Arecoline induces activation of human oral fibroblasts by promoting macrophage secretion of exosomes containing miR-155-5p.
10.12122/j.issn.1673-4254.2023.01.08
- Author:
Yong Qi HUANG
1
;
Wei YU
1
;
Yue Hua YOU
2
Author Information
1. School of Stomatology, Southern Medical University, Guangzhou 510515, China.
2. Department of Stomatology, Longhua People's Hospital Affiliated to Southern Medical University, Shenzhen 518109, China.
- Publication Type:Journal Article
- Keywords:
arecoline;
exosome;
macrophages;
oral fibroblasts;
oral submucosal fibrosis
- MeSH:
Humans;
Exosomes;
Arecoline/pharmacology*;
Collagen Type I;
Fibroblasts;
Macrophages;
MicroRNAs
- From:
Journal of Southern Medical University
2023;43(1):60-67
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the mechanism by which arecoline regulates the level of miR-155-5p in macrophage-secreted exosomes to induce the transformation of human oral mucosal fibroblasts (HOMFs) into fibroblast phenotype.
METHODS:Exosomes were harvested from human monocytic cell line THP-1 with or without arecoline treatment. The effects of arecoline-treated THP-1 cell culture supernatant (CS), THP-1-derived exosomes (EXO), exosome-depleted THP-1 cell supernatant (NES), miR-155-5p overexpression, and miR-155-5p inhibitor on migration ability of arecoline-treated HOMF cells were examined using Transwell migration assay. The polarization of THP-1 cells was detected using flow cytometry. DCFH-DA was used to detect the level of oxidative stress in the cells with different treatments. The mRNA and protein expressions of α- SMA, type I collagen and SOCS1 in the cells were detected with qRT-PCR and Western blotting.
RESULTS:Flow cytometry showed that arecoline-treated THP-1 cells exhibited obvious polarization from M0 to M1. Both the supernatant and exosomes from arecoline-treated THP-1 cells significantly enhanced the migration ability of HOMF cells, increased intracellular oxidative stress, up-regulated the expressions of miR-155- 5p and the mRNA and protein levels of α-SMA and type I collagen, and lowered the mRNA and protein expressions of SOCS1. In HOMF cells treated with exosomes from arecoline- treated THP-1 cells, overexpression of miR-155-5p significantly enhanced cell migration ability and increased cellular expressions of α-SMA and type I collagen, and miR-155-5p inhibitor caused the opposite changes.
CONCLUSION:Arecoline can up-regulate miR-155-5p expression in THP-1 cells and inhibit the expression of SOCS1 protein in HOMF cells via the exosome pathway, thus promoting the fibrotic phenotype transformation of HOMF cells.
