西南医科大学学报2025，Vol.48Issue(5)：478-485,8.DOI:10.3969/j.issn.2096-3351.2025.05.007
血根碱通过激活LKB1/AMPK/SIRT1通路抑制内质网应激和细胞衰老以改善骨关节炎
Sanguinarine Inhibits Endoplasmic Reticulum Stress and Cell Senescence through Activation of the LKB1/AMPK/SIRT1 Pathway to Ameliorate Osteoarthritis
摘要
Abstract
Objective The aim of this study was to investigate the role of Sanguinarine(SG)in the progression of osteoarthritis(OA)and to elucidate the possible mechanisms involved.Methods Primary mouse knee articular chondrocytes were isolated and treated with SG,tert-butyl hydroperoxide(TBHP),TBHP+SG,TBHP+SG+thapsigargin(TG),or TBHP+SG+liver kinase B1(LKB1)siRNA.Cell viability,apoptosis,ROS level and β-gal activity were detected by CCK-8,flow cytometry,DCFH-DA probe and β-galactosidase(β-gal)staining,respectively.The mRNA levels of senescence-associated secretory phenotype(SASP)was detected by RT-qPCR and LKB1/AMP-activated protein kinase(AMPK)/sirtuin 1(SIRT1)pathway related protein expressions were measured by Western blotting.Mice were divided into:the Sham group,the Sham+SG(1.25 μM,10 μL)group,the OA group,and the OA+SG(0.8/1.25 μM,10 μL)group,and cartilage loss and apoptosis were assessed using safranin O-fast green and TUNEL stain-ing.Results Concentration screening showed that the optimal concentration for SG treatment of chondrocytes was 1.25 μM.SG treat-ment reversed the TBHP-induced decrease in cell viability,and increase in apoptosis,ECM degradation,ROS generation,expression of senescence-associated proteins(such as cyclin dependent kinase inhibitor 1A(p21)and cyclin dependent kinase inhibitor 2A(p16)),mRNA expression of SASPs(such as monocyte chemoattractant protein-1(MCP-1),matrix metalloproteinase 3(MMP3),and C-X-C motif chemokine ligand 10(Cxcl10),and β-gal activity.TG treatment could reverse the above changes.SG treatment effec-tively neutralized the TBHP-induced up-regulation of ER stress-related proteins(such as glucose regulated protein 78kD(GRP78)and C/EBP Homologous Protein(CHOP))and reduced phosphorylation levels of LKB1 and AMPK and protein level of SIRT1.Silenc-ing of LKB1 resisted the inhibitory effect of SG on ER stress,suggesting that SG inhibited TBHP-induced ER stress by modulating the LKB1/AMPK/SIRT1 pathway.In vivo,SG administration attenuated chondrocyte apoptosis and cartilage loss in OA mice in a dose-dependent manner.Conclusion Sanguinarine inhibited endoplasmic reticulum stress by activating the LKB1/AMPK/SIRT1 signaling pathway,thereby alleviating ECM degradation,apoptosis,and senescence of chondrocytes induced by oxidative stress,and hindering the progression of osteoarthritis(OA)in mice,and providing a new theoretical basis for the treatment of OA.关键词
血根碱/LKB1/AMPK/SIRT1通路/ER应激/衰老/软骨细胞Key words
Sanguinarine/The LKB1/AMPK/SIRT1 pathway/Endoplasmic reticulum stress/Cell senescence/Chondrocytes分类
医药卫生引用本文复制引用
马运锋,韩小飞,曹玉净,史栋梁,陈志煌,储永良,王延涛..血根碱通过激活LKB1/AMPK/SIRT1通路抑制内质网应激和细胞衰老以改善骨关节炎[J].西南医科大学学报,2025,48(5):478-485,8.基金项目
河南省中医药科学研究专项(20-21ZY3013) （20-21ZY3013）
