Berberine inhibits autophagy and promotes apoptosis of fibroblast-like synovial cells from rheumatoid arthritis patients through the ROS/mTOR signaling pathway.
10.12122/j.issn.1673-4254.2023.04.07
- Author:
Shiye ZONG
1
;
Jing ZHOU
2
;
Weiwei CAI
1
;
Yun YU
1
;
Ying WANG
1
;
Yining SONG
1
;
Jingwen CHENG
1
;
Yuhui LI
1
;
Yi GAO
1
;
Baihai WU
1
;
He XIAN
1
;
Fang WEI
1
Author Information
1. School of Pharmacy, Bengbu Medical College, Bengbu 233030, China.
2. Department of Pharmacy, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou 310007, China.
- Publication Type:Journal Article
- Keywords:
apoptosis;
autophagy;
berberine;
fibroblast-like synoviocytes;
mTOR;
reactive oxygen species;
rheumatoid arthritis
- MeSH:
Humans;
Synoviocytes;
Berberine/metabolism*;
Reactive Oxygen Species/metabolism*;
Tumor Necrosis Factor-alpha/metabolism*;
Hydrogen Peroxide/metabolism*;
Sincalide/metabolism*;
Cell Proliferation;
Arthritis, Rheumatoid/metabolism*;
Signal Transduction;
TOR Serine-Threonine Kinases/metabolism*;
Apoptosis;
Fibroblasts;
Autophagy;
Cells, Cultured
- From:
Journal of Southern Medical University
2023;43(4):552-559
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To evaluate the regulatory effect of berberine on autophagy and apoptosis balance of fibroblast-like synoviocytes (FLSs) from patients with in rheumatoid arthritis (RA) and explore the mechanism.
METHODS:The inhibitory effect of 10, 20, 30, 40, 50, 60, 70, and 80 μmol/L berberine on RA-FLS proliferation was assessed using CCK-8 method. Annexin V/PI and JC-1 immunofluorescence staining was used to analyze the effect of berberine (30 μmol/L) on apoptosis of 25 ng/mL TNF-α- induced RA-FLSs, and Western blotting was performed to detect the changes in the expression levels of autophagy- and apoptosis-related proteins. The cells were further treated with the autophagy inducer RAPA and the autophagy inhibitor chloroquine to observe the changes in autophagic flow by laser confocal detection of mCherry-EGFP-LC3B. RA-FLSs were treated with the reactive oxygen species (ROS) mimic H2O2 or the ROS inhibitor NAC, and the effects of berberine on ROS, mTOR and p-mTOR levels were observed.
RESULTS:The results of CCK-8 assay showed that berberine significantly inhibited the proliferation of RA-FLSs in a time- and concentration-dependent manner. Flow cytometry and JC-1 staining showed that berberine (30 μmol/L) significantly increased apoptosis rate (P < 0.01) and reduced the mitochondrial membrane potential of RA-FLSs (P < 0.05). Berberine treatment obviously decreased the ratios of Bcl-2/Bax (P < 0.05) and LC3B-II/I (P < 0.01) and increased the expression of p62 protein in the cells (P < 0.05). Detection of mCherry-EGFP-LC3B autophagy flow revealed obvious autophagy flow block in berberine-treated RA-FLSs. Berberine significantly reduced the level of ROS in TNF-α-induced RA-FLSs and upregulated the expression level of autophagy-related protein p-mTOR (P < 0.01); this effect was regulated by ROS level, and the combined use of RAPA significantly reduced the pro-apoptotic effect of berberine in RA-FLSs (P < 0.01).
CONCLUSION:Berberine can inhibit autophagy and promote apoptosis of RA-FLSs by regulating the ROS-mTOR pathway.
