基于非靶向代谢组学探究草乌不同炮制品抗类风湿性关节炎的机制

Exploration on the mechanism of different processed products of Aconitum kusnezoffii Reichb in the treatment of rheumatoid arthritis based on non-targeted metabolomics

目的:利用超高效液相色谱串联傅里叶变换质谱(UHPLC-FT-MS)技术探讨草乌及其炮制品对类风湿性关节炎模型大鼠血清代谢物的影响,并探究其潜在抗炎机制。方法:建立Ⅱ型胶原诱导性关节炎(CIA)大鼠模型,造模成功后,灌胃给予生草乌、《中华人民共和国药典》(以下简称《药典》)制草乌和诃子制草乌5周。监测大鼠的一般情况、足肿胀度和关节炎指数,并使用酶联免疫吸附法(ELISA)检测TNF-α、IL-6和IL-1β的水平。非靶向代谢组学技术分析大鼠血清中代谢物轮廓变化,寻找可能的差异代谢物,并对差异代谢物做进一步通路富集分析。结果:与模型组相比,草乌及其炮制品治疗可减轻CIA大鼠的足肿胀,降低关节炎指数,减少炎症因子TNF-α、IL-6和IL-1β的分泌。造模后,血清中50种代谢物发生了显著改变(P<0.05),可视为潜在生物标志物来表征CIA大鼠的代谢紊乱。其中生草乌主要参与氨基酸代谢、脂质代谢和其它氨基酸的代谢;诃子制草乌主要参与脂质代谢和碳水化合物代谢;《药典》制草乌主要参与氨基酸代谢、脂质代谢、碳水化合物代谢、辅因子和维生素的代谢。结论:草乌及其炮制品可通过调节血清代谢物,改善CIA大鼠氨基酸代谢、脂质代谢及甘露糖代谢紊乱来延缓CIA大鼠炎症的发生发展,为草乌及其炮制品抗类风湿性关节炎的作用机制做出了新的解释。

Objective:To investigate the effects of Aconitum kusnezoffii Reichb and its processed products on serum metabolites in rats with rheumatoid arthritis using ultra-performance liquid chromatography tandem Fourier transform mass spectrometry(UHPLC-FT-MS),and to explore its potential anti-inflammatory mechanism.Methods:The collagen-induced arthritis(CIA)rat model was established.After successful establishment of the model,the rats were orally treated with SCW,YDZCW and HZZCW for 5 weeks.The general condition,degree of foot swelling and arthritis index of the rats were monitored.Levels of TNF-α,IL-6 and IL-1βwere detected by using ELISA.Non-targeted metabolomics technology was used to analyze changes in the serum metabolite profile in rats,identify potential differential metabolites,and perform pathway enrichment analysis for these metabolites.Results:Compared to the model group,treatment with Aconitum kusnezoffii Reichb and its processed products reduced foot swelling,arthritis index,as well as secretion of inflammatory factors TNF-α,IL-6 and IL-1βin CIA rats.After modeling,CIA resulted in significant changes(P<0.05)in 50 serum metabolites which can be considered as potential biomarkers characterizing metabolic disorders in CIA rats.SCW was mainly involved in amino acid,lipid and other related metabolic processes.HZZCW was mainly involved in lipid and carbohydrate metabolism.YDZCW was mainly involved in amino acid,lipid,carbohydrate,cofactor and vitamin metabolism.Conclusion:Aconitum kusnezoffii Reichb and its processed products exhibit anti-inflammatory effects in CIA rats by modulating serum metabolites and ameliorating dysregulation of amino acid,lipid,and mannose metabolism,thereby providing a novel mechanistic insight into their therapeutic potential for rheumatoid arthritis.