M后,M2巨噬细胞的表面标记物Arg-1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: The aim of this study is to investigate the correlation between α 2 macroglobulin (A2M) and immune cell infiltration in peripheral blood of patients with chronic obstructive pulmonary disease (COPD). Methods: The GSE38974 dataset was comprehensively analyzed. GO enrichment, KEGG analysis, and GSVA analysis were used to explore potential functions and pathways. CIBERSORT was used to assess tissue infiltrating immune cells. A total of 25 patients with stable COPD and 26 healthy controls were enrolled in this study. The correlation between peripheral blood A2M level and immune cell count was analyzed. ELISA was used to measure the concentration of A2M in plasma. The expression of A2M mRNA in cells and peripheral blood was detected by real-time fluorescence quantitative PCR. The expression of arginase 1 (Arg-1), a surface marker of M2 macrophages, was detected by Western blot. 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 (DEGs) in GSE38974 dataset showed that the expression of A2M was decreased in COPD lung tissue, and was related to immune cell infiltration in COPD lung tissue. RT-qPCR and ELISA results showed that the level of A2M was down-regulated in peripheral blood of COPD patients, and the level of A2M protein in peripheral blood was correlated with lymphocytes and monocytes in COPD patients. ROC curve 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. Knockdown of A2M in M2 macrophages reduced the expression of Arg-1, a surface marker of M2 macrophages. Conclusions: The expression of A2M in lung tissue and peripheral blood of COPD patients is decreased, which is closely related to the immune cell count and immune cell infiltration in COPD patients. A2M may play an important role in the polarization of macrophages to M2 type.

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  • 收稿日期:2023-06-13
  • 最后修改日期:2023-11-22
  • 录用日期:2023-12-13
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