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  2. Peucedanum praeruptorum Dunn polysaccharides regulate macrophage inflammatory response through TLR2/TLR4-mediated MAPK and NF-κB pathways

Peucedanum praeruptorum Dunn polysaccharides regulate macrophage inflammatory response through TLR2/TLR4-mediated MAPK and NF-κB pathways

  • Biomed Pharmacother. 2022 Aug;152:113258. doi: 10.1016/j.biopha.2022.113258.
Ming Zhao 1 Jing Hou 2 Sichun Zheng 3 Xiaodan Ma 4 Xinyu Fu 5 Songhua Hu 6 Kai Zhao 7 Wei Xu 8
Affiliations

Affiliations

  • 1 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 2 Instrumental Analysis Center of Zhejiang Gongshang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 3 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 4 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China; Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 5 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 6 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
  • 7 Institute of Nanobiomaterials and Immunology, School of Life Science, Taizhou University, Taizhou 318000, China. Electronic address: [email protected].
  • 8 Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China; Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
Abstract

The present study was to investigate the molecular mechanisms underlying macrophage inflammatory response to Polysaccharides from Peucedanum praeruptorum Dunn (PPDs) and elucidate the receptors and signaling pathways associated with PPDs-mediated macrophage activation. MTT and Griess method were performed to investigate the effects of PPDs on cell viability and NO production. Neutral red and FITC-dextran were used to determine the pinocytic and phagocytic activity. RT-qPCR and ELISA were employed to analyze the mRNA expression of inflammatory factors and production of cytokines and chemokines. RNA-seq and bioinformatics analysis were conducted to determine the underlying molecules, regulators and pathways, which were further validated by pathway inhibition and neutralization assays. The results indicated that PPDs significantly enhanced pinocytic and phagocytic activity, promoted the expression and secretion of inflammatory factors and chemokines, and boosted the expression of accessory and costimulatory molecules. RNA-Seq analysis identified 1343 DEGs, 405 GO terms and 91 KEGG pathways. IL6 and TNF were identified as hubs of connectivity in PPDs-mediated macrophage activation. "Cytokine-cytokine receptor interaction", "TNF signaling pathway", "NF-kappa B signaling pathway", "JAK-STAT signaling pathway" and "MAPK signaling pathway" were the most significant pathways. The pathway inhibition assay revealed that MAPK and NF-κB pathways were essential to macrophage activation by PPDs. TLR2 and TLR4 were uncovered to be the functional receptors and involved in recognition of PPDs. These results indicated that PPDs modulated macrophage inflammatory response mainly through TLR2/TLR4-dependent MAPK and NF-κB pathways.

Keywords

Inflammatory response; Macrophage; Pathway; Peucedanum praeruptorum Dunn; Polysaccharides; TLR.

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