关键词: 强直性脊柱炎, 附着点炎, 网络药理学, 青风藤, 青藤碱

Abstract:

Objective： To explore the mechanism of Sinomenium acutum in the treatment of enthesitis of ankylosing spondylitis （AS） by using network pharmacology combined with cell experiments.Methods： The active components and potential targets of Sinomenium acutum were screened and summarized through traditional Chinese medicine databases and analysis platforms such as TCMSP/BATMAN. The relevant targets of AS were retrieved in databases such as OpenTargets/DrugBank/GeneCards， and then a compound-target-disease Venn diagram was constructed to obtain the list of intersection targets between drug targets and disease targets. The drug-component-target-disease network diagram was drawn by using Cytoscape 3.9.1 software. The protein-protein interaction （PPI） was obtained through the STRING database to screen the core targets among the intersection targets. The GO function and KEGG pathway enrichment analysis were performed by using the clusterProfiler package of R 4.2.1 software to explore the potential mechanism of Sinomenium acutum on AS， and the drug-component-target-pathway network diagram was constructed by using Cytoscape. The results of network pharmacology analysis were verified by molecular docking. Experimental verification： BALB/c mouse fibroblast-like synoviocytes （FLS） and IL-1β-induced mouse FLS were taken as the research objects and divided into blank control group， IL-1β group， and positive drug group. The key targets predicted by network pharmacology were detected by ELISA and Western blot.Results： Through network pharmacology， 22 active components and 582 targets of Sinomenium acutum were screened out， and 770 disease-related targets were obtained， with 101 common targets. GO function enrichment analysis obtained 1 379 biological process （BP） items， 14 cellular component （CC） items， and 96 molecular function （MF） items. 153 significant signal pathways were enriched by KEGG. Molecular docking showed that the main active components of Sinomenium acutum （kaempferol， stepholidine， sinomenine， crocetin， etc.） had good affinity with core targets such as IL6， TNF， IL1β， PTGS2. In the experiment， sinomenine， the main active component of Sinomenium acutum， was selected to intervene in IL-1β-induced FLS. The results of ELISA and Western blot showed that compared with the IL-1β-induced group， the expressions of TNF-α， PTGS2， Bcl-2， and IL-17 in the positive drug group were decreased， and the expressions of PTGS2 and Bcl-2 were dose-dependent， while Bcl-2 was significantly increased （P0.01）， and there was no significant difference in MMP9 （P0.05）.Conclusion： The main component SIN in Sinomenium acutum may inhibit the inflammatory response and induce apoptosis by down-regulating core targets represented by PTGS2， Bcl-2， IL-17， etc. and regulating pathways， thus acting synergistically on the treatment of AS enthesitis through multiple pathways.

Key words: Ankylosing spondylitis； , Enthesitis of ankylosing spondylitis； , Network pharmacology； , Sinomenium acuturn； , Sinomenine