MMP-7过表达调控巨噬细胞Gal-3分泌影响其极化和免疫应答  

Over-expression of MMP-7 regulates the secretion of Gal-3 in macrophages and influences their polarization and immune responses

作　　者：冯莹[1] 张妍 雷杰 刘娟[1] 方勇[1] 贺立群[1] FENG Ying;ZHANG Yan;LEI Jie;LIU Juan;FANG Yong;HE Li-qun(Department of Cardiology,Wuhan No.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院心内科,武汉430022

出　　处：《现代免疫学》2024年第5期375-381,共7页Current Immunology

基　　金：武汉市卫健委青年基金(WX21Q11)。

摘　　要：为研究基质金属蛋白酶7(matrix metalloproteinase 7,MMP-7)过表达对巨噬细胞极化、半乳糖凝集素3(galectin 3,Gal-3)表达和炎性因子分泌的影响,诱导THP-1细胞成巨噬细胞,转染MMP-7过表达和空载质粒,将实验细胞分为对照组、MMP-7过表达组、MMP-7 NC组、M1型诱导组和M2型诱导组。qPCR法检测Gal-3、MMP-7 mRNA表达水平;流式细胞术双标法检测F4/80^(+)、CD86^(+)、CD206^(+)巨噬细胞百分比;ELISA检测细胞上清液中炎性因子IL-1α、IL-6及TGF-β含量。使用Gal-3抑制剂处理M1型和M2型诱导分化巨噬细胞,qPCR法检测细胞中Gal-3 mRNA表达变化,ELISA检测细胞上清液中IL-1α、IL-6和TGF-β含量。结果显示,与对照组相比,MMP-7过表达组和M2型诱导组中Gal-3、MMP-7 mRNA表达显著上升(均P<0.01),(F4/80^(+))/CD206^(+)巨噬细胞百分比比值显著上升(均P<0.01),TGF-β浓度显著上升(均P<0.01),IL-1α、IL-6浓度显著下降(均P<0.01);M1型诱导组(F4/80^(+))/CD86^(+)巨噬细胞百分比比值显著上升(P<0.01),TGF-β浓度显著下降(P<0.01),IL-1α、IL-6浓度均显著增加(均P<0.01)。与M1型诱导组相比,M1型+Gal-3抑制剂组Gal-3 mRNA相对表达量显著下降(P<0.01),TGF-β浓度显著增加(P<0.01),而IL-1α、IL-6浓度显著下降(均P<0.01)。与M2型诱导组相比,M2型+Gal-3抑制剂组Gal-3 mRNA相对表达量显著降低(P<0.05),TGF-β浓度显著增加(P<0.01),而IL-1α、IL-6浓度显著下降(均P<0.05)。上述研究证明,MMP-7可上调Gal-3表达,进而促进巨噬细胞向M2型极化,发挥抗炎作用。To explore the effect of matrix metalloproteinase 7(MMP-7)over-expression on macrophage polarization,galectin 3(Gal-3)and inflammatory factors secretions,THP-1 cells were induced to macrophages and transfected with MMP-7 over-expressing plasmid or empty vector.The cells were divided into control group,MMP-7 over-expressing group,MMP-7 NC group,M1-polarized group,and M2-polarized group.mRNA expression levels of Gal-3 and MMP-7 were quantified by qPCR.The percentages of F4/80~+,CD86~+,and CD206~+macrophages were detected by flow cytometry.The expressions of inflammatory cytokines IL-1α,IL-6,and TGF-βwere detected by ELISA.After Gal-3 inhibitor treatment on M1-induced and M2-induced macrophages,the mRNA expression of Gal-3 in above macrophages was detected by qPCR,and the levels of IL-1α,IL-6,and TGF-βin the supernatant of cell cultures were detected by ELISA.The results showed that compared to the control group,the MMP-7 over-expressing group and the M2-polarized group showed significant increase in the expressions of Gal-3 and MMP-7 mRNA,in the percentage of(F4/80~+)/CD206~+macrophages(all with P<0.01),and in the concentrations of TGF-β(both P<0.01),while IL-1αand IL-6 levels showed significant decrease(both P<0.01).In the M1-polarized group,the percentage of(F4/80~+)/CD86~+macrophages increased significantly(P<0.01).The concentration of TGF-βdecreased,while the concentrations of IL-1αand IL-6 increased significantly(all with P<0.01).Compared to that of the M1-polarized group,the relative expression of Gal-3 mRNA in the M1+Gal-3 inhibitor group was significantly decreased(P<0.01),the concentration of TGF-βsignificantly increased(P<0.01),while the IL-1αand IL-6 levels significantly decreased(both P<0.01).Compared to those of the M2-polarized group,the relative expression of Gal-3 mRNA in M2+Gal-3 inhibitor group was significantly decreased(P<0.05),along with decreased IL-1αand IL-6 and increased TGF-βconcentrations(all with P<0.05).This study demonstrates that MMP-7 can activate Gal-3 expression to prom

关 键 词：基质金属蛋白酶7 巨噬细胞极化 半乳糖凝集素3 免疫应答 

分 类 号：R543.5[医药卫生—心血管疾病]