时珍国医国药2025，Vol.36Issue(5)：827-837,11.DOI:10.70976/j.1008-0805.SZGYGY-2025-0505
基于AMPK/mTOR信号通路探讨龟鹿二仙胶及拆方对退变软骨细胞自噬机制影响的研究
Study on the effects of Guilu Erxin Jiao and its disassembled formulations on the autophagy mechanism of degengrative chondrocytes via the AMPK/mTOR signaling pathway
摘要
Abstract
Objective To investigate the effects of Guilu Erxian Jiao(GLJ)and its disassembled formulations on the autophagy mecha-nism of degenerative chondrocytes via the AMPK/mTOR signaling pathway.Methods Wild-type and mTOR KO mouse chondrocytes were cultured in vitro using enzyme digestion.IL-1β was used to induce F2 generation chondrocytes to establish a model of degenerative chondrocytes.The mouse chondrocytes were randomly divided into blank group,model group,Turtle and Deer group,Turtle Shell group,Deer Antler group,CKO group,Turtle and Deer+CKO group,Turtle Shell+CKO group,and Deer Antler+CKO group.Af-ter 24 h of serum intervention,chondrocyte viability was detected by CCK-8 method;immunofluorescence method was used to detect chondrocyte PARP expression;laser confocal microscopy was used to observe chondrocyte LC3B fluorescence intensity;qPCR method was used to detect chondrocyte AMPK,mTOR,COL2A1 mRNA expression;Western-Blot was used to detect chondrocyte AMPK,p-AMPK,mTOR,p-mTOR,COL2A1 protein expression.Results Compared with the model group,all intervention groups showed in-creased cell viability,enhanced LC3B fluorescence intensity,reduced PARP average fluorescence intensity,increased AMPK and COL2A1 protein and mRNA expression,and decreased mTOR and mRNA expressions(P<0.05).Compared with the GLJ group,the GLJ+CKO group exhibited higher cell viability,increased LC3B fluorescence intensity,decreased PARP fluorescence intensity,in-creased AMPK,COL2A1 and mRNA expressions,elevated p-AMPK protein expression,reduced mTOR and mRNA expressions,and decreased p-mTOR protein expression(P<0.05).Compared with the GB group,the LJ group exhibited higher cell viability,reduced LC3B fluorescence intensity,increased PARP fluorescence intensity,decreased AMPK and mRNA expression,increased COL2A1 and mRNA expression,decreased p-AMPK protein expression,and increased mTOR and mRNA expressions(P<0.05).The GB+CKO group showed enhanced cell viability,increased LC3B fluorescence intensity,decreased PARP fluorescence intensity,elevated AMPK and COL2A1 protein and mRNA expression,increased p-AMPK protein expression,and reduced mTOR protein,mTOR mRNA,and p-mTOR protein expressions(P<0.05).Compared with the LJ group,the LJ+CKO group showed improved cell viability,increased LC3B fluorescence intensity,decreased PARP fluorescence intensity,elevated AMPK and COL2A1 protein and mRNA expression,in-creased p-AMPK protein expression,and reduced mTOR protein,mTOR and mRNA,and p-mTOR protein expressions(P<0.05).Conclusion Guiban enhances chondrocyte autophagy and inhibits apoptosis,while Lujiao promotes chondrocyte proliferation and extra-cellular matrix(ECM)synthesis.GLJ demonstrates superior efficacy compared to Guiban and Lujiao,with its mechanism linked to the AMPK/mTOR pathway activation.关键词
龟鹿二仙胶/软骨细胞/AMPK/mTOR信号通路/自噬/凋亡Key words
Guilu Erxian Jiao/Chondrocytes/AMPK/mTOR signaling pathway/Autophagy/Apoptosis分类
医药卫生引用本文复制引用
蔡玮俊,吴伟欣,郑珍萍,钦洋溢,顾富城,杨美鑫,耿秋东,王和鸣,李楠..基于AMPK/mTOR信号通路探讨龟鹿二仙胶及拆方对退变软骨细胞自噬机制影响的研究[J].时珍国医国药,2025,36(5):827-837,11.基金项目
福建省自然科学基金(2022J01364) （2022J01364）
福建省财政专项项目(X2022002) （X2022002）
国家自然科学基金面上项目(81973880) （81973880）
