miR-190a-5p targeting the C/EBPα-PU.1 pathway promotes the polarization of bone-marrow-derived macrophages from M1-type to M2-type
- VernacularTitle: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
- Author:
Lei YANG
1
;
Song XUE
1
;
Yaxiang WANG
1
;
Feng LIAN
1
Author Information
1. Department of Cardiovascular Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, P. R. China
- Publication Type:Journal Article
- Keywords:
miR-190a-5p;
macrophages;
C/EBPα;
polarization;
experiment
- From:
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery
2022;29(08):1042-1048
- CountryChina
- Language:Chinese
-
Abstract:
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/EBPα and PU.1 were detected by fluorescence quantitative PCR to verify whether C/EBPα and PU.1 were potential target genes of miR-190a-5p. The expression of pathway proteins C/EBPα and 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-α was 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/EBPα and 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/EBPα and 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.
