Cirsii Herba glycoprotein promotes macrophage M1 polarization through MAPK and NF-κB signaling pathways via interaction with TLR4

Ming Zhao; Shiyue Qin; Jiaqi Wang; Sichun Zheng; Xiaodan Ma; Wei Xu

doi:10.1016/j.ijbiomac.2025.139687

Cirsii Herba glycoprotein promotes macrophage M1 polarization through MAPK and NF-κB signaling pathways via interaction with TLR4

Int J Biol Macromol. 2025 Jan 8:139687. doi: 10.1016/j.ijbiomac.2025.139687. Online ahead of print.

Authors

Ming Zhao¹, Shiyue Qin², Jiaqi Wang³, Sichun Zheng⁴, Xiaodan Ma⁵, Wei Xu⁶

Affiliations

¹ MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
² MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
³ MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
⁴ MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
⁵ Department of Microbiology, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region of China. Electronic address: [email protected].
⁶ MOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].

PMID: 39793782
DOI: 10.1016/j.ijbiomac.2025.139687

Abstract

The present study aimed to extract and purify the glycoprotein from Cirsii Herba (CHPs), and investigate its immunomodulatory activity and molecular mechanism in RAW264.7 macrophages. The results showed that CHPs contained 14.8 % carbohydrates and 80.4 % proteins. CHPs were identified as a glycoprotein around 70 kDa and contained 17 different amino acids, in which the Glu and Asp were predominant. The carbohydrate chain in CHPs was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose and arabinose with the molecular ratio of 6.387: 24.358: 5.766: 8.877: 12.098: 20.427: 7.090: 14.997. CHPs significantly boosted pinocytic and phagocytic activities, increased the secretions of inflammatory factors (NO, TNF-α and IL-6) and chemokines (CXCL2 and CXCL10), and promoted the expressions of accessory and costimulatory molecules (CD40, CD80, CD86, MHC I and MHC II). RNA-seq analysis identified 721 DEGs, 1575 GO terms and 69 KEGG pathways. The pathway inhibition assay presented that MAPK and NF-κB pathways were essential to macrophage activation by CHPs. TLR4 was revealed as a functional receptor and involved in the early recognition of CHPs. These results indicated that CHPs as a glycoprotein promoted macrophage polarization to M1 phenotype mainly via TLR4-dependent MAPK and NF-κB pathways.

Keywords: Cirsii Herba glycoprotein; Immunomodulation; Macrophage; Signaling pathway; TLR4.