Oral Administration of Gintonin Attenuates Cholinergic Impairments by Scopolamine, Amyloid-β Protein, and Mouse Model of Alzheimer's Disease

  • Kim, Hyeon-Joong (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Shin, Eun-Joo (Neuropsychopharmacology and Toxicology Program, College of Pharmacy, Kangwon National University) ;
  • Lee, Byung-Hwan (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Choi, Sun-Hye (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Jung, Seok-Won (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Cho, Ik-Hyun (Department of Convergence Medical Science, Brain Korea 21 Plus Program, and Institute of Oriental Medicine) ;
  • Hwang, Sung-Hee (Department of Pharmaceutical Engineering, Sangji University) ;
  • Kim, Joon Yong (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Han, Jung-Soo (Department of Biological Sciences, Konkuk University) ;
  • Chung, ChiHye (Department of Biological Sciences, Konkuk University) ;
  • Jang, Choon-Gon (Department of Pharmacology, College of Pharmacy, Sungkyunkwan University) ;
  • Rhim, Hyewon (Center for Neuroscience, Korea Institute of Science and Technology Seoul) ;
  • Kim, Hyoung-Chun (Neuropsychopharmacology and Toxicology Program, College of Pharmacy, Kangwon National University) ;
  • Nah, Seung-Yeol (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University)
  • Received : 2015.05.04
  • Accepted : 2015.06.29
  • Published : 2015.09.30


Gintonin is a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand. Oral administration of gintonin ameliorates learning and memory dysfunctions in Alzheimer's disease (AD) animal models. The brain cholinergic system plays a key role in cognitive functions. The brains of AD patients show a reduction in acetylcholine concentration caused by cholinergic system impairments. However, little is known about the role of LPA in the cholinergic system. In this study, we used gintonin to investigate the effect of LPA receptor activation on the cholinergic system in vitro and in vivo using wild-type and AD animal models. Gintonin induced $[Ca^{2+}]_i $ transient in cultured mouse hippocampal neural progenitor cells (NPCs). Gintonin-mediated $[Ca^{2+}]_i $ transients were linked to stimulation of acetylcholine release through LPA receptor activation. Oral administration of gintonin-enriched fraction (25, 50, or 100 mg/kg, 3 weeks) significantly attenuated scopolamine-induced memory impairment. Oral administration of gintonin (25 or 50 mg/kg, 1 2 weeks) also significantly attenuated amyloid-${\beta}$ protein ($A{\beta}$)-induced cholinergic dysfunctions, such as decreased acetylcholine concentration, decreased choline acetyltransferase (ChAT) activity and immunoreactivity, and increased acetylcholine esterase (AChE) activity. In a transgenic AD mouse model, long-term oral administration of gintonin (25 or 50 mg/kg, 3 months) also attenuated AD-related cholinergic impairments. In this study, we showed that activation of G protein-coupled LPA receptors by gintonin is coupled to the regulation of cholinergic functions. Furthermore, this study showed that gintonin could be a novel agent for the restoration of cholinergic system damages due to $A{\beta}$ and could be utilized for AD prevention or therapy.


Supported by : Ministry of Education, Science, and Technology


  1. Alzheimer, A. (1907). Uber eine eigenartige Erkrankung der Hirnrinde. Allg. Zeitschrife Psychiatr. 64, 146-148.
  2. Bales, K.R., Tzavara, E.T., Wu, S., Wade, M.R., Bymaster, F.P., Paul, S.M., and Nomikos, G.G. (2006). Cholinergic dysfunction in a mouse model of Alzheimer disease is reversed by an anti-A beta antibody. J. Clin. Invest. 116, 825-832.
  3. Bartus, R.T., Dean, R.L., Beer, B., and Lippa, A.S. (1982). The cholinergic hypothesis of geriatric memory dysfunction. Science 217, 408-414.
  4. Braak, H., Braak, E., Grundke-Iqbal, I., and Iqbal, K. (1986). Occurrence of neuropil threads in the senile human brain and in Alzheimer's disease: a third location of paired helical filaments outside of neurofibrillary tangles and neuritic plaques. Neurosci. Lett. 65, 351-355.
  5. Castilla-Ortega, E., Sanchez-Lopez, J., Hoyo-Becerra, C., Matas-Rico, E., Zambrana-Infantes, E., Chun, J., De Fonseca, F.R., Pedraza, C., Estivill-Torrus, G., and Santin, L.J. (2010). Exploratory, anxiety and spatial memory impairments are dissociated in mice lacking the LPA1 receptor. Neurobiol. Learn. Mem. 94, 73-82.
  6. Castilla-Ortega, E., Hoyo-Becerra, C., Pedraza, C., Chun, J., Rodriguez De Fonseca, F., Estivill-Torrus, G., and Santin, L.J. (2011). Aggravation of chronic stress effects on hippocampal neurogenesis and spatial memory in LPA1 receptor knockout mice. PLoS One 6, e25522.
  7. Castilla-Ortega, E., Pedraza, C., Chun, J., de Fonseca, F.R., Estivill-Torrus, G., and Santin, L.J. (2012). Hippocampal c-Fos activation in normal and LPA-null mice after two object recognition tasks with different memory demands. Behav. Brain Res. 232, 400-405.
  8. Chao, L.P., and Wolfgram, F. (1972). Spectrophotometric assay for choline acetyltransferase. Anal. Biochem. 46, 114-118.
  9. Choi, S.H., Jung, S.W., Kim, H.S., Kim, H.J., Lee B.H., Kim, J.Y., Kim J.H., Hwang, S.H., Rhim, H., Kim, HC., et al. (2015a). A brief method for the preparation of gintonin-enriched fraction from ginseng. J. Ginseng Res. doi:10.1016/j.jgr.2015.05.002 (in press).
  10. Choi, S.H., Hong, M.K., Kim, H.J., Ryoo, N., Rhim, H., Nah, S.Y., and Kang, L.W. (2015b). Crystal structure of ginseng major latex-like protein 151 and its proposed lysophosphatidic acidbinding mechanism. Acta Crystallogr. D Biol. Crystallogr. 71, 1039-1050.
  11. Colom, L.V., Castaneda, M.T., Hernandez, S., Perry, G., Jaime, S., Touhami, A. (2011). Intrahippocampal amyloid-${\beta}$ (1-40) injections injure medial septal neurons in rats. Curr. Alzheimer Res. 8, 832-840.
  12. Cui, H.L., and Qiao, J.T. (2006). Promotive action of lysophosphatidic acid on proliferation of rat embryonic neural stem cells and their differentiation to cholinergic neurons in vitro. Sheng Li Xue Bao 58, 547-555.
  13. Dash, P.K., Orsi, S.A., Moody, M., and Moore, A.N. (2004). A role for hippocampal Rho-ROCK pathway in long-term spatial memory. Biochem. Biophys. Res. Commun. 322, 893-898.
  14. Davie, B.J., Christopoulos, A., and Scammells, P.J. (2013). Development of M1 mAChR allosteric and bitopic ligands: prospective therapeutics for the treatment of cognitive deficits. ACS Chem. Neurosci. 4, 1026-1048.
  15. Dubin, A.E., Bahnson, T., Weiner, J.A., Fukushima, N., and Chun, J. (1999). Lysophosphatidic acid stimulates neurotransmitter-like conductance changes that precede GABA and L-glutamate in early, presumptive cortical neuroblasts. J. Neurosci. 19, 1371-1381.
  16. Van Der Flier, W.M., Van Den Heuvel, D.M.J., Weverling-Rijnsburger, A.W.E., Spilt, A., Bollen, E.L.E.M., Westendorp, R.G.J., Middelkoop, H.A.M., and Van Buchem, M.A. (2002). Cognitive decline in AD and mild cognitive impairment is associated with global brain damage. Neurology 59, 874-879.
  17. Hasebe, N., Fujita, Y., Ueno, M., Yoshimura, K., Fujino, Y., and Yamashita, T. (2013). Soluble ${\beta}$-amyloid Precursor Protein Alpha binds to p75 neurotrophin receptor to promote neurite outgrowth. PLoS One 8, e82321.
  18. Hecht, J.H., Weiner, J.A., Post, S.R., and Chun, J. (1996). Ventricular zone gene-1 (vzg-1) encodes a lysophosphatidic acid receptor expressed in neurogenic regions of the developing cerebral cortex. J. Cell Biol. 135, 1071-1083.
  19. Heo, J.H., Lee, S.T., Chu, K., Oh, M.J., Park, H.J., Shim, J.Y., and Kim, M. (2008). An open-label trial of Korean red ginseng as an adjuvant treatment for cognitive impairment in patients with Alzheimer's disease. Eur. J. Neurol. 15, 865-868.
  20. Hooijmans, C.R., Rutters, F., Dederen, P.J., Gambarota, G., Veltien, A., van Groen, T., Broersen, L.M., Lutjohann, D., Heerschap, A., Tanila, H., et al. (2007). Changes in cerebral blood volume and amyloid pathology in aged Alzheimer APP/PS1 mice on a docosahexaenoic acid (DHA) diet or cholesterol enriched Typical Western Diet (TWD). Neurobiol. Dis. 28, 16-29.
  21. Hwang, S.H., Shin, T.J., Choi, S.H., Cho, H.J., Lee, B.H., Pyo, M.K., Lee, J.H., Kang, J., Kim, H.J., Park, C.W., et al. (2012a). Gintonin, newly identified compounds from ginseng, is novel lysophosphatidic acids-protein complexes and activates G protein-coupled lysophosphatidic acid receptors with high affinity. Mol. Cells 33, 151-162.
  22. Hwang, S.H., Shin, E.J., Shin, T.J., Lee, B.H., Choi, S.H., Kang, J., Kim, H.J., Kwon, S.H., Jang, C.G., Lee, J.H., et al. (2012b). Gintonin, a ginseng-derived lysophosphatidic acid receptor ligand, attenuates alzheimer's disease-related neuropathies: Involvement of non-amyloidogenic processing. J. Alzheimer's Dis. 31, 207-223.
  23. Hwang, S.H., Lee, B.H., Choi, S.H., Kim, H.J., Jung, S.W., Kim, H.S., Shin, H.C., Park, H.J., Park, K.H., Lee, M.K., et al. (2015). Gintonin, a novel ginseng-derived lysophosphatidic acid receptor ligand, stimulates neurotransmitter release. Neurosci. Lett. 584, 356-361.
  24. Jaffar, S., Counts, S.E., Ma, S.Y., Dadko, E., Gordon, M.N., Morgan, D., and Mufson, E.J. (2001) Neuropathology of mice carrying mutant APP(swe) and/or PS1(M146L) transgenes: alterations in the p75(NTR) cholinergic basal forebrain septohippocampal pathway. Exp. Neurol. 170, 227-243.
  25. Jin, C.H., Shin, E.J., Park, J.B., Jang, C.G., Li, Z., Kim, M.S., Koo, K.H., Yoon, H.J., Park, S.-J., Choi, W.-C., et al. (2009). Fustin flavonoid attenuates beta-amyloid (1-42)-induced learning impairment. J. Neurosci. Res. 87, 3658-3670.
  26. Jung, B.D., Shin, E.J., Nguyen, X.K.T., Jin, C.H., Bach, J.H., Park, S.J., Nah, S.Y., Wie, M.B., Bing, G., and Kim, H.C. (2010). Potentiation of methamphetamine neurotoxicity by intrastriatal lipopolysaccharide administration. Neurochem. Int. 56, 229-244.
  27. Kennedy, D.O., and Scholey, A.B. (2003). Ginseng: potential for the enhancement of cognitive performance and mood. Pharmacol. Biochem. Behav. 75, 687-700.
  28. Kim, J.H., Choi, S., Jung, J.E., Roh, E.J., and Kim, H.J. (2006). Capacitative Ca2+ entry is involved in regulating soluble amyloid precursor protein (sAPPalpha) release mediated by muscarinic acetylcholine receptor activation in neuroblastoma SH-SY5Y cells. J. Neurochem. 97, 245-254.
  29. Kim, B.-W., Yang, S., Lee, C.H., and Son, H. (2011). A critical time window for the survival of neural progenitor cells by HDAC inhibitors in the hippocampus. Mol. Cells 31, 159-164.
  30. Kim, E.J., Jung, I.H., Van Le, T.K., Jeong, J.J., Kim, N.J., and Kim, D.H. (2013). Ginsenosides Rg5 and Rh3 protect scopolamineinduced memory deficits in mice. J. Ethnopharmacol. 146, 294-299.
  31. Lee, S.T., Chu, K., Sim, J.Y., Heo, J.H., and Kim, M. (2008). Panax ginseng enhances cognitive performance in Alzheimer disease. Alzheimer Dis. Assoc. Disord. 22, 222-226.
  32. Liu, J.X., Cong, W.H., Xu, L., and Wang, J.N. (2004). Effect of combination of extracts of ginseng and ginkgo biloba on acetylcholine in amyloid beta-protein-treated rats determined by an improved HPLC. Acta Pharmacol. Sin. 25, 1118-1123.
  33. Maurice, T., Lockhart, B.P., and Privat, A. (1996). Amnesia induced in mice by centrally administered beta-amyloid peptides involves cholinergic dysfunction. Brain Res. 706, 181-193.
  34. Pakaski, M., and Kalman, J. (2008). Interactions between the amyloid and cholinergic mechanisms in Alzheimer's disease. Neurochem. Int. 53, 103-111.
  35. Park, H., Kim, S., Rhee, J., Kim, H.J., Han, J.S., Nah, S.Y., and Chung, C. (2015). Synaptic enhancement induced by gintonin via lysophosphatidic acid receptor activation in central synapses. J. Neurophysiol. 113, 1493-1500.
  36. Pedraza, C., Sanchez-Lopez, J., Castilla-Ortega, E., Rosell-Valle, C., Zambrana-Infantes, E., Garcia-Fernandez, M., Rodriguez de Fonseca, F., Chun, J., Santin, L.J., and Estivill-Torrus, G. (2014). Fear extinction and acute stress reactivity reveal a role of LPA(1) receptor in regulating emotional-like behaviors. Brain Struct. Funct. 219, 1659-1672.
  37. Perez, S.E., Dar, S., Ikonomovic, M.D., DeKosky, S.T., and Mufson, E.J. (2007). Cholinergic Forebrain Degeneration in the APPswe/$PS1{\Delta}E9$ Transgenic Mouse. Neurobiol. Dis. 28, 3-15.
  38. Pyo, M.K., Choi, S.-H., Hwang, S.H., Shin, T.J., Lee, B.H., Lee, S.M., Lim, Y.H., Kim, D.H., and Nah, S.Y. (2011). Novel Glycolipoproteins from Ginseng. J. Ginseng Res. 35, 92-103.
  39. Shin, T.J., Kim, H.J., Kwon, B.J., Choi, S.H., Kim, H.B., Hwang, S.H., Lee, B.H., Lee, S.M., Zukin, R.S., Park, J.H., et al. (2012). Gintonin, a ginseng-derived novel ingredient, evokes long-term potentiation through N-methyl-D-aspartic acid receptor activation: involvement of LPA receptors. Mol. Cells 34, 563-572.
  40. Shiono, S., Kawamoto, K., Yoshida, N., Kondo, T., and Inagami, T. (1993). Neurotransmitter release from lysophosphatidic acid stimulated PC12 cells: involvement of lysophosphatidic acid receptors. Biochem. Biophys. Res. Commun. 193, 667-673.
  41. Su, C.F., Cheng, J.T., and Liu, I.M. (2007). Increase of acetylcholine release by Panax ginseng root enhances insulin secretion in Wistar rats. Neurosci. Lett. 412, 101-104.
  42. Sun, Y., Nam, J.S., Han, D.H., Kim, N.H., Choi, H.K., Lee, J.K., Rhee, H.J., and Huh, S.O. (2010). Lysophosphatidic acid induces upregulation of Mcl-1 and protects apoptosis in a PTXdependent manner in H19-7 cells. Cell. Signal. 22, 484-494.
  43. Tabuchi, S., Kume, K., Aihara, M., and Shimizu, T. (2000). Expression of lysophosphatidic acid receptor in rat astrocytes: mitogenic effect and expression of neurotrophic genes. Neurochem Res. 25, 573-582.
  44. Trimbuch, T., Beed, P., Vogt, J., Schuchmann, S., Maier, N., Kintscher, M., Breustedt, J., Schuelke, M., Streu, N., Kieselmann, O., et al. (2009). Synaptic PRG-1 modulates excitatory transmission via lipid phosphate-mediated signaling. Cell 138, 1222-1235.
  45. Vassar, R., Bennett, B.D., Babu-Khan, S., Kahn, S., Mendiaz, E.A., Denis, P., Teplow, D.B., Ross, S., Amarante, P., Loeloff, R., et al. (1999). Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science 286, 735-741.
  46. Wang, Q., Shin, E.J., Nguyen, X.K., Li, Q., Bach, J.H., Bing, G., Kim, W.K., and Kim, H.C., Hong, J.S. (2012). Endogenous dynorphin protects against neurotoxin-elicited nigrostriatal dopaminergic neuron damage and motor deficits in mice. J. Neuroinflammation. 9, 124.
  47. Wong, T.P., Debeir, T., Duff, K., and Cuello, A.C. (1999). Reorganization of cholinergic terminals in the cerebral cortex and hippocampus in transgenic mice carrying mutated presenilin-1 and amyloid precursor protein transgenes. J. Neurosci. 19, 2706-2716.
  48. Yankner, B.A. (1996). Mechanisms of neuronal degeneration in Alzheimer's disease. Neuron 16, 921-932.

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