摘要
目的 运用网络药理学探究麻桂柴石方治疗发热的潜在靶点,并通过分子对接和实验验证其解热的作用机制.方法 通过数据库结合文献查找麻桂柴石方中每味药的化合物、潜在靶点及发热的相关靶点,并对靶点进行GO注释、KEGG富集分析和分子对接.通过临床试验和动物实验,验证网络药理学分析结果.结果 筛选出麻桂柴石方化合物265个,潜在靶点435个,与发热的交集靶点110个,涉及到的通路为IL-17、TNF、Toll样受体、NF-κB信号通路等.分子对接显示NOS2、GABRA1、BCL2三个靶点与其相对应的活性成分能结合成稳定构象.临床试验表明,麻桂柴石方治疗外感发热疗效确切.动物实验表明麻桂柴石方能明显降低感染性发热大鼠体温并抑制TLR4、NF-κB p65、COX2、NOS2、GABRA1 mRNA基因表达及TNF-α、IL-1β、IL-6、NOS2、GABRA1血清水平,进一步改善下丘脑组织炎症状态.结论 麻桂柴石方能通过多成分、多靶点、多途径发挥解热作用,实验证实了麻桂柴石方能抑制TLR4/NF-κB/NOS2信号通路mRNA相对表达量,降低SD大鼠炎症因子水平,为深入研究经方合方客观化诊疗提供科学依据.
Abstract
Objective To explore the potential target of Maguichaishi formula in the treatment of fever by using network pharmacology,and to verify its antipyretic mechanism through molecular docking and experiments.Methods The compound components,latent targets and related targets of fever of each drug in Maguichaishi formula were searched through database and literature,and GO annotation,KEGG enrichment analysis and molecular docking were performed on the targets.The rat fever model was established by LPS,and the expression of TLR4/NF-κB/NOS2 signal pathway related genes and pathological changes of hypothalamic tissue with HE staining were observed to verify the results of network pharmacological analysis.Results 265 chemical constituents,435 potential targets and 110 cross targets with fever were screened out from Maguichaishi formula,involving mainly AGE-RAGE,IL-17,TNF,Toll like receptor signal pathway,etc.Molecular docking showed that NOS2,GABRA1,BCL2 and their corresponding active components could combine to form stable conformation.The experiment shows that Maguichaishi formula can significantly reduce the body temperature of febrile rats and inhibit TLR4,NF-κB p65,COX2,NOS2,GABRA1 mRNA gene expression and TNF-α,IL-1β,IL-6,NOS2,GABRA1serum level,further improve the inflammatory state of hypothalamic tissue.Conclusion Maguichaishi formula can play an antipyretic role through multiple components,multiple targets and multiple ways.The experiment confirmed that Maguichaishi formula can inhibit the relative expression of TLR4/NF-κB/NOS2 signal pathway mRNA,reduce the level of inflammatory factors in SD rats,and provide scientific basis for in-depth research on the objective diagnosis and treatment of Jingfang combination.
基金项目
国家中医药局张仲景传承与创新专项(GZY-KJS-2022-047-1)
河南省中医药科学研究专项(2022ZYZD19)
NETL
NSTL
万方数据