Title:Integrating Network Pharmacology and Experimental Validation to
Decipher the Anti-Inflammatory Effects of Magnolol on LPS-induced
RAW264.7 Cells
Volume: 27
Issue: 3
Author(s): Lei Hao, Xiaoying Zhong, Runjia Yu, Jiahui Chen, Wei Li, Yuzhong Chen, Weiqi Lu, Jianyu Wu and Peizong Wang*
Affiliation:
- State Key Laboratory of Oncology in South
China, Department of Anesthesiology, Sun Yat-Sen University Cancer Center, Collaborative Innovation Center for
Cancer Medicine, Guangzhou, Guangdong, 510060, P.R. China
Keywords:
Magnolol, anti-inflammatory effects, macrophages, RAW264.7 cells, PI3K-Akt, network pharmacology.
Abstract:
Introduction: Magnolol is beneficial against inflammation-mediated damage. However,
the underlying mechanisms by which magnolol exerts anti-inflammatory effects on macrophages
remain unclear.
Objective: In this study, network pharmacology and experimental validation were used to assess
the effect of magnolol on inflammation caused by lipopolysaccharide (LPS) in RAW264.7 cells.
Materials and Methods: Genes related to magnolol were identified in the PubChem and Swiss
Target Prediction databases, and gene information about macrophage polarization was retrieved
from the GeneCards, OMIM, and PharmGKB databases. Analysis of protein-protein interactions
was performed with STRING, and Cytoscape was used to construct a component-target-disease
network. GO and KEGG enrichment analyses were performed to ascertain significant molecular
biological processes and signaling pathways. LPS was used to construct the inflammatory cell
model. ELISA and qRTPCR were used to examine the expression levels of inflammationassociated
factors, immunofluorescence was used to examine macrophage markers (CD86 and
CD206), and western blotting was used to examine protein expression levels.
Results: The hub target genes of magnolol that act on macrophage polarization were MDM2,
MMP9, IL-6, TNF, EGFR, AKT1, and ERBB2. The experimental validation results showed that
magnolol treatment decreased the levels of proinflammatory factors (TNF-α, IL-1β, and IL-6).
Moreover, the levels of anti-inflammatory factors (IL-10 and IL-4) were increased. In addition,
magnolol upregulated the expression of M2 markers (Agr-1, Fizzl, and CD206) and downregulated
M1 markers (CD86). The cell experiment results supported the network pharmacological
results and demonstrated that magnolol alleviated inflammation by modulating the PI3k-Akt and
P62/keap1/Nrf2 signaling pathways.
Conclusion: According to network pharmacology and experimental validation, magnolol attenuated
inflammation in LPS-induced RAW264.7 cells mainly by inhibiting M1 polarization and
enhancing M2 polarization by activating the PI3K/Akt and P62/keap1/Nrf2 signaling pathways.
