Abstract
The effects of Korean red ginseng extract (KRGE) on autoimmune disorders of the nervous system are not clear. We investigated whether KRGE has a beneficial effect on acute and chronic experimental autoimmune encephalomyelitis (EAE). Pretreatment (daily from 10 days before immunization with myelin basic protein peptide) with KRGE significantly attenuated clinical signs and loss of body weight and was associated with the suppression of spinal demyelination and glial activation in acute EAE rats, while onset treatment (daily after the appearance of clinical symptoms) did not. The suppressive effect of KRGE corresponded to the messenger RNA (mRNA) expression of proinflammatory cytokines (tumor necrosis factor-α [TNF-α] and interleukin [IL]-1β), chemokines (RANTES, monocyte chemotactic protein-1 [MCP-1], and macrophage inflammatory protein-1α [MIP-1α]), adhesion molecules (intercellular adhesion molecule-1 [ICAM-1], vascular cell adhesion molecule-1 [VCAM-1], and platelet endothelial cell adhesion molecule [PECAM-1]), and inducible nitric oxide synthase in the spinal cord after immunization. Interestingly, in acute EAE rats, pretreatment with KRGE significantly reduced the population of CD4+, CD4+/IFN-γ+, and CD4+/IL-17+ T cells in the spinal cord and lymph nodes, corresponding to the downregulation of mRNA expression of IFN-γ, IL-17, and IL-23 in the spinal cord. On the other hand, KRGE pretreatment increased the population of CD4+/Foxp3+ T cells in the spinal cord and lymph nodes of these rats, corresponding to the upregulation of mRNA expression of Foxp3 in the spinal cord. Interestingly, intrathecal pretreatment of rats with ginsenosides (Rg1 and Rb1) significantly decreased behavioral impairment. These results strongly indicate that KRGE has a beneficial effect on the development and progression of EAE by suppressing T helper 1 (Th1) and Th17 T cells and upregulating regulatory T cells. Additionally, pre- and onset treatment with KRGE alleviated neurological impairment of myelin oligodendrocyte glycoprotein35–55-induced mouse model of chronic EAE. These results warrant further investigation of KRGE as preventive or therapeutic strategies for autoimmune disorders, such as multiple sclerosis.
Similar content being viewed by others
Abbreviations
- APG:
-
Acidic polysaccharide of Panax ginseng
- BBB:
-
Blood–brain barrier
- CNS:
-
Central nervous system
- EAE:
-
Experimental autoimmune encephalomyelitis
- GAPDH:
-
Glyceraldehyde 3-phosphate dehydrogenase
- GFAP:
-
Glial fibrillary acidic protein
- HRP:
-
Horseradish peroxidase
- Iba-1:
-
Anti-ionized calcium binding adaptor molecule-1
- ICAM-1:
-
Intercellular adhesion molecule-1
- IFN-β:
-
Interferon beta
- IL:
-
Interleukin
- iNOS:
-
Inducible nitric oxide synthase
- KRGE:
-
Korean red ginseng extract
- MBP:
-
Myelin basic protein
- MS:
-
Multiple sclerosis
- MCP-1:
-
Monocyte chemotactic protein-1
- MIP-1α:
-
Macrophage inflammatory protein-1α
- PDGFαR:
-
Platelet-derived growth factor
- PTX:
-
Pertussis toxin
- TCR:
-
T cell receptor
- Th1:
-
T helper 1
- Th2:
-
T helper 2
- TNF-α:
-
Tumor necrosis factor-α
- Tregs:
-
Regulatory T cells
- VCAM-1:
-
Vascular cell adhesion molecule-1
References
Lassmann H, van Horssen J (2011) The molecular basis of neurodegeneration in multiple sclerosis. FEBS Lett 585:3715–3723
McFarland HF, Martin R (2007) Multiple sclerosis: a complicated picture of autoimmunity. Nat Immunol 8:913–919
Ragheb S, Lisak R (1993) Multiple sclerosis: genetic background versus environment. Ann Neurol 34:509–510
Serafini B, Rosicarelli B, Franciotta D, Magliozzi R, Reynolds R, Cinque P, Andreoni L, Trivedi P et al (2007) Dysregulated Epstein-Barr virus infection in the multiple sclerosis brain. J Exp Med 204:2899–2912
Engelhardt B, Ransohoff RM (2012) Capture, crawl, cross: the T cell code to breach the blood–brain barriers. Trends Immunol 33:579–589
Chen SJ, Wang YL, Fan HC, Lo WT, Wang CC, Sytwu HK (2012) Current status of the immunomodulation and immunomediated therapeutic strategies for multiple sclerosis. Clin Dev Immunol 2012:970789
Cuzzola VF, Palella E, Celi D, Barresi M, Giacoppo S, Bramanti P, Marino S (2012) Pharmacogenomic update on multiple sclerosis: a focus on actual and new therapeutic strategies. Pharmacogenomics J 12:453–461
Neilley LK, Goodin DS, Goodkin DE, Hauser SL (1996) Side effect profile of interferon beta-1b in MS: results of an open label trial. Neurology 46:552–554
Gasperini C, Ruggieri S (2009) New oral drugs for multiple sclerosis. Neurol Sci 30(Suppl 2):S179–S183
Minagar A (2013) Current and future therapies for multiple sclerosis. Scientifica 2013:249101
Cho I (2012) Effects of Panax ginseng in neurodegenerative diseases. J Ginseng Res 36:342–353
Shibata S, Fujita M, Itokawa H, Tanaka O, Ishii T (1963) Studies on the constituents of Japanese and Chinese crude drugs. XI. Panaxadiol, a sapogenin of ginseng roots. Chem Pharm Bull (Tokyo) 11:759–761
Kaneko H, Nakanishi K (2004) Proof of the mysterious efficacy of ginseng: basic and clinical trials: clinical effects of medical ginseng, Korean red ginseng: specifically, its anti-stress action for prevention of disease. J Pharmacol Sci 95:158–162
Karmazyn M, Moey M, Gan XT (2011) Therapeutic potential of ginseng in the management of cardiovascular disorders. Drugs
Kim HJ, Kim P, Shin CY (2013) A comprehensive review of the therapeutic and pharmacological effects of ginseng and ginsenosides in central nervous system. J Ginseng Res 37(1):8–29
Lee MS, Yang EJ, Kim JI, Ernst E (2009) Ginseng for cognitive function in Alzheimer’s disease: a systematic review. J Alzheimers Dis 18:339–344
Van Kampen J, Robertson H, Hagg T, Drobitch R (2003) Neuroprotective actions of the ginseng extract G115 in two rodent models of Parkinson’s disease. Exp Neurol 184:521–529
Jang M, Lee MJ, Kim CS, Cho IH (2013) Korean Red Ginseng Extract attenuates 3-nitropropionic acid-induced Huntington’s-like symptoms. Evid Based Complement Alternat Med 2013:237207
Hwang I, Ahn G, Park E, Ha D, Song JY, Jee Y (2011) An acidic polysaccharide of Panax ginseng ameliorates experimental autoimmune encephalomyelitis and induces regulatory T cells. Immunol Lett 138:169–178
Hwang I, Ha D, Ahn G, Park E, Joo H, Jee Y (2011) Experimental autoimmune encephalomyelitis: association with mutual regulation of RelA (p65)/NF-kappaB and phospho-IkappaB in the CNS. Biochem Biophys Res Commun 411:464–470
Steinman L, Zamvil SS (2006) How to successfully apply animal studies in experimental allergic encephalomyelitis to research on multiple sclerosis. Ann Neurol 60:12–21
Landis SC, Amara SG, Asadullah K, Austin CP, Blumenstein R, Bradley EW, Crystal RG, Darnell RB et al (2012) A call for transparent reporting to optimize the predictive value of preclinical research. Nature 490:187–191
Administration, K.F.a.D. (2007) Korea Food and Drug Administration. Korea Food Code. Mun-young Publishing Co., Seoul
Lee MJ, Jang M, Jung HS, Kim SH, Cho IH (2012) Ethyl pyruvate attenuates formalin-induced inflammatory nociception by inhibiting neuronal ERK phosphorylation. Mol Pain 8:40
Oyagi A, Ogawa K, Kakino M, Hara H (2010) Protective effects of a gastrointestinal agent containing Korean red ginseng on gastric ulcer models in mice. BMC Complement Altern Med 10:45
Lee MJ, Jang M, Choi J, Lee G, Min HJ, Chung WS, Kim JI, Jee Y, Chae Y, Kim SH, Lee SJ, Cho IH (2015) Bee venom acupuncture alleviates experimental autoimmune encephalomyelitis by upregulating regulatory T cells and suppressing Th1 and Th17 responses. Mol Neurobiol
Jang M, Lee MJ, Cho IH (2014) Ethyl pyruvate ameliorates 3-nitropropionic acid-induced striatal toxicity through anti-neuronal cell death and anti-inflammatory mechanisms. Brain Behav Immun 38:151–165
Fissolo N, Costa C, Nurtdinov RN, Bustamante MF, Llombart V, Mansilla MJ, Espejo C, Montalban X et al (2012) Treatment with MOG-DNA vaccines induces CD4 + CD25 + FoxP3+ regulatory T cells and up-regulates genes with neuroprotective functions in experimental autoimmune encephalomyelitis. J Neuroinflammation 9:139
Piao ZG, Cho IH, Park CK, Hong JP, Choi SY, Lee SJ, Lee S, Park K et al (2006) Activation of glia and microglial p38 MAPK in medullary dorsal horn contributes to tactile hypersensitivity following trigeminal sensory nerve injury. Pain 121:219–231
VanGuilder HD, Bixler GV, Brucklacher RM, Farley JA, Yan H, Warrington JP, Sonntag WE, Freeman WM (2011) Concurrent hippocampal induction of MHC II pathway components and glial activation with advanced aging is not correlated with cognitive impairment. J Neuroinflammation 8:138
Cho IH, Hong J, Suh EC, Kim JH, Lee H, Lee JE, Lee S, Kim CH et al (2008) Role of microglial IKKbeta in kainic acid-induced hippocampal neuronal cell death. Brain 131:3019–3033
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408
Hong J, Cho IH, Kwak KI, Suh EC, Seo J, Min HJ, Choi SY, Kim CH et al (2011) Microglial Toll-like receptor 2 contributes to kainic acid-induced glial activation and hippocampal neuronal cell death. J Biol Chem 285:39447–39457
Fujita H, Nograles KE, Kikuchi T, Gonzalez J, Carucci JA, Krueger JG (2009) Human Langerhans cells induce distinct IL-22-producing CD4+ T cells lacking IL-17 production. Proc Natl Acad Sci U S A 106:21795–21800
Sedgwick JD, Schwender S, Imrich H, Dorries R, Butcher GW, ter Meulen V (1991) Isolation and direct characterization of resident microglial cells from the normal and inflamed central nervous system. Proc Natl Acad Sci U S A 88:7438–7442
Zhu D, Liu M, Yang Y, Ma L, Jiang Y, Zhou L, Huang Q, Pi R et al (2014) Ginsenoside Rd ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice. J Neurosci Res 92:1217–1226
Lobsiger CS, Cleveland DW (2007) Glial cells as intrinsic components of non-cell-autonomous neurodegenerative disease. Nat Neurosci 10:1355–1360
Kim JH, Kim S, Yoon IS, Lee JH, Jang BJ, Jeong SM, Lee JH, Lee BH et al (2005) Protective effects of ginseng saponins on 3-nitropropionic acid-induced striatal degeneration in rats. Neuropharmacology 48:743–756
Willis CL (2010) Glia-induced reversible disruption of blood–brain barrier integrity and neuropathological response of the neurovascular unit. Toxicol Pathol 39:172–185
Hohlfeld R (1997) Biotechnological agents for the immunotherapy of multiple sclerosis. Principles, problems and perspectives. Brain 120(Pt 5):865–916
El-behi M, Rostami A, Ciric B (2010) Current views on the roles of Th1 and Th17 cells in experimental autoimmune encephalomyelitis. J Neuroimmune Pharmacol 5:189–197
Read S, Malmstrom V, Powrie F (2000) Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation. J Exp Med 192:295–302
Kennedy KJ, Strieter RM, Kunkel SL, Lukacs NW, Karpus WJ (1998) Acute and relapsing experimental autoimmune encephalomyelitis are regulated by differential expression of the CC chemokines macrophage inflammatory protein-1alpha and monocyte chemotactic protein-1. J Neuroimmunol 92:98–108
Park JS, Park EM, Kim DH, Jung K, Jung JS, Lee EJ, Hyun JW, Kang JL et al (2009) Anti-inflammatory mechanism of ginseng saponins in activated microglia. J Neuroimmunol 209:40–49
Shin T, Ahn M, Matsumoto Y (2012) Mechanism of experimental autoimmune encephalomyelitis in Lewis rats: recent insights from macrophages. Anatomy Cell Biol 45:141–148
Bowie LE, Roscoe WA, Lui EM, Smith R, Karlik SJ (2012) Effects of an aqueous extract of North American ginseng on MOG(35–55)-induced EAE in mice. Can J Physiol Pharmacol 90:933–939
Attele AS, Wu JA, Yuan CS (1999) Ginseng pharmacology: multiple constituents and multiple actions. Biochem Pharmacol 58:1685–1693
Yuan CS, Wang CZ, Wicks SM, Qi LW (2010) Chemical and pharmacological studies of saponins with a focus on American ginseng. J Ginseng Res 34:160–167
Kim DH (2012) Chemical diversity of Panax ginseng, Panax quinquifolium, and Panax notoginseng. J Ginseng Res 36:1–15
Kemper KJ (2007) The yin and yang of integrative clinical care, education, and research. Explore (New York, NY) 3:37–41
Dan B, Steven C, Erich S, Andrew G (2004) Chinese herbal medicine. Mater Med
Chen X, Ma X, Jiang Y, Pi R, Liu Y, Ma L (2011) The prospects of minocycline in multiple sclerosis. J Neuroimmunol 235:1–8
Guo X, Nakamura K, Kohyama K, Harada C, Behanna HA, Watterson DM, Matsumoto Y, Harada T (2007) Inhibition of glial cell activation ameliorates the severity of experimental autoimmune encephalomyelitis. Neurosci Res 59:457–466
Starossom SC, Mascanfroni ID, Imitola J, Cao L, Raddassi K, Hernandez SF, Bassil R, Croci DO et al (2012) Galectin-1 deactivates classically activated microglia and protects from inflammation-induced neurodegeneration. Immunity 37:249–263
Fabis MJ, Scott GS, Kean RB, Koprowski H, Hooper DC (2007) Loss of blood–brain barrier integrity in the spinal cord is common to experimental allergic encephalomyelitis in knockout mouse models. Proc Natl Acad Sci U S A 104:5656–5661
Zeng Y, Gu B, Ji X, Ding X, Song C, Wu F (2007) Sinomenine, an antirheumatic alkaloid, ameliorates clinical signs of disease in the Lewis rat model of acute experimental autoimmune encephalomyelitis. Biol Pharm Bull 30:1438–1444
Ponomarev ED, Veremeyko T, Barteneva N, Krichevsky AM, Weiner HL (2011) MicroRNA-124 promotes microglia quiescence and suppresses EAE by deactivating macrophages via the C/EBP-alpha-PU.1 pathway. Nat Med 17:64–70
Ransohoff RM, Kivisakk P, Kidd G (2003) Three or more routes for leukocyte migration into the central nervous system. Nat Rev Immunol 3:569–581
Steiner O, Coisne C, Cecchelli R, Boscacci R, Deutsch U, Engelhardt B, Lyck R (2010) Differential roles for endothelial ICAM-1, ICAM-2, and VCAM-1 in shear-resistant T cell arrest, polarization, and directed crawling on blood–brain barrier endothelium. J Immunol 185:4846–4855
Chaudhary P, Marracci GH, Bourdette DN (2006) Lipoic acid inhibits expression of ICAM-1 and VCAM-1 by CNS endothelial cells and T cell migration into the spinal cord in experimental autoimmune encephalomyelitis. J Neuroimmunol 175:87–96
Kan QC, Zhu L, Liu N, Zhang GX (2013) Matrine suppresses expression of adhesion molecules and chemokines as a mechanism underlying its therapeutic effect in CNS autoimmunity. Immunol Res 56:189–196
Ortiz GG, Pacheco-Moises FP, Macias-Islas MA, Flores-Alvarado LJ, Mireles-Ramirez MA, Gonzalez-Renovato ED, Hernandez-Navarro VE, Sanchez-Lopez AL et al (2014) Role of the blood–brain barrier in multiple sclerosis. Arch Med Res 45:687–697
Flynn KM, Michaud M, Madri JA (2013) CD44 deficiency contributes to enhanced experimental autoimmune encephalomyelitis: a role in immune cells and vascular cells of the blood–brain barrier. Am J Pathol 182:1322–1336
Kim H, Ahn M, Choi S, Kim M, Sim KB, Kim J, Moon C, Shin T (2013) Potential role of fibronectin in microglia/macrophage activation following cryoinjury in the rat brain: an immunohistochemical study. Brain Res 1502:11–19
Muzio L, Cavasinni F, Marinaro C, Bergamaschi A, Bergami A, Porcheri C, Cerri F, Dina G et al (2010) Cxcl10 enhances blood cells migration in the sub-ventricular zone of mice affected by experimental autoimmune encephalomyelitis. Mol Cell Neurosci 43:268–280
Wolburg-Buchholz K, Mack AF, Steiner E, Pfeiffer F, Engelhardt B, Wolburg H (2009) Loss of astrocyte polarity marks blood–brain barrier impairment during experimental autoimmune encephalomyelitis. Acta Neuropathol 118:219–233
Compston A, Coles A (2008) Multiple sclerosis. Lancet 372:1502–1517
Reddy J, Illes Z, Zhang X, Encinas J, Pyrdol J, Nicholson L, Sobel RA, Wucherpfennig KW et al (2004) Myelin proteolipid protein-specific CD4 + CD25+ regulatory cells mediate genetic resistance to experimental autoimmune encephalomyelitis. Proc Natl Acad Sci U S A 101:15434–15439
Fontenot JD, Gavin MA, Rudensky AY (2003) Foxp3 programs the development and function of CD4 + CD25+ regulatory T cells. Nat Immunol 4:330–336
Chen X, Oppenheim JJ, Winkler-Pickett RT, Ortaldo JR, Howard OM (2006) Glucocorticoid amplifies IL-2-dependent expansion of functional FoxP3(+)CD4(+)CD25(+) T regulatory cells in vivo and enhances their capacity to suppress EAE. Eur J Immunol 36:2139–2149
Liu YM, Liu XJ, Bai SS, Mu LL, Kong QF, Sun B, Wang DD, Wang JH et al (2010) The effect of electroacupuncture on T cell responses in rats with experimental autoimmune encephalitis. J Neuroimmunol 220:25–33
Le Y, Zhou Y, Iribarren P, Wang J (2004) Chemokines and chemokine receptors: their manifold roles in homeostasis and disease. Cell Mol Immunol 1:95–104
Szczucinski A, Losy J (2007) Chemokines and chemokine receptors in multiple sclerosis. Potential targets for new therapies. Acta Neurol Scand 115:137–146
Giraud SN, Caron CM, Pham-Dinh D, Kitabgi P, Nicot AB (2010) Estradiol inhibits ongoing autoimmune neuroinflammation and NFkappaB-dependent CCL2 expression in reactive astrocytes. Proc Natl Acad Sci U S A 107:8416–8421
Singh NP, Hegde VL, Hofseth LJ, Nagarkatti M, Nagarkatti P (2007) Resveratrol (trans-3,5,4′-trihydroxystilbene) ameliorates experimental allergic encephalomyelitis, primarily via induction of apoptosis in T cells involving activation of aryl hydrocarbon receptor and estrogen receptor. Mol Pharmacol 72:1508–1521
Zhang F, Wei W, Chai H, Xie X (2013) Aurintricarboxylic acid ameliorates experimental autoimmune encephalomyelitis by blocking chemokine-mediated pathogenic cell migration and infiltration. J Immunol 190:1017–1025
Acknowledgments
This research was supported by grants from the Korean Society of Ginseng and the Korea Ginseng Cooperation (2012–2013) and by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and future Planning (2007–0054931 and 2014R1A2A1A11051240). This research was also supported by the Traditional Korean Medicine R&D program founded by the Ministry of Health & Welfare through the Korea Health Industry Development Institute (KHIDI) (HI13C0263).
Conflict of Interest
All the authors of this manuscript have no conflict of interest in this subject.
Authors’ Contributions
MJL performed the behavioral experiment, immunohistochemistry, PCR analysis, flow cytometry, and Western blots and prepared the figures. MJ and JHC assisted with behavioral and histological experiments and prepared the figures. BSC performed the histological experiment associated with semithin section. DYK, SKO, SHK, YSK, SO, JHL, BJC, and SYN commented about the treatment with KRGE, contributed to the interpretation of data, and supervised the project. IHC conceived all experiments, analyzed the results, and wrote the manuscript. All authors have read and approved the final manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, M.J., Jang, M., Choi, J. et al. Korean Red Ginseng and Ginsenoside-Rb1/-Rg1 Alleviate Experimental Autoimmune Encephalomyelitis by Suppressing Th1 and Th17 Cells and Upregulating Regulatory T Cells. Mol Neurobiol 53, 1977–2002 (2016). https://doi.org/10.1007/s12035-015-9131-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12035-015-9131-4