Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
권호 표지
DOI QR코드

DOI QR Code

Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts anti-inflammatory effect by direct inhibiting toll like receptor 4 signaling pathway

  • Xu, Hong-Lin (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Chen, Guang-Hong (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Wu, Yu-Ting (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Xie, Ling-Peng (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Tan, Zhang-Bin (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Liu, Bin (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Fan, Hui-Jie (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Chen, Hong-Mei (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University) ;
  • Huang, Gui-Qiong (Department of Internal Medicine, Huizhou Hospital of Guangzhou University of Traditional Chinese Medicine) ;
  • Liu, Min (Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University) ;
  • Zhou, Ying-Chun (School of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Nanfang Hospital (ZengCheng Branch), Southern Medical University)
  • Received : 2020.09.27
  • Accepted : 2021.05.24
  • Published : 2022.01.01
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Panax ginseng Meyer (P. ginseng), a herb distributed in Korea, China and Japan, exerts benefits on diverse inflammatory conditions. However, the underlying mechanism and active ingredients remains largely unclear. Herein, we aimed to explore the active ingredients of P. ginseng against inflammation and elucidate underlying mechanisms. Methods: Inflammation model was constructed by lipopolysaccharide (LPS) in C57BL/6 mice and RAW264.7 macrophages. Molecular docking, molecular dynamics, surface plasmon resonance imaging (SPRi) and immunofluorescence were utilized to predict active component. Results: P. ginseng significantly inhibited LPS-induced lung injury and the expression of proinflammatory factors, including TNF-α, IL-6 and IL-1β. Additionally, P. ginseng blocked fluorescencelabeled LPS (LPS488) binding to the membranes of RAW264.7 macrophages, the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs). Furthermore, molecular docking demonstrated that ginsenoside Ro (GRo) docked into the LPS binding site of toll like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) complex. Molecular dynamic simulations showed that the MD2-GRo binding conformation was stable. SPRi demonstrated an excellent interaction between TLR4/ MD2 complex and GRo (KD value of 1.16 × 10-9 M). GRo significantly inhibited LPS488 binding to cell membranes. Further studies showed that GRo markedly suppressed LPS-triggered lung injury, the transcription and secretion levels of TNF-α, IL-6 and IL-1β. Moreover, the phosphorylation of NF-κB and MAPKs as well as the p65 subunit nuclear translocation were inhibited by GRo dose-dependently. Conclusion: Our results suggest that GRo exerts anti-inflammation actions by direct inhibition of TLR4 signaling pathway.

Keywords

Acknowledgement

This study was supported by the National Natural Science Foundation of China (Nos. 81973645, 81673805, 81704058, and 81774100), Natural Science Foundation of Guangdong Province (Nos. 2019A1515011560, 2019B1515120026, and 2019A1515110367), Traditional Chinese Medicine Bureau of Guangdong Province (Nos. 20173016, 20201392), Project of Huizhou science and Technology Bureau (No. 2016C0408023).

References

  1. Takeuchi O, Akira S. Pattern recognition receptors and inflammation. Cell 2010;140(6):805-20. https://doi.org/10.1016/j.cell.2010.01.022
  2. Xiao TS. Innate immunity and inflammation. Cell Mol Immunol 2017;14(1):1-3. https://doi.org/10.1038/cmi.2016.45
  3. Yun M, Yi Y. Regulatory roles of ginseng on inflammatory caspases, executioners of inflammasome activation. J Ginseng Res 2020;44(3):373-85. https://doi.org/10.1016/j.jgr.2019.12.006
  4. Ayaz F. Heteroleptic ruthenium polypyridyl complex had differential effects on the production of pro-inflammatory cytokines TNFα, IL1β, and IL6 by the mammalian macrophages in vitro. Inflammation 2019;42(4):1383-8. https://doi.org/10.1007/s10753-019-00999-y
  5. Yi Y. Ameliorative effects of ginseng and ginsenosides on rheumatic diseases. J Ginseng Res 2019;43(3):335-41. https://doi.org/10.1016/j.jgr.2018.04.004
  6. Hansson GK. Inflammation, atherosclerosis, and coronary artery disease. N Engl J Med 2005;352(16):1685-95. https://doi.org/10.1056/NEJMra043430
  7. Vasefi M, Hudson M, Ghaboolian-Zare E. Diet associated with inflammation and alzheimer's disease. J Alzheimers Dis Rep 2019;3(1):299-309. https://doi.org/10.3233/ADR-190152
  8. Rorato R, Borges BC, Uchoa ET, Antunes-Rodrigues J, Elias CF, Elias L. LPS-induced low-grade inflammation increases hypothalamic JNK expression and causes central insulin resistance irrespective of body weight changes. Int J Mol Sci 2017;18(7):1431. https://doi.org/10.3390/ijms18071431
  9. Schottenfeld D, Beebe-Dimmer J. Chronic inflammation: a common and important factor in the pathogenesis of neoplasia. CA Cancer J Clin 2006;56(2):69-83. https://doi.org/10.3322/canjclin.56.2.69
  10. Rubenfeld GD, Caldwell E, Peabody E, Weaver J, Martin DP, Neff M, Stern EJ, Hudson LD. Incidence and outcomes of acute lung injury. N Engl J Med 2005;353(16):1685-93. https://doi.org/10.1056/NEJMoa050333
  11. Zhao J, Yu H, Liu Y, Gibson SA, Yan Z, Xu X, Gaggar A, Li P, Li C, Wei S, et al. Protective effect of suppressing STAT3 activity in LPS-induced acute lung injury. American Journal of Physiology. Am J Physiol Lung Cell Mol Physiol 2016;311(5):L868-80. https://doi.org/10.1152/ajplung.00281.2016
  12. Ware LB, Matthay MA. The acute respiratory distress syndrome. N Engl J Med 2000;342(18):1334-49. https://doi.org/10.1056/NEJM200005043421806
  13. Bekeredjian-Ding I, Jego G. Toll-like receptors-sentries in the B-cell response. Immunology 2009;128(3):311-23. https://doi.org/10.1111/j.1365-2567.2009.03173.x
  14. Ryu JK, Kim SJ, Rah SH, Kang JI, Jung HE, Lee D, Lee HK, Lee JO, Park BS, Yoon TY, et al. Reconstruction of LPS transfer cascade reveals structural determinants within LBP, CD14, and TLR4-MD2 for efficient LPS recognition and transfer. Immunity 2017;46(1):38-50. https://doi.org/10.1016/j.immuni.2016.11.007
  15. Plociennikowska A, Hromada-Judycka A, Borzecka K, Kwiatkowska K. Cooperation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling. Cell Mol Life Sci 2015;72(3):557-81. https://doi.org/10.1007/s00018-014-1762-5
  16. Billod JM, Lacetera A, Guzman-Caldentey J, Martin-Santamaria S. Computational approaches to toll-like receptor 4 modulation. Molecules 2016;21(8):994. https://doi.org/10.3390/molecules21080994
  17. Iqbal H, Rhee D. Ginseng alleviates microbial infections of the respiratory tract: a review. J Ginseng Res 2020;44(2):194-204. https://doi.org/10.1016/j.jgr.2019.12.001
  18. Wang C, Anderson S, Du W, He T, Yuan C. Red ginseng and cancer treatment. Chin J Nat Med 2016;14(1):7-16. https://doi.org/10.3724/SP.J.1009.2016.00007
  19. Kim J. Pharmacological and medical applications of Panax ginseng and ginsenosides: a review for use in cardiovascular diseases. J Ginseng Res 2018;42(3):264-9. https://doi.org/10.1016/j.jgr.2017.10.004
  20. Yi Y. Ameliorative effects of ginseng and ginsenosides on rheumatic diseases. J Ginseng Res 2019;43(3):335-41. https://doi.org/10.1016/j.jgr.2018.04.004
  21. Lee JH, Min DS, Lee CW, Song KH, Kim YS, Kim HP. Ginsenosides from Korean Red Ginseng ameliorate lung inflammatory responses: inhibition of the MAPKs/NF-κB/c-Fos pathways. J Ginseng Res 2018;42(4):476-84. https://doi.org/10.1016/j.jgr.2017.05.005
  22. Zhu W, Cao FS, Feng J, Chen HW, Wan JR, Lu Q, Wang J. NLRP3 inflammasome activation contributes to long-term behavioral alterations in mice injected with lipopolysaccharide. Neuroscience 2017;343:77-84. https://doi.org/10.1016/j.neuroscience.2016.11.037
  23. Wu YT, Chen L, Tan ZB, Fan HJ, Xie LP, Zhang WT, Chen HM, Li J, Liu B, Zhou YC. Luteolin inhibits vascular smooth muscle cell proliferation and migration by inhibiting TGFBR1 signaling. Front Pharmacol 2018;9:1059. https://doi.org/10.3389/fphar.2018.01059
  24. Gao H, Xiao D, Gao L, Li X. MicroRNA-93 contributes to the suppression of lung inflammatory responses in LPS-induced acute lung injury in mice via the TLR4/MyD88/NF-kB signaling pathway. Int J Mol Med 2020;46(2):561-70. https://doi.org/10.3892/ijmm.2020.4610
  25. Chen L, Su ZZ, Huang L, Xia FN, Qi H, Xie LJ, Xiao S, Chen QF. The AMP-activated protein kinase KIN10 is involved in the regulation of autophagy in arabidopsis. Front Plant Sci 2017;8:1201. https://doi.org/10.3389/fpls.2017.01201
  26. Fan HJ, Zhao XS, Tan ZB, Liu B, Xu HL, Wu YT, Xie LP, Bi YM, Lai YG, Liang HF, et al. Effects and mechanism of action of Huang-Lian-Jie-Du-Tang in atopic dermatitis-like skin dysfunction in vivo and in vitro. J Ethnopharmacol 2019;240:111937. https://doi.org/10.1016/j.jep.2019.111937
  27. Wang Y, Wang C, Cheng Z, Zhang D, Li S, Song L, Zhou W, Yang M, Wang Z, Zheng Z, et al. SPRi determination of inter-peptide interaction by using 3D supramolecular co-assembly polyrotaxane film. Biosens Bioelectron 2015;66:338-44. https://doi.org/10.1016/j.bios.2014.11.025
  28. Li MY, Sun L, Niu XT, Chen XM, Tian JX, Kong YD, Wang GQ. Astaxanthin protects lipopolysaccharide-induced inflammatory response in Channa argus through inhibiting NF-kappaB and MAPKs signaling pathways. Fish Shellfish Immunol 2019;86:280-6. https://doi.org/10.1016/j.fsi.2018.11.011
  29. Zandi E, Rothwarf DM, Delhase M, Hayakawa M, Karin M. The IkappaB kinase complex (IKK) contains two kinase subunits, IKKalpha and IKKbeta, necessary for IkappaB phosphorylation and NF-kappaB activation. Cell 1997;91(2):243-52. https://doi.org/10.1016/S0092-8674(00)80406-7
  30. Cargnello M, Roux PP. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev 2011;75(1):50-83. https://doi.org/10.1128/MMBR.00031-10
  31. Zhang XH, Xu XX, Xu T. Ginsenoside Ro suppresses interleukin-1β-induced apoptosis and inflammation in rat chondrocytes by inhibiting NF-κB. Chin J Nat Med 2015;13(4):283-9. https://doi.org/10.1016/S1875-5364(15)30015-7
  32. Kim S, Oh MH, Kim BS, Kim WI, Cho HS, Park BY, Park C, Shin GW, Kwon J. Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells. J Ginseng Res 2015;39(4):365-70. https://doi.org/10.1016/j.jgr.2015.03.008
  33. Park SB, Park GH, Um Y, Kim HN, Song HM, Kim N, Kim H, Jeong JB. Wood-cultivated ginseng exerts anti-inflammatory effect in LPS-stimulated RAW264.7 cells. Int J Biol Macromol 2018;116:327-34. https://doi.org/10.1016/j.ijbiomac.2018.05.039
  34. Plociennikowska A, Hromada-Judycka A, Borzecka K, Kwiatkowska K. Cooperation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling. Cell Mol Life Sci 2015;72(3):557-81. https://doi.org/10.1007/s00018-014-1762-5
  35. Li MY, Sun L, Niu XT, Chen XM, Tian JX, Kong YD, Wang GQ. Astaxanthin protects lipopolysaccharide-induced inflammatory response in Channa argus through inhibiting NF-κB and MAPKs signaling pathways. Fish Shellfish Immunol 2019;86:280-6. https://doi.org/10.1016/j.fsi.2018.11.011
  36. Brasier AR. The NF-kappaB regulatory network. Cardiovasc Toxicol 2006;6(2):111-30. https://doi.org/10.1385/CT:6:2:111
  37. Gilmore TD. Introduction to NF-kappaB: players, pathways, perspectives. Oncogene 2006;25(51):6680-4. https://doi.org/10.1038/sj.onc.1209954
  38. Li S, Jang H, Zhang J, Nussinov R. Raf-1 cysteine-rich domain increases the affinity of K-Ras/Raf at the membrane, promoting MAPK signaling. Structure 2018;26(3):513-25. https://doi.org/10.1016/j.str.2018.01.011
  39. Liu HM, Lee TY, Liao JF. GW4064 attenuates lipopolysaccharide-induced hepatic inflammation and apoptosis through inhibition of the Toll-like receptor 4-mediated p38 mitogen-activated protein kinase signaling pathway in mice. Int J Mol Med 2018;41(3):1455-62.
  40. Pinzi L, Rastelli G. Molecular docking: shifting paradigms in drug discovery. Int J Mol Sci 2019;20(18):4331. https://doi.org/10.3390/ijms20184331
  41. Zhao S, Yang M, Zhou W, Zhang B, Cheng Z, Huang J, Zhang M, Wang Z, Wang R, Chen Z, et al. Kinetic and high-throughput profiling of epigenetic interactions by 3D-carbene chip-based surface plasmon resonance imaging technology. Proc Natl Acad Sci USA 2017;114(35):E7245-54.
  42. Scarano S, Scuffi C, Mascini M, Minunni M. Surface plasmon resonance imaging (SPRi)-based sensing: a new approach in signal sampling and management. Biosens Bioelectron 2010;26(4):1380-5. https://doi.org/10.1016/j.bios.2010.07.056