Unveiling the immunomodulatory mechanism of polysaccharides from Polygonum cyrtonema based on RNA-seq

• Published in: Food research international Vol. 175; p. 113755
• Main Authors: Shan, Jialuo, Ma, Wenjie, Guo, Yue, Chang, Xinxin, Xie, Jianhua, Chen, Yi, Hu, Xiaobo, Yu, Qiang
• Format: Journal Article
• Language: English
• Published: Canada 01.01.2024
• Subjects: Immunity; NF-kappa B - genetics; NF-kappa B - metabolism; Polygonum - metabolism; Polysaccharides - pharmacology; RNA-Seq; Tumor Necrosis Factor-alpha; NF-κB; Polygonatum cyrtonema polysaccharide; RNA-seq; JAK-STAT; MAPKs
• ISSN: 0963-9969; 1873-7145
• DOI: 10.1016/j.foodres.2023.113755

This work aimed to illuminate the mechanism of Polygonatum cyrtonema polysaccharide (PCP-80%) triggered immune activation. Results showed that PCP-80% enhanced the protein expression of COX-2 and iNOS, along with increasing the release of NO, ROS, cytokines (TNF-α, IL-6) in RAW264.7 cells. RNA-seq analysis revealed 2160 differentially expressed genes (DEGs) following PCP-80% treatment, comprising 1142 up-regulated and 1018 down-regulated genes. In addition, for investigating possible regulatory mechanisms, the NF-κB, MAPKs, and JAK-STAT signaling pathways were also chosen based on bioinformatics analysis. Furthermore, these findings were further corroborated through Western blot experiments, validating the activation of JAK-STAT (reduction of JAK1 in cells and elevation of p-STAT3 in the nucleus), MAPK (elevation of p-p38, p-ERK1/2, and p-JNK), and NF-κB (elevation of p-IκBα in cells, reduction of cytoplasmic p65, and increase of nuclear content of p-p65) in macrophage activation induced by PCP-80%. Besides, the production of NO and TNF-α was decreased by the inhibitor of the three pathways. In conclusion, these findings provide strong evidence that PCP-80% effectively modulates the immune response of macrophages, with significant involvement of the JAK-STAT, MAPKs, and NF-κB signaling pathways.