인삼의 선천면역 조절을 위한 인삼의 있어서 면역조절 역할 = Immunemodulative Roles of Panax ginseng in regulating Innate Immunity

다국어 초록 (Multilingual Abstract)

Korean Red Ginseng (KRG), a heat-processed form of Panax ginseng, is widely recognized for its immunomodulatory properties, primarily attributed to its bioactive ginsenosides. Although extensive studies have examined its immune-enhancing potential in humans and rodents, limited information exists regarding its effects in pig models that more closely resemble humans in terms of immunology. This study investigated the immunomodulatory effects of KRG in porcine models by evaluating immune cell activity, cytotoxic responses, cytokine production, and gene expression profiles. Twenty-four male Yorkshire piglets were orally administered KRG at 3 g/day or 6 g/day for 14 days. In vitro assays revealed that KRG treatment (100 and 200 µg/mL) did not induce cytotoxicity in splenocytes but significantly enhanced their cytotoxic activity against Yac-1 target cells at higher effector-to-target ratios. Flow cytometry demonstrated significant increases in CD4⁺, CD8⁺, CD4⁺CD8⁺ T cells, NK cells, and CD172⁺ monocytes following KRG administration. Molecular analysis revealed the upregulation of granzyme B, NKp30, and NKp46 mRNA, with NKp44 expression being explicitly elevated in the higher-dose group, indicating a dose-dependent enhancement of cytotoxic pathways. Serum cytokine measurements further revealed significant increases in IL-2, IL-4, IL-6, and IL-10, suggesting a balanced activation of both Th1 and Th2 responses.
Collectively, these findings demonstrate that KRG enhances innate and adaptive immune functions in porcine models at cellular, molecular, and functional levels. This study supports the potential of KRG as an immunomodulatory supplement in veterinary applications. It provides a translational foundation for future studies exploring its therapeutic or prophylactic use in both animal and human health.
Gintonin (GT), a bioactive component of Panax ginseng, exhibits significant immunomodulatory properties, affecting macrophage activity, Natural Killer cell cytotoxicity, and the expression of key inflammatory mediators and signaling pathways. This study demonstrates that GT enhances the viability of RAW 264.7 macrophages in a concentration-dependent manner, concurrently inducing a dose-dependent increase in nitric oxide production, indicating macrophage activation. Furthermore, GT treatment in vivo upregulated crucial surface activation markers—CD11b, CD80, and MHC class II—on peritoneal macrophages, suggesting improved adhesion, co-stimulation, and antigen presentation. GT also significantly boosted NK cell-mediated cytotoxicity, as evidenced by a dose-dependent increase in K562 cell apoptosis, indicating enhanced immune surveillance. At the molecular level, GT profoundly influenced inflammatory signaling. It increased the mRNA expression of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β, as well as components of the inflammasome (iNOS, NLRP3, caspase-1).
GT also activated critical signaling pathways, as demonstrated by increased phosphorylation of JNK and p38 in the MAPK pathway, and upregulation of NF-κB and its downstream target, p65. A notable finding is GT's dual immunomodulatory role. While it enhanced pro-inflammatory markers, it simultaneously raised the anti-inflammatory cytokine TGF-β and significantly reduced COX-2 expression. This balanced action suggests that GT not only promotes immune activation but also helps to mitigate excessive inflammation. Collectively, these findings underscore GT's potential as a sophisticated immunomodulator that can both enhance immune responses and maintain inflammatory homeostasis, positioning it as a promising natural agent for therapeutic applications.
Polysaccharide (PS) is a fraction of Panax ginseng that has several features that benefit health. In this study, we investigated the immune-enhancing effect of PS, focusing on the role of PBMCs and spleen-mediated immune functions. Our aim to reveal the mechanisms underlying Ginseng PS-mediated regulation of immune cells using the cyclophosphamide (CPA) 30 mg/kg induced immunosuppressed rat model. In vivo investigation revealed that ginseng PS administration can increase the tissue weight of major immune organs, such as the spleen and thymus. PS reversed CPA-induced immunosuppression and increased the number of immune cells, such as CD3⁺, CD4⁺T, CD8⁺, NK cells (CD161⁺, NkG2D), also activated macrophage Markers CD11b/c⁺, and MHC-II in the PBMCs and spleen. Moreover, ginseng PS alleviated immunosuppression by up-regulating TLR receptors, regulating MAPKs and nuclear factor Kappa-B(NF-κB) p50/p65 pathways, by increasing secretion of cytokines, including TNF-α, IFN-γ, IL-1β, interleukin (IL)-2, IL-6, and IL-10. PS shows great potential as an immune enhancer and a natural medicine for treating the immunosuppression model. Overall, our study demonstrates the immune-enhancing effect of PS, suggesting its potential as a medicinal or therapeutic agent for pathological conditions involving immunosuppression.

목차 (Table of Contents)

• CHAPTER I 1
• Abstract(ENGLISH) 1
• 1. Introduction 3
• 2. Material &Method 9
• 2.1 Chemical and Reagent 9
• 2.2 Animals 10
• 2.3 PBMC isolation 10
• 2.4 Cytotoxicity and Functional Assays of Porcine Splenocytes 11
• 2.5 Flow cytometry analysis 12
• 2.6 MACS separation step 13
• 2.7 PCR (band) 13
• 2.8 Serum Cytokine Quantification 14
• 2.9 Statistical analysis 14
• 3. Results 15
• 3.1 In vitro, Effects of KRG on Splenocytes viability and Cytotoxic Function 15
• 3.2 NK cell purification 15
• 3.3 Effects of KRG Immune Cell population by Flow cytometry 16
• 3.4 KRG Enhances mRNA Expression of Cytotoxicity Markers. 16
• 3.5 KRG Enhances Cytokine Production 17
• 4. Discussion 17
• 5. Conclusion 21
• 6. References 22
• 7.Appendix 1 29
• CHAPTER II 48
• Abstract(ENGLISH) 48
• 1. Introduction 50
• 2. Material & Method 54
• 2.1 Chemical and Reagent 54
• 2.2 Cell Culture 54
• 2.3 Design for experiment 55
• 2.4 Assessment of Cell Viability 56
• 2.5 NO production 56
• 2.6 Cytotoxicity analysis 57
• 2.7 Flow cytometric analysis 57
• 2.8 RT-PCR analysis 58
• 2.9 Western Blotting 58
• 2.10 Statistical analysis 59
• 3. Results 59
• 3.1 Effects of Gintonin on Cell Viability and Nitric Oxide Production in RAW 264.7 Macrophages 59
• 3.2 Effect of Gintonin on NK Cell Cytotoxicity and Macrophage Activation 60
• 3.3 Immunomodulatory Effects of Gintonin on Pro-Inflammatory Signaling and Immune-Related Gene Expressio 61
• 3.4 Effect of Gintonin on Cytokine Production and Immune-Related Protein Expression Analyzed by Western Blot 62
• 4. Discussion 64
• 5. Conclusion 67
• 6. References 68
• 7.Appendix 2 75
• CHAPTER III 103
• Abstract(ENGLISH) 103
• 1.Introduction 105
• 2 Material &Method 109
• 2.1 Chemical and Reagent 109
• 2.2 Cell Culture 110
• 2.3 Experiment design 110
• 2.4 Cell Viability Assessment Following Ginseng PS Treatment 111
• 2.5 Peripheral Blood Mononuclear Cell (PBMC) Isolation 111
• 2.6 Splenocytes Isolation 112
• 2.7 Flow Cytometry Analysis of Cell Surface Markers 112
• 2.8 RNA Extraction and Gene Expression Analysis 113
• 2.9 Statistical analysis 114
• 3. Results 114
• 3.1 In vitro, The Effect of Ginseng PS on the viability of RAW 264.7 cells 114
• 3.2 Ginseng PS enhances the expression of NK Cell activation markers in PBMCs and Splenocytes 115
• 3.3 Ginseng PS Promotes Activation of CD4 and CD8 T Cells 115
• 3.4 Ginseng PS Enhances Activation of CD11b/c and MHC-II Cells 116
• 3.5 Ginseng PS Enhances Activation of Granulocytes 116
• 3.6 Ginseng PS Modulates mRNA Expression of Immune Cytokines in PBMCs and SP 117
• 3.7 Effects of Ginseng PS Treatment on mRNA Expression of Cytotoxic Receptor Genes in PBMCs and SP 118
• 3.8 Effects of Ginseng PS Treatment on mRNA Expression of TLRs and MAPK Pathway Genes in PBMCs and Splenocytes 118
• 3.9 Effects of Ginseng PS on the Expression of NF-κB Pathway Components and COX-2 in PBMCs and Splenocytes 120
• 4. Discussion 121
• 5. Conclusion 126
• 6. References 127
• Appendix 3 133
• 7. Abstract(Korean) 187