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Identification of intracellular activation mechanism of rhamnogalacturonan-I type polysaccharide purified from Panax ginseng leaves in macrophages and roles of component sugar chains on activity

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Abstract

This study aimed to investigate the mechanisms underlying intracellular signaling pathways in macrophages in relation to the structural features of rhamnogalacturonan (RG) I-type polysaccharide (PGEP-I) purified from Panax ginseng leaves. For this investigation, we used several specific inhibitors and antibodies against mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB), and pattern recognition receptors (PRRs). Furthermore, we investigated the roles of component sugar chains on immunostimulating activity through a sequential enzymatic and chemical degradation steps. We found that PGEP-I effectively induced the phosphorylation of several MAPK- and NF-κB-related proteins, such as p38, cJun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p65. Particularly, immunocytochemistry analysis confirmed the PGEP-I-induced translocation of p65 into the nucleus. Furthermore, the breakdown of PGEP-I side chains and main chain during sequential enzymatic and chemical degradation reduced the PGEP-I-induced macrophage cytokine secretion activity. IL-6, TNF-α, and NO secreted by macrophages are associated with several signaling pathway proteins such as ERK, JNK, and NF-κB and several PRRs such as dectin-1, CD11b, CD14, TLR2, TLR4, and SR. Thus, these findings suggest that PGEP-I exerts potent macrophage-activating effects, which can be attributed to its typical RG-I structure comprising arabinan, type II arabinogalactan, and rhamnose-galacturonic acid repeating units in the main chain.

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Acknowledgements

This study was supported by a 2018 grant from the Korean Society of Ginseng funded by the Korean Ginseng Corporation. This work was supported by Kyonggi University‘s Graduate Research Assistantship 2023.

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  1. Seung-U Son and Hee Won Lee contributed equally to this work.

Authors and Affiliations

  1. Department of Food Science and Biotechnology, Kyonggi University, Suwon, 16227, Republic of Korea

    Seung-U Son, Hee Won Lee, Ju-Hyeon Park & Kwang-Soon Shin

  2. Transdisciplinary Major in Learning Health System, Department of Integrated Biomedical and Life Science, Korea University, Seoul, 02841, Republic of Korea

    Seung-U Son

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SUS: experiments, methodology, writing of the original draft, reviewing, and editing; HWL: experiments, methodology; JHP: experiments; KSS: conceptualization, methodology, and project administration.

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Correspondence to Kwang-Soon Shin.

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Son, SU., Lee, H.W., Park, JH. et al. Identification of intracellular activation mechanism of rhamnogalacturonan-I type polysaccharide purified from Panax ginseng leaves in macrophages and roles of component sugar chains on activity. J Nat Med 78, 328–341 (2024). https://doi.org/10.1007/s11418-023-01768-w

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