Exploring the Mechanism of Duhuojisheng Pill Against Rheumatoid Arthritis by Network Pharmacology and Experimental Validation

Abstract

Ethnopharmacological relevance: DuhuoJisheng pill (DHJS), a classic formula of traditional Chinese medicine (TCM), has been widely used in the treatment of rheumatoid arthritis (RA) in China for many years. However, the effective components and therapeutic mechanisms of DHJS for treating RA are still unclear.

Aim of the study: To elucidate the potential mechanism of DHJS for the treatment of RA based on network pharmacology and experimental validation in vivo.

Materials and methods: The main active ingredients and targets of DHJS for treating RA were screened by searching the online database. Subsequently, the protein-protein interaction (PPI) network of potential targets of DHJS in the treatment of RA was constructed, and the key targets were determined through the topological analysis of PPI. Then, these key targets were enriched by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), and ultimately the potential signaling pathways for DHJS to treat RA were screened. The collagen-induced arthritis model was established, and the results of network pharmacologic analysis of DHJS against RA were verified in vivo.

Results: In all, 33 intersecting targets of DHJS and RA, and 17 key targets were identified by topological analysis of PPIs. The active ingredients of DHJS against RA may be MOL000422 (kaempferol), MOL000173 (wogonin), MOL000449 (stigmasterol) and MOL000098 (quercetin) by herb-compound-target network diagram. Further, GO and KEGG analysis revealed that the 17 targets might be involved in the inflammatory response, immune response, tumor necrosis factor (TNF) and NF- κB and PI3K / Akt pathways. The experimental validation indicated that DHJS treatment could alleviate ankle inflammation and improve bone destruction in CIA rats. DHJS could effectively inhibit the release of inflammatory cytokines including TNF-α, IL-1β and IL-6. Moreover, the protein expression of p-PI3K, p-AKT and p-p65 in spleen tissue decreased after DHJS treatment.

Conclusion: Our discoveries revealed that DHJS significantly inhibited the CIA-induced inflammatory response via PI3K/AKT/NF-κB signaling pathway.

Note:

Funding Information: This study was supported by the Natural Science Foundation of Heilongjiang Province (No. YQ2019H005), the National Natural Science Foundation of China (No. 81903763), the China Postdoctoral Science Foundation (No. 2019M661312), the Fundamental Research Funds for the Provincial Universities (JFWLD201904, No.2018XN-25), the Nanjing medical science and technology development fund (No. YKK20179), and the Initial Scientific Research Fund of the Talents Introduced in Nanjing Lishui People’s Hospital (No. 2021YJ01).

Declaration of Interests: The authors declare no conflict of interest.

Ethical Approval Statement: The experimental procedure was conducted in accordance with NIH guidelines and approved by the experimental animal ethics committee of Harbin Medical University (HMUDQ20220517001).

Keywords: DuhuoJisheng pill, Rheumatoid arthritis, Network Pharmacology, PI3K/AKT/NF-κB pathway