海南医科大学学报2025，Vol.31Issue(23)：1823-1832,10.DOI:10.13210/j.cnki.jhmu.20250327.004
基于网络药理学和非靶向代谢组学探究异丁酰紫草素抗银屑病的作用机制
Mechanism of isobutyrylshikonin against psoriasis based on network pharmacology and untargeted metabolomics
摘要
Abstract
Objective:To explore the potential mechanism of isobutyrylshikonin in the treatment of psoriasis by combining net-work pharmacology and untargeted metabolomics techniques.Methods:Network pharmacology was employed to screen the tar-gets and pathways of isobutyrylshikonin in psoriasis treatment,followed by molecular docking verification.RAW264.7 cells and HaCaT cells were used as the research objects in vitro.The anti-inflammatory activity of isobutyrylshikonin was investigated using a lipopolysaccharide(LPS)-induced inflammatory model of M1 macrophages(pro-inflammatory)and an interleukin-4(IL-4)-in-duced polarization model of M2 macrophages(anti-inflammatory).RT-qPCR was conducted to detect the expression of TNF-α,iNOS,CXCL9,CXCL10 in M1 macrophages,and CD206,TGF-β,Arg-1,IL-10 in M2 macrophages.Nitric oxide(NO)and reactive oxygen species(ROS)levels in macrophages were measured by Griess regent method and fluorescence staining method.Western blot was used to detect the expression levels of proteins related to the JAK/STAT signaling pathway.Immunofluores-cence was employed to investigate the nuclear translocation of p-STAT3 protein.UPLC-Q-TOF/MS technology was utilized to detect the differential metabolites and metabolic pathways associated with isobutyrylshikonin in IL-17A-induced HaCaT cell in-flammation.Results:Network pharmacology and molecular docking results indicated that isobutyrylshikonin mainly acts on the JAK/STAT pathway and can spontaneously bind to the key target STAT3 with a binding energy of<-5 kJ/mol.Compared to the control group,isobutyrylshikonin upregulated the mRNA expression levels of CD206,TGF-β,Arg-1,and IL-10.Compared to the model group,isobutyrylshikonin downregulated the mRNA expression levels of TNF-α,iNOS,CXCL9,and CXCL10 in M1 macrophages,reduced the levels of NO and ROS,inhibited the expression of p-JAK2 and p-STAT3 proteins in the JAK/STAT signaling pathway,and suppressed the nuclear translocation of p-STAT3 protein,with statistically significant differences observed(P<0.05).The results of untargeted metabolomics suggested that isobutyrylshikonin may regulate the inflammatory lev-el of IL-17A-induced HaCaT cells through the glutathione metabolism pathway.Conclusion:Isobutyrylshikonin may inhibit LPS-induced inflammation in RAW264.7 cells through the JAK/STAT signaling pathway and regulate the inflammatory of Ha-CaT cells through the glutathione metabolism pathway.关键词
异丁酰紫草素/银屑病/炎症/网络药理学/非靶向代谢组学Key words
Isobutyrylshikonin/Psoriasis/Inflammation/Network pharmacology/Untargeted metabolomics分类
医药卫生引用本文复制引用
MA Yue,MA Yinglan,LI Jianguang,ZHANG Lu..基于网络药理学和非靶向代谢组学探究异丁酰紫草素抗银屑病的作用机制[J].海南医科大学学报,2025,31(23):1823-1832,10.基金项目
This study was supported by the"Tianshan Talent"Young Science and Technology Top-Notch Talent Project(2022TSYCCX0019) （2022TSYCCX0019）
Postdoctoral Science Foundation Project of Xinjiang Medical University(0103010211) （0103010211）
the Third Batch of"Tianchi Talents"Introduction Program of the Autonomous Region-Young Doctoral Talent Project "天山英才"青年科技拔尖人才项目(2022TSYCCX0019) （2022TSYCCX0019）
新疆医科大学博士后科学基金项目(0103010211) （0103010211）
自治区第三批"天池英才"引进计划-青年博士人才项目 （）
