Journal of Ginseng Research

  • 제44권1호
  • /
  • Pages.67-78
  • /
  • 2020
  • /
  • 1226-8453(pISSN)
  • /
  • 2093-4947(eISSN)
고려인삼학회 (The Korean Society of Ginseng)
권호 표지
DOI QR코드

DOI QR Code

Gintonin stimulates autophagic flux in primary cortical astrocytes

  • Rahman, Md. Ataur (Center for Neuroscience, Korea Institute of Science and Technology) ;
  • Hwang, Hongik (Center for Neuroscience, Korea Institute of Science and Technology) ;
  • Nah, Seung-Yeol (Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University) ;
  • Rhim, Hyewhon (Center for Neuroscience, Korea Institute of Science and Technology)
  • 투고 : 2018.03.29
  • 심사 : 2018.08.08
  • 발행 : 2020.01.15
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Background: Gintonin (GT), a novel ginseng-derived exogenous ligand of lysophosphatidic acid (LPA) receptors, has been shown to induce cell proliferation and migration in the hippocampus, regulate calcium-dependent ion channels in the astrocytes, and reduce β-amyloid plaque in the brain. However, whether GT influences autophagy in cortical astrocytes is not yet investigated. Methods: We examined the effect of GT on autophagy in primary cortical astrocytes using immunoblot and immunocytochemistry assays. Suppression of specific proteins was performed via siRNA. LC3 puncta was determined using confocal microscopy. Results: GT strongly upregulated autophagy marker LC3 by a concentration- as well as time-dependent manner via G protein-coupled LPA receptors. GT-induced autophagy was further confirmed by the formation of LC3 puncta. Interestingly, on pretreatment with an mammalian target of rapamycin (mTOR) inhibitor, rapamycin, GT further enhanced LC3-II and LC3 puncta expression. However, GT-induced autophagy was significantly attenuated by inhibition of autophagy by 3-methyladenine and knockdown Beclin-1, Atg5, and Atg7 gene expression. Importantly, when pretreated with a lysosomotropic agent, E-64d/peps A or bafilomycin A1, GT significantly increased the levels of LC3-II along with the formation of LC3 puncta. In addition, GT treatment enhanced autophagic flux, which led to an increase in lysosome-associated membrane protein 1 and degradation of ubiquitinated p62/SQSTM1. Conclusion: GT induces autophagy via mTOR-mediated pathway and elevates autophagic flux. This study demonstrates that GT can be used as an autophagy-inducing agent in cortical astrocytes.

키워드

참고문헌

  1. Mizushima N, Levine B, Cuervo AM, Klionsky DJ. Autophagy fights disease through cellular self-digestion. Nature 2008;451:1069-75. https://doi.org/10.1038/nature06639
  2. Komatsu M, Waguri S, Chiba T, Murata S, Iwata J, Tanida I, Ueno T, Koike M, Uchiyama Y, Kominami E, et al. Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature 2006;441:880-4. https://doi.org/10.1038/nature04723
  3. Singh R, Kaushik S, Wang Y, Xiang Y, Novak I, Komatsu M, Tanaka K, Cuervo AM, Czaja MJ. Autophagy regulates lipid metabolism. Nature 2009;458:1131-5. https://doi.org/10.1038/nature07976
  4. Rubinsztein DC, Marino G, Kroemer G. Autophagy and aging. Cell 2011;146:682-95. https://doi.org/10.1016/j.cell.2011.07.030
  5. Toth ML, Sigmond T, Borsos E, Barna J, Erdelyi P, Takacs-Vellai K, Orosz L, Kovacs AL, Csikos G, Sass M, et al. Longevity pathways converge on autophagy genes to regulate life span in Caenorhabditis elegans. Autophagy 2008;4:330-8. https://doi.org/10.4161/auto.5618
  6. Simonsen A, Cumming RC, Brech A, Isakson P, Schubert DR, Finley KD. Promoting basal levels of autophagy in the nervous system enhances longevity and oxidant resistance in adult Drosophila. Autophagy 2008;4:176-84. https://doi.org/10.4161/auto.5269
  7. Meng Q, Cai D. Defective hypothalamic autophagy directs the central pathogenesis of obesity via the IkappaB kinase beta (IKKbeta)/NF-kappaB pathway. J Biol Chem 2011;286:32324-32. https://doi.org/10.1074/jbc.M111.254417
  8. Yang L, Li P, Fu S, Calay ES, Hotamisligil GS. Defective hepatic autophagy in obesity promotes ER stress and causes insulin resistance. Cell Metab 2010;11:467-78. https://doi.org/10.1016/j.cmet.2010.04.005
  9. Jung HS, Lee MS. Role of autophagy in diabetes and mitochondria. Ann. N. Y. Acad. Sci 2010;1201:79-83. https://doi.org/10.1111/j.1749-6632.2010.05614.x
  10. Spilman P, Podlutskaya N, Hart MJ, Debnath J, Gorostiza O, Bredesen D, Richardson A, Strong R, Galvan V. Inhibition of mTOR by rapamycin abolishes cognitive deficits and reduces amyloid-beta levels in a mouse model of Alzheimer's disease. PLoS One 2010;5:e9979. https://doi.org/10.1371/journal.pone.0009979
  11. Rodriguez-Navarro JA, Rodriguez L, Casarejos MJ, Solano RM, Gomez A, Perucho J, Cuervo AM, Garcia de Yebenes J, Mena MA. Trehalose ameliorates dopaminergic and tau pathology in parkin deleted/tau overexpressing mice through autophagy activation. Neurobiol Dis 2010;39:423-38. https://doi.org/10.1016/j.nbd.2010.05.014
  12. Pan T, Kondo S, Le W, Jankovic J. The role of autophagy-lysosome pathway in neurodegeneration associated with Parkinson's disease. Brain 2008;131:1969-78. https://doi.org/10.1093/brain/awm318
  13. Ravikumar B, Vacher C, Berger Z, Davies JE, Luo S, Oroz LG, Scaravilli F, Easton DF, Duden R, O'Kane CJ, et al. Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease. Nat Genet 2004;36:585-95. https://doi.org/10.1038/ng1362
  14. Hara T, Takamura A, Kishi C, Iemura SI, Natsume T, Guan JL, Mizushima N. FIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells. J Cell Biol 2008;181:497-510. https://doi.org/10.1083/jcb.200712064
  15. Hernandez D, Torres CA, Setlik W, Cebrian C, V Mosharov E, Tang G, Cheng HC, Kholodilov N, Yarygina O, Burke RE, et al. Regulation of presynaptic neurotransmission by macroautophagy. Neuron 2012;74:277-84. https://doi.org/10.1016/j.neuron.2012.02.020
  16. Komatsu M, Wang QJ, Holstein GR, Friedrich VLJ, Iwata J, Kominami E, Chait BT, Tanaka K, Yue Z. Essential role for autophagy protein Atg7 in the maintenance of axonal homeostasis and the prevention of axonal degeneration. Proc Natl Acad Sci U S A 2007;104:14489-94. https://doi.org/10.1073/pnas.0701311104
  17. Nixon RA. The role of autophagy in neurodegenerative disease. Nat Med 2013;19:983-97. https://doi.org/10.1038/nm.3232
  18. Choi SH, Jung SW, Lee BH, Kim HJ, Hwang SH, Kim HK, Nah SY. Ginseng pharmacology: a new paradigm based on gintonin-lysophosphatidic acid receptor interactions. Front Pharmacol 2015;6:245.
  19. Shin TJ, Kim HJ, Kwon BJ, Choi SH, Kim HB, Hwang SH, Lee BH, Lee SM, Zukin RS, Park JH, et al. Gintonin, a ginseng-derived novel ingredient, evokes long-term potentiation through N-methyl-D-aspartic acid receptor activation: involvement of LPA receptors. Mol Cells 2012;34:563-72. https://doi.org/10.1007/s10059-012-0254-4
  20. Park H, Kim S, Rhee J, Kim HJ, Han JS, Nah SY, Chung C. Synaptic enhancement induced by gintonin via lysophosphatidic acid receptor activation in central synapses. J Neurophysiol 2015;113:1493-500. https://doi.org/10.1152/jn.00667.2014
  21. Kim S, Kim MS, Park K, Kim HJ, Jung SW, Nah SY, Han JS, Chung C. Hippocampus-dependent cognitive enhancement induced by systemic gintonin administration. J Ginseng Res 2016;40:55-61. https://doi.org/10.1016/j.jgr.2015.05.001
  22. Hwang SH, Shin EJ, Shin TJ, Lee BH, Choi SH, Kang J, Kim HJ, Kwon SH, Jang CG, Lee JH, et al. Gintonin, a ginseng-derived lysophosphatidic acid receptor ligand, attenuates Alzheimer's disease-related neuropathies: involvement of non-amyloidogenic processing. J Alzheimers Dis 2012;31:207-23. https://doi.org/10.3233/JAD-2012-120439
  23. Kim HJ, Shin EJ, Lee BH, Choi SH, Jung SW, Cho IH, Hwang SH, Kim JY, Han JS, Chung C, et al. Oral administration of gintonin attenuates cholinergic impairments by scopolamine, amyloid-beta protein, and mouse model of Alzheimer's disease. Mol Cells 2015;38:796-805. https://doi.org/10.14348/MOLCELLS.2015.0116
  24. Choi SH, Shin TJ, Lee BH, Hwang SH, Kang J, Kim HJ, Park CW, Nah SY. An edible gintonin preparation from ginseng. J Ginseng Res 2011;35:471-8. https://doi.org/10.5142/jgr.2011.35.4.471
  25. Ohta H, Sato K, Murata N, Damirin A, Malchinkhuu E, Kon J, Kimura T, Tobo M, Yamazaki Y, Watanabe T, et al. Ki16425, a subtype-selective antagonist for EDG-family lysophosphatidic acid receptors. Mol Pharmacol 2003;64:994-1005. https://doi.org/10.1124/mol.64.4.994
  26. Eikelenboom P, Hoozemans JJ, Veerhuis R, van Exel E, Rozemuller AJ, van Gool WA. Whether, when and how chronic inflammation increases the risk of developing late-onset Alzheimer's disease. Alzheimers Res Ther 2012;4:15. https://doi.org/10.1186/alzrt118
  27. Law BYK, Wang M, Ma DL, Al-Mousa F, Michelangeli F, Cheng SH, Ng MHL, To KF, Mok AYF, Ko RYY, et al. Alisol B, a novel inhibitor of the sarcoplasmic/endoplasmic reticulum Ca(2+) ATPase pump, induces autophagy, endoplasmic reticulum stress, and apoptosis. Mol Cancer Ther 2010;9:718-30. https://doi.org/10.1158/1535-7163.MCT-09-0700
  28. Mizushima N, Klionsky DJ. Protein turnover via autophagy: implications for metabolism. Annu Rev Nutr 2007;27:19-40. https://doi.org/10.1146/annurev.nutr.27.061406.093749
  29. Shaw RJ. LKB1 and AMP-activated protein kinase control of mTOR signalling and growth. Acta Physiol (Oxf) 2009;196:65-80. https://doi.org/10.1111/j.1748-1716.2009.01972.x
  30. Rahman MA, Bishayee K, Sadra A, Huh SO. Oxyresveratrol activates parallel apoptotic and autophagic cell death pathways in neuroblastoma cells. Biochim Biophys Acta 2017;1861:23-36. https://doi.org/10.1016/j.bbagen.2016.10.025
  31. Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell 2008;132:27-42. https://doi.org/10.1016/j.cell.2007.12.018
  32. Rahman MA, Bishayee K, Habib K, Sadra A, Huh SO. 18alpha-Glycyrrhetinic acid lethality for neuroblastoma cells via de-regulating the Beclin-1/Bcl-2 complex and inducing apoptosis. Biochem Pharmacol 2016;117:97-112. https://doi.org/10.1016/j.bcp.2016.08.006
  33. He C, Levine B. The Beclin 1 interactome. Curr Opin Cell Biol 2010;22:140-9. https://doi.org/10.1016/j.ceb.2010.01.001
  34. Liang XH, Kleeman LK, Jiang HH, Gordon G, Goldman JE, Berry G, Herman B, Levine B. Protection against fatal Sindbis virus encephalitis by beclin, a novel Bcl-2-interacting protein. J Virol 1998;72:8586-96. https://doi.org/10.1128/jvi.72.11.8586-8596.1998
  35. Mizushima N, Yoshimori T, Levine B. Methods in mammalian autophagy research. Cell 2010;140:313-26. https://doi.org/10.1016/j.cell.2010.01.028
  36. Tanida I, Minematsu-Ikeguchi N, Ueno T, Kominami E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 2005;1:84-91. https://doi.org/10.4161/auto.1.2.1697
  37. Mizushima N, Yoshimori T. How to interpret LC3 immunoblotting. Autophagy 2007;3:542-5. https://doi.org/10.4161/auto.4600
  38. Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, Stenmark H, Johansen T. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 2005;171:603-14. https://doi.org/10.1083/jcb.200507002
  39. Bjorkoy G, Lamark T, Pankiv S, Overvatn A, Brech A, Johansen T. Monitoring autophagic degradation of p62/SQSTM1. Methods Enzymol 2009;452:181-97. https://doi.org/10.1016/S0076-6879(08)03612-4
  40. Ma X, Godar RJ, Liu H, Diwan A. Enhancing lysosome biogenesis attenuates BNIP3-induced cardiomyocyte death. Autophagy 2012;8:297-309. https://doi.org/10.4161/auto.18658
  41. Kraft C, Peter M, Hofmann K. Selective autophagy: ubiquitin-mediated recognition and beyond. Nat Cell Biol 2010;12:836-41. https://doi.org/10.1038/ncb0910-836
  42. Im D, Nah S. Yin and Yang of ginseng pharmacology: ginsenosides vs gintonin. Acta Pharmacol Sin 2013;34:1367-73. https://doi.org/10.1038/aps.2013.100
  43. Kim J, Kundu M, Viollet B, Guan KL. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 2011;13:132-41. https://doi.org/10.1038/ncb2152
  44. Sarkar S. Regulation of autophagy by mTOR-dependent and mTORindependent pathways: autophagy dysfunction in neurodegenerative diseases and therapeutic application of autophagy enhancers. Biochem Soc Trans 2013;41:1103-30. https://doi.org/10.1042/BST20130134
  45. Wang Z, Wilson WA, Fujino MA, Roach PJ. Antagonistic controls of autophagy and glycogen accumulation by Snf1p, the yeast homolog of AMP-activated protein kinase, and the cyclin-dependent kinase Pho85p. Mol Cell Biol 2001;21:5742-52. https://doi.org/10.1128/MCB.21.17.5742-5752.2001
  46. Meley D, Bauvy C, Houben-Weerts JHPM, Dubbelhuis PF, Helmond MTJ, Codogno P, Meijer AJ. AMP-activated protein kinase and the regulation of autophagic proteolysis. J Biol Chem 2006;281:34870-9. https://doi.org/10.1074/jbc.M605488200
  47. Neshat MS, Mellinghoff IK, Tran C, Stiles B, Thomas G, Petersen R, Frost P, Gibbons JJ, Wu H, Sawyers CL. Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci U S A 2001;98:10314-9. https://doi.org/10.1073/pnas.171076798
  48. Son SM, Kang S, Choi H, Mook-Jung I. Statins induce insulin-degrading enzyme secretion from astrocytes via an autophagy-based unconventional secretory pathway. Mol Neurodegener 2015;10:56. https://doi.org/10.1186/s13024-015-0054-3
  49. Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, Tokuhisa T, Ohsumi Y, Yoshimori T. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol 2001;152:657-68. https://doi.org/10.1083/jcb.152.4.657
  50. Komatsu M, Waguri S, Ueno T, Iwata J, Murata S, Tanida I, Ezaki J, Mizushima N, Ohsumi Y, Uchiyama Y, et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J Cell Biol 2005;169:425-34. https://doi.org/10.1083/jcb.200412022
  51. Saitoh T, Fujita N, Hayashi T, Takahara K, Satoh T, Lee H, Matsunaga K, Kageyama S, Omori H, Noda T, et al. Atg9a controls dsDNA-driven dynamic translocation of STING and the innate immune response. Proc Natl Acad Sci U S A 2009;106:20842-6. https://doi.org/10.1073/pnas.0911267106
  52. Cadwell K, Liu JY, Brown SL, Miyoshi H, Loh J, Lennerz JK, Kishi C, Kc W, Carrero JA, Hunt S, et al. A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells. Nature 2008;456:259-63. https://doi.org/10.1038/nature07416
  53. Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A, Rosen J, Eskelinen EL, Mizushima N, Ohsumi Y, et al. Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. J Clin Invest 2003;112:1809-20. https://doi.org/10.1172/JCI20039
  54. Fimia GM, Stoykova A, Romagnoli A, Giunta L, Di Bartolomeo S, Nardacci R, Corazzari M, Fuoco C, Ucar A, Schwartz P, et al. Ambra1 regulates autophagy and development of the nervous system. Nature 2007;447:1121-5. https://doi.org/10.1038/nature05925
  55. Huang M, Jiang X, Liang Y, Liu Q, Chen S, Guo Y. Berberine improves cognitive impairment by promoting autophagic clearance and inhibiting production of beta-amyloid in APP/tau/PS1 mouse model of Alzheimer's disease. Exp Gerontol 2017;91:25-33. https://doi.org/10.1016/j.exger.2017.02.004
  56. Mizushima N, Yoshimori T, Ohsumi Y. The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol 2011;27:107-32. https://doi.org/10.1146/annurev-cellbio-092910-154005
  57. Rubinsztein DC, Cuervo AM, Ravikumar B, Sarkar S, Korolchuk V, Kaushik S, Klionsky DJ. In search of an "autophagomometer". Autophagy 2009;5:585-9. https://doi.org/10.4161/auto.5.5.8823
  58. Hwang J, Lee S, Lee JT, Kwon TK, Kim DR, Kim H, Park HC, Suk K. Gangliosides induce autophagic cell death in astrocytes. Br J Pharmacol 2010;159:586-603. https://doi.org/10.1111/j.1476-5381.2009.00563.x
  59. He C, Klionsky DJ. Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 2009;43:67-93. https://doi.org/10.1146/annurev-genet-102808-114910
  60. Fabrizi C, Pompili E, De Vito S, Somma F, Catizone A, Ricci G, Lenzi P, Fornai F, Fumagalli L. Impairment of the autophagic flux in astrocytes intoxicated by trimethyltin. Neurotoxicology 2016;52:12-22. https://doi.org/10.1016/j.neuro.2015.10.004
  61. Eskelinen EL, Tanaka Y, Saftig P. At the acidic edge: emerging functions for lysosomal membrane proteins. Trends Cell Biol 2003;13:137-45. https://doi.org/10.1016/S0962-8924(03)00005-9
  62. Fukuda M. Lysosomal membrane glycoproteins. Structure, biosynthesis, and intracellular trafficking. J Biol Chem 1991;266:21327-30. https://doi.org/10.1016/S0021-9258(18)54636-6
  63. Kim DH. Chemical diversity of Panax ginseng, Panax quinquifolium, and Panax notoginseng. J Ginseng Res 2012;36:1-15. https://doi.org/10.5142/jgr.2012.36.1.1
  64. Pyo MK, Choi SH, Hwang SH, Shin TJ, Lee BH, Lee SM, Lim YH, Kim DH, Nah SY. Novel glycolipoproteins from ginseng. J Ginseng Res 2011;35:92-103. https://doi.org/10.5142/jgr.2011.35.1.092
  65. Lee BH, Choi SH, Kim HJ, Park SD, Rhim H, Kim HC, Hwang SH, Nah SY. Gintonin absorption in intestinal model systems. J Ginseng Res 2018;42:35-41. https://doi.org/10.1016/j.jgr.2016.12.007
  66. Kim DG, Jang M, Choi SH, Kim HJ, Jhun H, Kim HC, Rhim H, Cho IH, Nah SY. Gintonin, a ginseng-derived exogenous lysophosphatidic acid receptor ligand, enhances blood-brain barrier permeability and brain delivery. Int J Biol Macromol 2018;114:1325-37. https://doi.org/10.1016/j.ijbiomac.2018.03.158

피인용 문헌

  1. Molecular Insights Into Therapeutic Potential of Autophagy Modulation by Natural Products for Cancer Stem Cells vol.8, 2020, https://doi.org/10.3389/fcell.2020.00283
  2. Pharmacological Inhibition of O-GlcNAc Transferase Promotes mTOR-Dependent Autophagy in Rat Cortical Neurons vol.10, pp.12, 2020, https://doi.org/10.3390/brainsci10120958
  3. Potential Therapeutic Role of Phytochemicals to Mitigate Mitochondrial Dysfunctions in Alzheimer’s Disease vol.10, pp.1, 2020, https://doi.org/10.3390/antiox10010023
  4. Modulatory Effects of Autophagy on APP Processing as a Potential Treatment Target for Alzheimer’s Disease vol.9, pp.1, 2021, https://doi.org/10.3390/biomedicines9010005
  5. Phytochemicals as a Complement to Cancer Chemotherapy: Pharmacological Modulation of the Autophagy-Apoptosis Pathway vol.12, 2021, https://doi.org/10.3389/fphar.2021.639628
  6. Ginseng gintonin, aging societies, and geriatric brain diseases vol.10, pp.1, 2021, https://doi.org/10.1016/j.imr.2020.100450
  7. Antioxidant Compound, Oxyresveratrol, Inhibits APP Production through the AMPK/ULK1/mTOR-Mediated Autophagy Pathway in Mouse Cortical Astrocytes vol.10, pp.3, 2021, https://doi.org/10.3390/antiox10030408
  8. 4,6′-Anhydrooxysporidinone from Fusarium lateritium SSF2 Induces Autophagic and Apoptosis Cell Death in MCF-7 Breast Cancer Cells vol.11, pp.6, 2020, https://doi.org/10.3390/biom11060869
  9. Antioxidant and Anti-Inflammatory Effects of 3-Dehydroxyceanothetric Acid 2-Methyl Ester Isolated from Ziziphus jujuba Mill. against Cisplatin-Induced Kidney Epithelial Cell Death vol.11, pp.11, 2021, https://doi.org/10.3390/biom11111614