miR-190a-5p靶向C/EBPα-PU.1通路促进骨髓巨噬细胞M1型向M2型极化  

miR-190a-5p targeting the C/EBPα-PU.1 pathway promotes the polarization of bonemarrow-derived macrophages from M1-type to M2-type

作　　者：杨磊 薛松[1] 王亚祥 连锋[1] YANG Lei;XUE Song;WANG Yaxiang;LIAN Feng(Department of Cardiovascular Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200127,P.R.China)

机构地区：[1]上海交通大学医学院附属仁济医院心血管外科,上海200127

出　　处：《中国胸心血管外科临床杂志》2022年第8期1042-1048,共7页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金(81873461)。

摘　　要：目的探讨miR-190a-5p对脂多糖诱导的骨髓巨噬细胞(bone-marrow-derived macrophage,BMDM)向M1、M2型极化的影响。方法以细菌脂多糖诱导的BMDM(M1型)为M1组,阴性对照miRNA模拟物干预巨噬细胞M1型为NC组,miR-190a-5p模拟物干预巨噬细胞M1型为miR-190a-5p组。显微镜下观察巨噬细胞形态学改变,流式细胞仪检测M2型巨噬细胞(CD206+,F4/80)占整体比例。利用荧光定量PCR检测与巨噬细胞极化相关的精氨酸酶-1(arginase 1,Arg1)、一氧化氮合酶(inducible nitric oxide synthase,iNOS)、肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)、靶基因CCAAT/增强子结合蛋白-α(CCAAT enhancer-binding proteinα,C/EBPα)、PU.1的mRNA表达水平,验证C/EBPα、PU.1是否为miR-190a-5p的潜在靶基因。Western blotting实验检测通路蛋白C/EBPα、PU.1的表达情况。结果miR-190a-5p模拟物干预巨噬细胞M1型后,巨噬细胞触角伸长,呈长索状M2型细胞形态特征。miR-190a-5p模拟物干预巨噬细胞M1型24 h,M2型巨噬细胞增加。miR-190a-5p模拟物干预巨噬细胞M1型对mRNA表达水平的影响:iNOS和TNF-α的表达显著降低(P<0.05),M2巨噬细胞标志Arg1的表达显著升高(P<0.05),靶基因C/EBPα、PU.1的mRNA表达水平显著降低(P<0.05)。Western blotting实验结果显示,过表达miR-190a-5p明显抑制C/EBPα、PU.1蛋白表达,miR-190a-5p抑制剂促进C/EBPα、PU.1蛋白表达(P<0.05)。结论miR-190a-5p可促进BMDM从M1型极化为M2型。Objective To investigate the effect of miR-190a-5p on the polarization of bone-marrow-derived macrophage(BMDM)induced by lipopolysaccharides to M1-and M2-types.Methods BMDM(M1-type)induced by bacterial lipopolysaccharide was a M1 group.The macrophage M1-type interfered with negative control miRNA mimics was a NC group.miR-190a-5p mimics interfered with the M1-type of macrophages in the miR-190a-5p group.Morphological changes of macrophages were observed under a microscope,and the proportion of M2-type macrophages(CD206+,F4/80)was detected by flow cytometry.The mRNA expression levels of argininase-1(Arg1),inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),target gene C/EBPαand PU.1 were detected by fluorescence quantitative PCR to verify whether C/EBPαand PU.1 were potential target genes of miR-190a-5p.The expression of pathway proteins C/EBPαand PU.1 were detected by Western blotting.Results After miR-190a-5p mimics interfered with macrophage M1-type,the antenna of macrophages elongated and showed long cord M2-type cell morphological characteristics.miR-190a-5p mimics interfered with M1-type macrophages for 24 h,and the percentage of M2-type macrophages increased significantly(P<0.05).Effects of miR-190a-5p simulator on mRNA expression levels of M1-type macrophages included:the expression of iNOS and TNF-αwas significantly decreased(P<0.05),the expression of Arg1 marked by M2 macrophages was significantly increased(P<0.05),and the mRNA expression levels of target genes C/EBPαand PU.1 were significantly decreased(P<0.05).Western blotting results showed that the overexpression of miR-190a-5p significantly inhibited the protein expressions of C/EBPαand PU.1,while the miR-190a-5p inhibitor increased the expressions of both proteins.Conclusion miR-190a-5p can promote the polarization of BMDM from M1-type to M2-type.

关 键 词：miR-190a-5p 巨噬细胞 C/EBPΑ 极化 实验 

分 类 号：R392[医药卫生—免疫学]