Network Pharmacology Analysis and Experimental Investigation of the Anti-Inflammatory Effects of Houttuynia Cordata,Lithospermum Erythrorhizon,and Isatis Indigotica

Yue Zipeng; HE Yinghui; HAO Linyao; Huo Shuying; LI Yurong; LI Shupeng

doi:10.19978/j.cnki.xmsy.2025.05.02

Abstract：To investigate the anti-inflammatory effects of the combination of Houttuynia cordata,Rubia cordifolia,and Isatis indigotica on macrophage RAW264.7 cells through network pharmacology,molecular docking,and experimental verification methods.The active components and related target sites of Houttuynia cordata,Rubia cordifolia,and Isatis indigotica were identified using TCMSP,ETCM,UniProt,and SwissTargetPrediction databases;the disease targets of inflammation were obtained using OMIM,TTD,and Genecard databases;the drug-active component-inflammation target network diagram was drawn using Cytoscape 3.6.1 software,and GO function and KEGG pathway enrichment analyses were performed;molecular docking was conducted using AutoDock software.The mRNA secretion of TNF-α,IL1-β,and IL-6 in macrophage RAW264.7 cells was detected by real-time fluorescent quantitative PCR to evaluate the anti-inflammatory effect of the herbal mixture.The anti-inflammatory effect of Houttuynia cordata,Rubia cordifolia,and Isatis indigotica combination was detected by mouse ear swelling test.A total of 702 active components were obtained for Houttuynia cordata,Rubia cordifolia,and Isatis indigotica,including scutellarin,corindone,and kaempferol,etc.;675 drug-inflammation intersection targets were obtained,including IL6,IL1β,and serine/threonine protein kinase(AKT1),etc.;GO function enrichment analysis showed 186,36,and 86 entries;KEGG result analysis yielded 189 pathways.Molecular docking results showed that the active ingredients of the drugs had good binding ability with the main targets.In vitro experiments demonstrated that the three herbal medicines could effectively inhibit the caused by Lipopolysaccharide(LPS),with pathological increases of inflammatory factors such as IL-6,TNF-α,and IL-1β.The combination of Houttuynia cordata,Rubia cordifolia,and Isatis indigotica inhibited the ear swelling of mice caused by dexamethasone at 47.61%,which was significantly different from the model group,indicating that the herbal mixture can exert anti-inflammatory effects.The good anti-inflammatory effect of the combination of Houttuynia cordata,Rubia cordifolia,and Isatis indigotica is demonstrated by network pharmacology analysis and experimental results,suggesting that it exerts anti-inflammatory effects through multiple components,multiple pathways,and multiple targets.