炎症标志物C5a调控M2型巨噬细胞极化的机制及其临床应用价值  

Mechanism and clinical value of inflammatory marker C5a regulating the polarization of M2 macrophages

作　　者：李洁 应帅 胡雨琴 徐建 谢梦晓 LI Jie;YING Shuai;HU Yuqin;XU Jian;XIE Mengxiao(Department of Laboratory Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,Jiangsu;Department of Immunology,Nanjing Medical University School of Basic Medical Sciences,Nanjing 211166,Jiangsu,China)

机构地区：[1]南京医科大学第一附属医院检验学部,南京210029 [2]南京医科大学基础医学院免疫学系,南京211166

出　　处：《临床检验杂志》2025年第3期204-208,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(82101902);江苏省自然科学基金(BK20201079)。

摘　　要：目的 探讨炎症标志物C5a调控M2型巨噬细胞极化的作用机制及其在肺癌预后评估中的临床应用价值。方法 分别利用C5a刺激佛波酯(PMA)处理的人单核细胞白血病细胞系THP-10、6、12、24和48 h或0、1、2、3、6和12 h后,采用实时荧光定量PCR(qRT-PCR)测定M2型标志物CD163和CD206的mRNA表达水平;Western blot测定组蛋白乙酰基转移酶——通用控制非抑制蛋白5(GCN5)的表达水平;免疫共沉淀(co-IP)测定巨噬细胞关键转录因子GATA结合蛋白3(GATA3)乙酰化及其与GCN5、E1A结合蛋白p300(Ep300/p300)的结合能力;利用GCN5抑制剂丁内酯3预处理巨噬细胞后行C5a刺激,采用Western blot和co-IP技术分别测定CD163和CD206的表达水平、GATA3的乙酰化及其与GCN5的结合能力;通过KM plotter数据库分析肺癌组织中C5a受体1(C5aR1)与GCN5的表达水平在肺癌预后评估中的临床意义。结果 在PMA诱导贴壁的巨噬细胞中,C5a上调巨噬细胞中CD163和CD206的mRNA水平、GCN5的表达水平、GATA3的乙酰化以及两者的结合能力,但不影响GATA3与p300的结合能力;抑制GCN5活性不仅能显著降低GATA3的乙酰化及其与GCN5的结合能力,还能下调CD163和CD206的表达水平;高表达C5aR1或GCN5肺癌患者的总体生存率明显降低。结论 C5a通过促进GCN5乙酰化GATA3,促进M2型巨噬细胞极化,肺癌组织中C5aR1和GCN5的表达对患者的预后评估具有一定的临床应用价值。Objective To investigate the role and mechanism of inflammatory marker C5a in regulating the polarization of M2 macrophages as well as its clinical application value in the prognosis evaluation of lung cancer.Methods The phorbol 12-myristate 13-acetate(PMA)-pulsed human monocytic leukemia cell line THP-1 was stimulated with C5a for 0,6,12,24,and 48 h or 0,1,2,3,6,and 12 h,and then the expression levels of M2 markers CD163 and CD206 mRNA were determined by real-time fluorescence quantitative PCR(qRT-PCR).The expression level of general control non-repressed protein 5(GCN5)with histone acetyltransferase(HAT)activity was detected by Western blot.Co-immunoprecipitation(co-IP)was used to determine the acetylation of GATA binding protein 3(GATA3),a key transcription factor for macrophages,and its binding ability to GCN5 and E1A binding protein p300(Ep300/p300).After the macrophages pre-treated with GCN5 inhibitor butyrolactone 3(MB-3)were stimulated with C5a,the expression levels of CD163 and CD206,acetylation of GATA3,and its binding ability to GCN5 were determined by Western blot and co-IP.The clinical significance of the expression of C5a receptor 1(C5aR1)and GCN5 in lung cancer tissues in the prognosis of lung cancer patients was analyzed using the KM plotter database.Results C5a could significantly increase the expression levels of CD163 and CD206 mRNA,expression of GCN5,acetylation of GATA3,and its binding ability to GCN5 in PMA-induced adherent macrophages,but did not affect the binding of GATA3 to p300.Inhibiting the activity of GCN5 not only significantly reduced the acetylation of GATA3 and its binding ability to GCN5,but also down-regulated the expressions of CD163 and CD206.The overall survival rate of lung cancer patients with high expression of C5aR1 or GCN5 was significantly reduced.Conclusion C5a promotes the polarization of M2 macrophages by inducing GCN5 to acetylate GATA3.The expressions of C5aR1 and GCN5 in lung cancer tissues may have certain clinical application value for the prognosis of the pati

关 键 词：肺癌 巨噬细胞 C5A 乙酰化 

分 类 号：R446.6[医药卫生—诊断学] R392.11[医药卫生—临床医学]