Buyang Huanwu Decoction reduces mitochondrial autophagy in rheumatoid arthritis synovial fibroblasts in hypoxic culture by inhibiting the BNIP3-PI3K/Akt pathway.
10.12122/j.issn.1673-4254.2025.01.05
- Author:
Junping ZHAN
1
;
Shuo HUANG
2
;
Qingliang MENG
1
;
Wei FAN
1
;
Huimin GU
1
;
Jiakang CUI
1
;
Huilian WANG
1
Author Information
1. Department of Rheumatology, Henan Provincial Hospital of Traditional Chinese Medicine// Second Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450002, China.
2. College of Orthopedics and Traumatology, Henan University of Traditional Chinese Medicine, Zhengzhou 450008, China.
- Publication Type:Journal Article
- Keywords:
BNIP3-PI3K/Akt;
Buyang Huanwu Decoction;
hypoxia;
mitochondrial autophagy;
rheumatoid arthritis;
synovial fibroblast
- MeSH:
Drugs, Chinese Herbal/pharmacology*;
Arthritis, Rheumatoid/pathology*;
Animals;
Signal Transduction/drug effects*;
Proto-Oncogene Proteins c-akt/metabolism*;
Autophagy/drug effects*;
Humans;
Fibroblasts/cytology*;
Rats, Wistar;
Membrane Proteins/metabolism*;
Rats;
Phosphatidylinositol 3-Kinases/metabolism*;
Mitochondria/metabolism*;
Cells, Cultured;
Proto-Oncogene Proteins/metabolism*;
Apoptosis/drug effects*;
Cell Hypoxia;
Synovial Membrane/cytology*;
Male;
Mitochondrial Proteins
- From:
Journal of Southern Medical University
2025;45(1):35-42
- CountryChina
- Language:English
-
Abstract:
OBJECTIVES:To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction (BYHWT) on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients (FLS-RA) cultured under a hypoxic condition.
METHODS:Forty normal Wistar rats were randomized into two groups (n=20) for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera. FLS-RA were cultured in routine condition or exposed to hypoxia (10% O2) for 24 h wigh subsequent treatment with IL-1β, followed by treatment with diluted BYHWT-medicated serum (5%, 10% and 20%) or control serum. AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells, and the changes in ROS, ATP level, mitochondrial membrane potential and Ca2+ homeostasis were analyzed. The changes in mRNA and protein expressions of BNIP3, PI3K and AKT and mRNA expressions of LC3, Beclin-1 and P62 were detected using RT-qPCR and Western blotting.
RESULTS:Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure. The treatment significantly decreased T-AOC concentration, increased ROS production, autophagosome formation and ATPase levels, and lowered mitochondrial membrane potential and Ca2+ level in the cells. In IL-1β-induced FLS-RA with hypoxic exposure, treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression, decreased the protein expressions of PI3K and AKT, increased the mRNA expressions of BNIP3 and P62, and lowered the mRNA expressions of PI3K, AKT, LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression. The cells treated with 5% and 10% BYHWT-medicated serum showed no significant changes in LC3 expression.
CONCLUSIONS:BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.
