| Objective:To establish a network of “active components-targets-diseases” of breviscapine for improving insulin resistance by using network pharmacology approach and define the corresponding biological functions and signal pathways,to explain the anti-insulin resistance of breviscapine scientifically and validate the possible functional target for its quality evaluation.And thus lay the experimental foundation for follow-up research and further clinical application of breviscapine.Methods:The main active components of breviscapine were determined by TCMSP databases and literature mining.Multiple databases were used to search and screen the corresponding targets of active components.The “active components-targets-diseases” interactive network was constructed and its docking verification was conducted through the SystemsDock web site.The targets biological functions and related pathways were analyzed by DAVID combined with ClueGO plugin.Based on the network analysis,we successfully established a model of insulin resistance in HepG2 cells.The effect of breviscapine on cell glucose consumption was determined by using the glucose oxidase(god-pod)method.The cell morphology and intracellular lipid drip formation were observed by Oil red O staining.The cell viability was detected by cell counting kit-8(CCK-8).The expression changes of PTP1 B,AKT and p-AKT in insulin-resistant HepG2 cells treated with breviscapine were observed by qRT-PCR and Western blot.Results:Verified by the database retrieval and literature,scutellarin was reported to be the main active component of breviscapine.Network topology analysis identified four potential key targets closely related to insulin resistance therapy(including NOS3,PTPN1,ADORA1,and PRKCG),which participated in biological processes such as metabolic regulation,inflammatory reaction,and apoptosis regulation.Anti-insulin resistance was exerted by intervention of related signaling pathways such as PI3K-Akt,Ras,insulin resistance and HIF-1.The vitro experimental results showed that breviscapine could increase glucose uptake and utilization in HepG2 cells insulin resistance model,down-regulate the expression of PTP1 B mRNA and protein,and effectively increase the phosphorylation level of AKT.Conclusion:Breviscapine improving insulin resistance is a multi-target,multi-path,and multichoice process with complex mechanism,which provides a basis for in-depth research on its mechanism of action.In vitro validation of PTP1 B,the main target of breviscapine for insulin resistance suggests that the improvement of insulin resistance by breviscapine may be related to the decreased expression of PTP1 B and the promotion of AKT activation. |
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