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
- VernacularTitle:缺氧微环境下补阳还五汤通过抑制BNIP3-PI3K/Akt通路抑制类风湿关节炎滑膜成纤维细胞的线粒体自噬
- Author:
Junping ZHAN
1
;
Shuo HUANG
;
Qingliang MENG
;
Wei FAN
;
Huimin GU
;
Jiakang CUI
;
Huilian WANG
Author Information
1. 河南省中医院//河南中医药大学第二附属医院风湿科,河南郑州 450002
- Publication Type:Journal Article
- Keywords:
hypoxia;
Buyang Huanwu Decoction;
rheumatoid arthritis;
synovial fibroblast;
BNIP3-PI3K/Akt;
mitochondrial autophagy
- From:
Journal of Southern Medical University
2025;45(1):35-42
- CountryChina
- Language:Chinese
-
Abstract:
Objective 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.Conclusion BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.
