Qianggu Kangshu Formula attenuates osteoclast differentiation in rheumatoid arthritis by inhibiting the HIF-1α/BNIP3 autophagy signaling pathway.
10.12122/j.issn.1673-4254.2025.07.05
- Author:
Weiyi LI
1
;
Lu JIANG
2
;
Zongxing ZHANG
1
;
Dan CHEN
1
;
Zhuoma BAO
1
;
Li HUANG
3
;
Lin YUAN
1
Author Information
1. Hubei Provincial Key Laboratory of Rheumatic Disease Occurrence and Intervention, Hubei Minzu University, Enshi 445000, China.
2. Department of General Medicine, Shizhu County Hospital of Traditional Chinese Medicine, Chongqing 409100, China.
3. Department of Rheumatology, Enshi Huiyi Hospital of Rheumatic Diseases, Enshi 445000, China.
- Publication Type:Journal Article
- Keywords:
HIF-1α/BNIP3 signaling pathway;
Qianggu Kangshu Formula;
autophagy;
osteoclasts;
rheumatoid arthritis bone destruction
- MeSH:
Animals;
Drugs, Chinese Herbal/pharmacology*;
Osteoclasts/drug effects*;
Autophagy/drug effects*;
Mice;
Signal Transduction/drug effects*;
Rats;
Cell Differentiation/drug effects*;
Arthritis, Rheumatoid/pathology*;
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism*;
RAW 264.7 Cells;
Membrane Proteins/metabolism*;
Mitochondrial Proteins
- From:
Journal of Southern Medical University
2025;45(7):1389-1396
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:To investigate the effect of Qianggu Kangshu Formula (QGKSF) for alleviating osteoclast differentiation in rheumatoid arthritis and the underlying mechanism.
METHODS:RAW264.7 cells cultured under hypoxic conditions were treated with RANKL to induce osteoclast differentiation and incubated with normal rat serum or sera from rats medicated with methotrexate (MTX) or QGKSF at low and high doses. Cell viability, TRAP-positive multinucleated cells and F-actin ring formation in the treated cells were assessed with CCK-8 assay, TRAP staining and Phalloidin staining, respectively. Autophagy and autophagosomes in the cells were observed with MDC staining and transmission electron microscopy. ELISA was used to measure IL-6 and TNF-α levels in the culture supernatant, and the expressions of HIF-1α, BNIP3, Bcl-2, Beclin1, LC3-I, LC3-II, P62 and TRAP mRNAs and proteins were analyzed using RT-qPCR and Western blotting.
RESULTS:In hypoxia- and RANKL-induced RAW264.7 cells treated with normal rat serum, significant increments of TRAP-positive cells and F-actin ring formation were observed with an enhanced autophagic fluorescence intensity and increased autophagosomes. Treatment of the induced cells with rat sera medicated with MTX and low- and high-dose QGKSF obviously reduced the TRAP-positive cells, F-actin rings and autophagosomes as well as the autophagic fluorescence intensity. RANKL treatment significantly increased IL-6 and TNF-α levels in RAW264.7 cells, which were obviously decreased by treatment with MTX- and QGKSF-medicated sera. RANKL also significantly increased the mRNA and protein expression levels of HIF-1α, BNIP3, Bcl-2, Beclin1, LC3 and TRAP and lowered P62 expressions, and these changes were effectively reversed by treatment with MTX- and QGKSF-medicated sera.
CONCLUSIONS:QGKSF attenuates RANKL-induced osteoclast differentiation in hypoxic RAW264.7 cells by inhibiting the HIF-1α/BNIP3 autophagy signaling pathway, suggesting its potential for treatment of bone destruction in rheumatoid arthritis.
