中国中西医结合急救杂志2026，Vol.33Issue(2)：209-216,8.DOI:10.3969/j.issn.1008-9691.2026.02.014
黄芪甲苷通过SIRT1/AP-1信号轴调控软骨细胞氧化应激的机制研究
Study on the mechanism of astragaloside Ⅳ regulating oxidative stress in chondrocytes via the sirtuin 1/activator protein-1 signaling axis
摘要
Abstract
Objective To explore the protective effect of astragalosideⅣ(AS-Ⅳ)on oxidative damage model induced by interleukin-1β(IL-1β)in SW1353 cells and its underlying mechanisms.Methods The SW1353 cells were treated with 100 μg/L IL-1β for 48 hours to construct an in vitro cell model of oxidative damage.The cells were randomly divided into six groups:blank control,model,AS-Ⅳ,sirtuin 1(SIRT1)-knockdown,activator protein-1(AP-1)-overexpression,and SIRT1-knockdown+AP-1-overexpression.The optimal concentration and time for IL-1β and the optimal concentration and time for AS-Ⅳ in the treatment of osteoarthritis(OA)were screened using the cell counting kit-8(CCK-8)assay.SIRT1 knockdown and AP-1 overexpression were achieved by lentiviral transfection.The contents of malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)in each group of cells were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expressions of AP-1,SIRT1,Bax,Bcl-2,and caspase-3 were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blotting,respectively.Results The contents of CAT(kU/L:7.64±2.36 vs.26.98±2.05),SOD(kU/L:25.92±1.78 vs.39.63±2.51),and GSH-Px(kU/L:176.70±8.36 vs.261.40±12.71)in the model group were significantly lower than those in the blank control group(all P<0.05),while the content of MDA was significantly higher(μmol/L:13.52±2.88 vs.6.24±0.37,P<0.05).After the addition of AS-Ⅳ,compared with the model group,the contents of CAT(kU/L:24.82±2.49 vs.7.64±2.36),SOD(kU/L:38.56±2.25 vs.25.92±1.78),and GSH-Px(kU/L:231.40±20.11 vs.176.70±8.36)in the AS-Ⅳ group were significantly increased(both P<0.05),and the content of MDA(μmol/L:7.12±1.00 vs.13.52±2.88,P<0.05)was significantly decreased.Moreover,AS-Ⅳ could effectively inhibit the apoptosis of SW1353 cells,down-regulate the protein expression of AP-1(AP-1/Tubulin:0.39±0.03 vs.1.11±0.06,P<0.05),and up-regulate the protein expression of SIRT1(SIRT1/Tubulin:1.34±0.11 vs.1.06±0.12,P<0.05).After reducing SIRT1 knockdown,AP-1 overexpression,and combined SIRT1 knockdown with AP-1 overexpression,compared with the AS-Ⅳ group,the contents of CAT(kU/L:5.32±1.22,5.93±2.11,4.13±1.93 vs.24.82±2.49),SOD(kU/L:25.19±1.18,26.02±2.04,21.02±1.56 vs.38.56±2.25),and GSH-Px(kU/L:181.40±12.71,175.50±10.40,148.00±16.64 vs.231.40±20.11)in the SIRT1 group,AP-1 group,and SIRT1+AP-1 group were significantly decreased(all P<0.05),while the MDA content increased(μmol/L:13.83±2.51,16.85±2.74,45.00±1.66 vs.7.12±1.00,all P<0.05),and the mRNA and protein expression levels of AP-1,Bax,and caspase-3 were significantly increased[AP-1 mRNA expression(2-ΔΔCt):4.93±0.25,5.30±0.57,6.10±0.18 vs.1.53±0.42,AP-1 protein expression(AP-1/Tubulin):1.11±0.04,1.18±0.10,1.60±0.09 vs.0.39±0.03;Bax mRNA expression(2-ΔΔCt):4.60±0.43,4.76±0.63,6.16±0.53 vs.1.49±0.45,Bax protein expression(Bax/Tubulin):0.80±0.11,0.92±0.06,1.48±0.10 vs.0.43±0.08;caspase-3 mRNA expression(2-ΔΔCt):7.27±0.90,7.12±1.09,9.38±0.58 vs.1.20±0.35,caspase-3 protein expression(caspase-3/Tubulin):0.75±0.11,0.79±0.06,1.02±0.11 vs.0.48±0.04,all P<0.05],while the mRNA and protein expression levels of SIRT1 and Bcl-2 were significantly decreased(SIRT1 mRNA expression(2-ΔΔCt):0.64±0.09,0.63±0.05,0.28±0.06 vs.0.86±0.05,SIRT1 protein expression(SIRT1/Tubulin):0.94±0.05,0.97±0.07,0.26±0.06 vs.1.34±0.11;Bcl-2 mRNA expression(2-ΔΔCt):0.38±0.04,0.43±0.05,0.18±0.08 vs.0.70±0.09,Bcl-2 protein expression(Bcl-2/Tubulin):0.76±0.06,0.80±0.07,0.34±0.06 vs.1.15±0.08,all P<0.05).Conclusion AS-Ⅳ can protect SW1353 cells from oxidative damage induced by IL-1β and alleviate cell apoptosis,the mechanism of its action may be related to its activation of the SIRT1/AP-1 signaling pathway.关键词
黄芪甲苷/氧化损伤/凋亡/沉默信息调节因子 1/激活蛋白-1 信号通路Key words
Astragaloside Ⅳ/Oxidative damage/Apoptosis/Sirtuin 1/activator protein-1 signaling pathway引用本文复制引用
王桂宇,王建国,赖林,蒲萌,贾卓樾,刘琪..黄芪甲苷通过SIRT1/AP-1信号轴调控软骨细胞氧化应激的机制研究[J].中国中西医结合急救杂志,2026,33(2):209-216,8.基金项目
山西省中医药管理局山西中医药大学方药配伍及功用重点研究室开放课题(zyyyis2024023) （zyyyis2024023）
山西中医药大学科技创新能力培育计划(2024PY-JL-9-01,2024PY-JL-9-02) （2024PY-JL-9-01,2024PY-JL-9-02）
山西中医药大学杏林英才计划项目(2025XJ01) The Open Research Project of the Key Laboratory of Formula and Function of Traditional Chinese Medicine,Shanxi University of Chinese Medicine and Shanxi Provincial Administration of Traditional Chinese Medicine(zyyyis2024023) （2025XJ01）
Scientific and Technological Innovation Capacity Cultivation Program of Shanxi University of Chinese Medicine(2024PY-JL-9-01,2024PY-JL-9-02) （2024PY-JL-9-01,2024PY-JL-9-02）
Xinglin Talent Project of Shanxi University of Chinese Medicine(2025XJ01) （2025XJ01）
