中药药理与临床2025，Vol.41Issue(2)：39-47,9.
异鼠李素通过调控P38 MAPK和AKT/MTOR通路改善大鼠类风湿关节炎
Isorhamnetin Improves Rheumatoid Arthritis in Rats by Regulating P38 MAPK and AKT/MTOR Pathways
摘要
Abstract
Objective:To investigate the effect of isorhamnetin on the progression of rheumatoid arthritis(RA)and its underlying mechanisms.Methods:A collagen-induced arthritis(CIA)rat model was established using type II collagen(CII),and RA fibroblast-like synoviocytes(RA-FLS)were induced by tumor necrosis factor-α(TNF-α)as a cell model of RA.Tripterygium glycosides were used as a positive control drug,and isorhamnetin was added to study its effects and mechanisms.The arthritis index was assessed based on the degree of joint redness and swelling.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology of synovial tissue in rats.Cell viability was measured using the Cell Counting Kit-8(CCK-8).Cell migration was assessed by the Transwell method in vitro.The levels of inflammatory factors interleukin-1β(IL-1β),IL-6,IL-10,monocyte chemoattractant protein-1(MCP-1),and macrophage inflammatory protein-1α(MIP-1α)in the cell supernatant were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of matrix metallo-proteinase-2(Mmp2),Mmp3,Mmp8,Mmp9,P38,protein kinase B(Akt),mammalian target of rapamycin(Mtor),and P65 were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The phosphorylation levels of P38,AKT,MTOR,and P65 were detected by Western blot.Results:Animal experiments showed that compared with the normal control group,the model control group showed significant swelling in the rats'foot joints,with a significant increase in the arthritis index(P<0.01).In the synovial tissues,massive infiltration of in-flammatory cells and synovial hyperplasia were observed,along with significantly increased phosphorylation levels of P38,AKT,MTOR,and P65(P<0.01).Compared with the results in the model control group,the degree of swelling in the foot joints of rats in the 9 mg/kg triptery-gium glycosides group and the 20 mg/kg isorhamnetin group was significantly reduced,with a significant decrease in the arthritis score(P<0.01).The infiltration of inflammatory cells and symptoms of synovial hyperplasia were significantly improved,and the phosphorylation levels of P38,AKT,MTOR,and P65 were significantly downregulated(P<0.01).Cell experiments showed that compared with the results in the normal control group,the proliferation activity and migration ability of RA-FLS in the model control group were significantly enhanced(P<0.05 or P<0.01),the levels of IL-1β,IL-6,MCP-1,and MIP-1α were significantly increased(P<0.01),and the IL-10 level was significant-ly decreased(P<0.01).The mRNA expression levels of Mmp2,Mmp3,Mmp8,and Mmp9 were significantly upregulated(P<0.01),along with the mRNA expression levels of P38,Akt,Mtor,and P65,and the phosphorylation levels of P38,AKT,MTOR,and P65(P<0.01).Com-pared with the results in the model control group,the proliferation activity and migration ability of RA-FLS were significantly reduced in the 0.030 mg/mL tripterygium glycosides group and the 0.094,0.188,and 0.375 mmol/L isorhamnetin groups(P<0.05 or P<0.01).The lev-els of IL-1β,IL-6,MCP-1,and MIP-1α in RA-FLS were significantly reduced in the 0.030 mg/mL tripterygium glycosides group and the 0.375 mmol/L isorhamnetin group(P<0.01),while the level of IL-10 was significantly increased(P<0.01).The mRNA expression levels of Mmp2,Mmp3,Mmp8,and Mmp9 were significantly downregulated(P<0.01),along with the mRNA expression levels of P38,Akt,Mtor,and P65,and the phosphorylation levels of P38,AKT,MTOR,and P65(P<0.05 or P<0.01).Conclusion:Isorhamnetin can inhibit the prolifera-tion,migration,and changes in inflammatory factors of RA-FLS,thereby alleviating RA symptoms in CIA rats.The mechanism involves the regulation of the P38 MAPK and AKT/MTOR signaling pathways.关键词
异鼠李素/类风湿关节炎/P38 丝裂原活化蛋白激酶/蛋白激酶B-雷帕霉素靶蛋白/增殖和迁移Key words
Isorhamnetin/Rheumatoid arthritis/P38 mitogen-activated protein kinase/Protein kinase B/mammalian target of rapamycin/Pro-liferation and migration引用本文复制引用
刘笑蓉,李硕夫,刘湘丹,龙雨青,周日宝..异鼠李素通过调控P38 MAPK和AKT/MTOR通路改善大鼠类风湿关节炎[J].中药药理与临床,2025,41(2):39-47,9.基金项目
湖南省自然科学基金项目(编号:2022JJ80086、2023JJ60342) （编号:2022JJ80086、2023JJ60342）
湖南省卫健委科学研究项目(编号:D202302078705) （编号:D202302078705）
湖南省教育厅科学研究项目(编号:23B0385) （编号:23B0385）
湖南省大学生创新创业训练计划项目(编号:2022-5313) （编号:2022-5313）
湖南省中医药管理局科研计划项目编号:(A2024002、B2023150) （A2024002、B2023150）
湖南中医药大学2023本科生科研创新基金项目编号:(校行科字[2023]7) （校行科字[2023]7）
湖南中医药大学校级科研项目编号:(2023-27) （2023-27）
湖南中医药大学青苗计划(校行人字[2017]25) （校行人字[2017]25）
湖南中医药大学重点学科中药学科(校行发规字[2023]2号) （校行发规字[2023]2号）
2020年湖南省一流本科专业建设点(湘教通[2020]248号):中药资源与开发 （湘教通[2020]248号）
2021年国家级一流本科专业建设点(教高厅函[2021]7号). （教高厅函[2021]7号）
