基于 PI3K/Akt/mTOR信号通路探讨土家药铃铃菜干预类风湿关节炎的机制OA
Investigating the Mechanism of the Tujia Herbal Medicine Linglingcai in Rheumatoid Arthritis Intervention Based on the PI3K/Akt/mTOR Signaling Pathway
目的 从PI3K/Akt/mTOR信号通路的角度,探讨土家药铃铃菜(LLC)干预类风湿关节炎(RA)的药效作用机制.方法 将 36 只 SD 雄性大鼠随机分为正常组、模型组、雷公藤多苷组(10 mg·kg-1)及铃铃菜低(15 mg·kg-1)、中(30 mg·kg-1)、高(60 mg·kg-1)剂量组,共 6 组.除正常组以外,其它各组均采用牛Ⅱ型胶原皮下注射的方式建立大鼠类风湿关节炎(CIA)模型.造模成功后,各组按规定的剂量灌胃给药,连续给药 5 周.采用HE染色观察大鼠踝关节组织的病理变化;ELISA试剂盒检测血清中白细胞介素 1β(IL-1β)、白细胞介素6(IL-6)、干扰素α(TNF-α)的水平;RT-PCR法检测大鼠血清中IL-1β、IL-6、TNF-α的mRNA相对表达量;中药非靶标代谢组学(TIC)法检测铃铃菜的主要化学成分;网络药理学预测铃铃菜干预类风湿关节炎的可能机制;动物实验验证网络药理学预测信号通路的作用机制.结果 雷公藤多苷组及铃铃菜各剂量组大鼠关节肿胀评分和步态评分均降低(P＜0.01,P＜0.05);组织病理学检测显示细胞炎症浸润情况改善;血清IL-1β、IL-6、TNF-α的含量及mRNA表达下降(P＜0.01,P＜0.05).中药非靶标代谢组学显示铃铃菜共有 2 372 个化学成分.网络药理学结果显示,PI3K/Akt/mTOR信号通路可能是铃铃菜干预类风湿关节炎的通路,PI3K、Akt、mTOR可能是其干预类风湿关节炎的关键靶点.免疫组化(IHC)结果显示,铃铃菜可抑制大鼠踝关节细胞PI3K/Akt/mTOR通路的磷酸化活性.结论 铃铃菜可能通过抑制PI3K/Akt/mTOR信号通路磷酸化,减少关节炎症细胞的浸润,从而发挥抗类风湿关节炎的作用.
Objective To investigate the pharmacodynamic mechanism of the Tujia herbal medicine Linglingcai(LLC)in the intervention of rheumatoid arthritis(RA)from the perspective of the PI3K/Akt/mTOR signaling pathway.Methods A total of 36 male SD rats were randomly divided into six groups:normal control,model control,tripterygium glycosides(10 mg·kg-1),and LLC low-(15 mg·kg-1),medium-(30 mg·kg-1),and high-dose(60 mg·kg-1)groups.Except for the normal control group,a rat model of collagen-induced arthritis(CIA)was established by subcutaneous injection of bovine type Ⅱ collagen in all other groups.After successful modeling,the rats were administered the respective treatments by gavage for five consecutive weeks.Pathological changes in the rat ankle joint tissues were assessed using HE staining.Serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-alpha(TNF-α)were measured using ELISA kits.The relative mRNA expression levels of IL-1β,IL-6,and TNF-α in rat serum were determined by RT-PCR.The major chemical components of LLC were identified using untargeted metabolomics(TIC).Network pharmacology was employed to predict the potential mechanisms by which LLC intervenes in RA.Subsequently,animal experiments were conducted to validate the mechanism of the signaling pathway predicted by network pharmacology.Results Compared with the model group,the joint swelling scores and gait scores of rats in the tripterygium glycosides group and all LLC dose groups were significantly reduced(P＜0.01,P＜0.05).Histopathological examination showed amelioration of inflammatory cell infiltration.Serum levels and mRNA expression of IL-1β,IL-6,and TNF-α were significantly decreased(P＜0.01,P＜0.05).Untargeted metabolomics identified a total of 2 372 chemical components in LLC.Network pharmacology results indicated that the PI3K/Akt/mTOR signaling pathway might be the primary pathway through which LLC intervenes in RA,with PI3K,Akt,and mTOR identified as potential key targets.Immunohistochemistry(IHC)results demonstrated that LLC inhibited the phosphorylation activity of the PI3K/Akt/mTOR pathway in rat ankle joint cells.Conclusion LLC may exert its anti-rheumatoid arthritis effects by inhibiting the phosphorylation of the PI3K/Akt/mTOR signaling pathway,thereby reducing the infiltration of inflammatory cells into the joints.
王怡然;韩文轩;杨文婧;刘浩然;吴昊;董倩男
湖北恩施学院,湖北 恩施 445000湖北恩施学院,湖北 恩施 445000湖北恩施学院,湖北 恩施 445000湖北恩施学院,湖北 恩施 445000湖北恩施学院,湖北 恩施 445000湖北恩施学院,湖北 恩施 445000
医药卫生
铃铃菜土家药类风湿关节炎网络药理学PI3K/Akt/mTOR信号通路大鼠
LinglingcaiTujia herbal medicinerheumatoid arthritisnetwork pharmacologyPI3K/Akt/mTOR signaling pathwayrats
《中药新药与临床药理》 2026 (6)
1017-1027,11
国家自然科学基金资助项目(81960776)恩施州科技计划青年人才类项目(D20220034)湖北恩施学校级科研项目(KYJZ202307).
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