miR-148b inhibits M2 polarization of LPS-stimulated macrophages by targeting DcR3.
10.3760/cma.j.cn112150-20220808-00794
- VernacularTitle:miR-148b 靶向调控DcR3抑制LPS诱导的巨噬细胞极化
- Author:
Li Yuan YANG
1
;
Xiao Li LOU
1
;
Yue WANG
1
;
Yan Qiang HOU
1
Author Information
1. Department of Clinical Laboratory, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (Preparatory stage), Shanghai 201600, China.
- Publication Type:Journal Article
- MeSH:
Humans;
Interleukin-10;
Lipopolysaccharides/pharmacology*;
Macrophages;
MicroRNAs/genetics*;
Receptors, Tumor Necrosis Factor, Member 6b/metabolism*;
Tumor Necrosis Factor-alpha
- From:
Chinese Journal of Preventive Medicine
2023;57(8):1231-1237
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
