Abstract: 【Objective】 The spectrum-effect correlation analysis of the anti-inflammatory and bacteriostatic properties of Lianpu Shuangqing powder was constructed based on experimental design,and its pharmacological substance basis and mechanism of action were revealed through the integration of network pharmacology and in vitro and in vivo experiments.【Method】 The L₉(3⁴) orthogonal experiment was devised incorporating the proportion of Coptidis Rhizoma and Taraxaci herba,the extracted acid concentration and the ethanol concentration,resulting in nine pairs of extracts of Lianpu Shuangqing powder.HPLC was utilized to establish fingerprints for each extract.An in vitro pharmacodynamic analysis of anti-inflammatory and bacteriostatic effects was conducted utilising lipopolysaccharide (LPS)-induced inflammatory cytokines of mouse macrophages and bacteriostasis zone as pharmacodynamic evaluation indexes.The spectrum-effect relationship was established using the grey relational degree analysis and partial least squares regression analysis methods,which screened out the pharmacological substances responsible for its anti-inflammatory effects and bacteriostatic effects against Staphylococcus aureus.Pharmacodynamic constituents and targets were further identified through network pharmacology analysis.Lastly, in vivo experiment was used to verify the spectral and network pharmacology results.【Result】 HPLC fingerprints identified nine common peaks and four components (chicoric acid,coptisine,epiberberine and palmatine).The results of orthogonal test showed that the components in Coptidis Rhizoma played a dominant role in the antibacterial effect against Staphylococcus aureus,while the role of Taraxaci herba in the anti-inflammatory efficacy was more obvious.Spectrum-effect analysis screened that epiberberine and coptisine were part of the pharmacological substance basis for the anti-inflammatory and antibacterial effects of the powder.Network pharmacology research revealed that epiberberine,palmatine,coptisine,luteolin and caffeic acid were the five components with the highest weights in the network.Interleukin-6 (IL6) and tumor necrosis factor (TNF) were the two main anti-inflammatory and antibacterial targets.The docking results showed that the binding energies of five compounds to both target protein receptors were all below －5 kJ/mol,indicating a good binding affinity.In the mouse model of acute enteritis,histological sections revealed that the area of inflammatory cell infiltration in the colon of mice was significantly reduced after treatment with Lianpu Shuangqing powder.ELISA experiments showed a significant difference in the expression of inflammatory factors in colon of mice between the drug group and the model group (P＜0.05).【Conclusion】 An integrated approach involving spectrum-effect correlation analysis based on experimental design and network pharmacology had effectively delineated that the primary pharmacological substances responsible for the anti-inflammatory effects and bacteriostatic effects against Staphylococcus aureus in Lianpu Shuangqing powder were coptisine and epiberberine,with IL6 and TNF being the major target molecules.This study offered a significant exploration into the pharmacological substance basis and target actions of compound traditional Chinese medicines.

Key words: Lianpu Shuangqing powder; experimental design; spectrum-effect relationship; anti-inflammatory and bacteriostatic; pharmacodynamic substance