Study On Effect Of Compatibility Of Scutellariae Radix And Fritillariae Cirrhosae Bulbus On The Main Chemical Components In Scutellariae Radix And Its Anti-pneumonia Effect And Mechanism

Objective: To explore the effects of Compatibility of Scutellariae Radix and Fritillariae Cirrhosae Bulbus in different proportions on the main chemical components in Scutellariae Radix,and to explore the effect and mechanism of its anti-LPS-induced pneumonia,and provide experimental basis for clinical application.Methods: 1.The UPLC-Q Ex Active Orbitrap/MS was used to qualitatively analyze the chemical components in the water extract of Scutellariae Radix and Fritillariae Cirrhosae Bulbus.The content of baicalin,wogonoside,baicalein and wogonin in the water extract of Scutellariae Radix and Fritillariae Cirrhosae Bulbus with the ratio of 3:0,3:1,3:2,3:3,2:3,1:3 and 0:3 were determined by HPLC.2.The 72 mice were divided into blank group,model group,positive group,Scutellariae Radix low,medium and high groups,Fritillariae Cirrhosae Bulbus low,medium and high groups,Scutellariae Radix-Fritillariae Cirrhosae Bulbus low,medium and high groups;The mice in the experimental groups were administered for six consecutive days.After 30 minutes of administration on the seventh day,LPS(10 mg/kg)was injected intraperitoneally for 6 hours.The levels of IL-6,IL1β and TNF-α in serum and lung tissues were determined by ELISA.The pathological changes of lung were observed by HE staining.3.The active components and targets of Scutellariae Radix-Fritillariae Cirrhosae Bulbus were collected by TCMSP,Genecards and Bioconductor databases;The active component-target network construction,GO function and KEGG pathway enrichment analysis and other network pharmacological studies were carried out using String,Cytoscape,R and other software.The molecular docking model was established by Maestro11.9 software to verify the interaction between the active component and the target.4.Alveolar macrophages were cultured and divided into blank group,model group(LPS 1μg/m L),Scutellariae Radix-Fritillariae Cirrhosae Bulbus low-dose,medium-dose and high-dose groups.The expression of inflammatory cytokines including IL-6,IL-1β and TNF-α was detected by ELISA.The protein expression of p-IκBα and IκBα in cells was detected by Western blot.Results: 1.Twenty-one chemical components were identified by UPLC-Q Exactive Orbitrap/MS,including 19 components of Scutellariae Radix and 2 components of Fritillariae Cirrhosae Bulbus.Determination of baicalin,baicalein,wogonin and wogonin in water extracts of different proportions of Scutellariae Radix-Fritillariae Cirrhosae Bulbus by HPLC.When the compatibility ratio of the Scutellariae Radix-Fritillariae Cirrhosae Bulbus is in the range of 3:0～2:3,the content change trend of baicalin and wogonin is similar,the content change trend of baicalin and wogonin is similar,and the total content change rate of the four chemical components is small.When the compatibility ratio reached 1:3,the total content of the four chemical components dropped by 23.7%.2.Before and after the compatibility of Scutellariae Radix and Fritillariae Cirrhosae Bulbus could inhibit the contents of IL-1β,IL-6 and TNF-α inflammatory factors in serum and lung tissue of pneumonia mice,and improve the pathological structure of lung tissue of pneumonia mice.3.In the network pharmacological study,49 active components and 33 active components-disease targets of Scutellariae Radix and Fritillariae Cirrhosae Bulbus were screened out;In PPI network analysis,nodes with Degree≥10 had 18 core targets such as IL-6,accounting for 51.52% of the total.The enrichment analysis revealed 52 GO functions such as nuclear receptor activity and 93 signaling pathways such as IL-17 and NF-κB.Molecular docking studies showed that PTGS1 had strong binding ability with Ent-epicatechin,Norwogoni,5,2’,6’-trihydroxy-7,8-dimethoxyflavone.AR has strong binding ability with Ent-Epicatechin,Baicalein and Oroxylin A.4.The compatibility of Scutellariae Radix and Fritillariae Cirrhosae Bulbus can inhibit cell secretion of IL-6,IL-1β,TNF-α inflammatory factors and IκBα phosphorylation,so that the ratio of p-IκBα/IκBα is reduced,there by inhibiting the activation of NF-κB signaling pathway.Conclusion: The content of baicalin,wogonoside,baicalein and wogonin in Scutellariae Radix were affected by the compatibility of Fritillariae Cirrhosae Bulbus,and the compatibility ratio should be 3:1～2:3.Before and after its equal ratio compatibility,it has protective effect on mice with pneumonia,and the mechanism of action is realized by regulating NF-κB and other signal pathways.