Protective effects of 6,7,4′-trihydroxyisoflavone, a major metabolite of daidzein, on 6-hydroxydopamine-induced neuronal cell death in SH-SY5Y human neuroblastoma cells

  • Yong-Hyun Ko
  • Seung-Hwan Kwon
  • Seon-Kyung Kim
  • Bo-Ram Lee
  • Kwang-Hyun Hur
  • Young-Jung Kim
  • Seong-Eon Kim
  • Seok-Yong Lee
  • Choon-Gon JangEmail author
Research Article


Daidzein, one of the important isoflavones, is extensively metabolized in the human body following consumption. In particular, 6,7,4′-trihydroxyisoflavone (THIF), a major metabolite of daidzein, has been the focus of recent investigations due to its various health benefits, such as anti-cancer and anti-obesity effects. However, the protective effects of 6,7,4′-THIF have not yet been studied in models of Parkinson’s disease (PD). Therefore, the present study aimed to investigate the protective activity of 6,7,4′-THIF on 6-hydroxydopamine (OHDA)-induced neurotoxicity in SH-SY5Y human neuroblastoma cells. Pretreatment of SH-SY5Y cells with 6,7,4′-THIF significantly inhibited 6-OHDA-induced neuronal cell death, lactate dehydrogenase release, and reactive oxygen species production. In addition, 6,7,4′-THIF significantly attenuated reductions in 6-OHDA-induced superoxide dismutase activity and glutathione content. Moreover, 6,7,4′-THIF attenuated alterations in Bax and Bcl-2 expression and caspase-3 activity in 6-OHDA-induced SH-SY5Y cells. Furthermore, 6,7,4′-THIF significantly reduced 6-OHDA-induced phosphorylation of c-Jun N-terminal kinase, p38 mitogen-activated protein kinase, and extracellular signal-regulated kinase 1/2. Additionally, 6,7,4′-THIF effectively prevented 6-OHDA-induced loss of tyrosine hydroxylase. Taken together, these results suggest that 6,7,4′-THIF, a major metabolite of daidzein, may be an attractive option for treating and/or preventing neurodegenerative disorders such as PD.


6,7,4′-Trihydroxyisoflavone 6-Hydroxydopamine Oxidative stress Apoptosis Parkinson’s disease 



This work was supported by a Grant (NRF-2012R1A5A2A28671860) from the Basic Science Research Program through the National Research Foundation of Korea.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare.


  1. Atherton KM, Mutch E, Ford D (2006) Metabolism of the soyabean isoflavone daidzein by CYP1A2 and the extra-hepatic CYPs 1A1 and 1B1 affects biological activity. Biochem Pharmacol 72:624–631. CrossRefPubMedGoogle Scholar
  2. Blum D, Torch S, Lambeng N, Nissou M, Benabid AL, Sadoul R, Verna JM (2001) Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson’s disease. Prog Neurobiol 65:135–172. CrossRefPubMedGoogle Scholar
  3. Cardoso SM, Moreira PI, Agostinho P, Pereira C, Oliveira CR (2005) Neurodegenerative pathways in Parkinson’s disease: therapeutic strategies. Curr Drug Targets CNS Neurol Disord 4:405–419. CrossRefPubMedGoogle Scholar
  4. Chen JH, Ou HP, Lin CY, Lin FJ, Wu CR, Chang SW, Tsai CW (2012) Carnosic acid prevents 6-hydroxydopamine-induced cell death in SH-SY5Y cells via mediation of glutathione synthesis. Chem Res Toxicol 25:1893–1901. CrossRefPubMedGoogle Scholar
  5. Chi SW (2014) Structural insights into the transcription-independent apoptotic pathway of p53. BMB Rep 47:167–172. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Choi WS, Eom DS, Han BS, Kim WK, Han BH, Choi EJ, Oh TH, Markelonis GJ, Cho JW, Oh YJ (2004) Phosphorylation of p38 MAPK induced by oxidative stress is linked to activation of both caspase-8- and -9-mediated apoptotic pathways in dopaminergic neurons. J Biol Chem 279:20451–20460. CrossRefPubMedGoogle Scholar
  7. Cunha MP, Martín-de-Saavedra MD, Romero A, Parada E, Egea J, Del Barrio L, Rodrigues AL, López MG (2013) Protective effect of creatine against 6-hydroxydopamine-induced cell death in human neuroblastoma SH-SY5Y cells: involvement of intracellular signaling pathways. Neuroscience 238:185–194. CrossRefPubMedGoogle Scholar
  8. Di Carlo G, Mascolo N, Izzo AA, Capasso F (1999) Flavonoids: old and new aspects of a class of natural therapeutic drugs. Life Sci 65:337–353. CrossRefPubMedGoogle Scholar
  9. Dzamko N, Zhou J, Huang Y, Halliday GM (2014) Parkinson’s disease-implicated kinases in the brain; insights into disease pathogenesis. Front Mol Neurosci 7:57. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Franco JL, Posser T, Gordon SL, Bobrovskaya L, Schneider JJ, Farina M, Dafre AL, Dickson PW, Dunkley PR (2010) Expression of tyrosine hydroxylase increases the resistance of human neuroblastoma cells to oxidative insults. Toxicol Sci 113:150–157. CrossRefPubMedGoogle Scholar
  11. Harper SJ, Wilkie N (2003) MAPKs: new targets for neurodegeneration. Expert Opin Ther Targets 7:187–200. CrossRefPubMedGoogle Scholar
  12. Hirota A, Inaba M, Chen YC, Abe N, Taki S, Yano M, Kawaii S (2004) Isolation of 8-hydroxyglycitein and 6-hydroxydaidzein from soybean miso. Biosci Biotechnol Biochem 68:1372–1374. CrossRefPubMedGoogle Scholar
  13. Kirik D, Rosenblad C, Burger C, Lundberg C, Johansen TE, Muzyczka N, Mandel RJ, Björklund A (2002) Parkinson-like neurodegeneration induced by targeted overexpression of alpha-synuclein in the nigrostriatal system. J Neurosci 22:2780–2791. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Ko YH, Kim SY, Lee SY, Jang CG (2018a) 6,7,4′-Trihydroxyisoflavone, a major metabolite of daidzein, improves learning and memory via the cholinergic system and the p-CREB/BDNF signaling pathway in mice. Eur J Pharmacol 826:140–147. CrossRefPubMedGoogle Scholar
  15. Ko YH, Kwon SH, Ma SX, Seo JY, Lee BR, Kim K, Kim SY, Lee SY, Jang CG (2018b) The memory-enhancing effects of 7,8,4′-trihydroxyisoflavone, a major metabolite of daidzein, are associated with activation of the cholinergic system and BDNF signaling pathway in mice. Brain Res Bull 142:197–206. CrossRefPubMedGoogle Scholar
  16. Kwon SH, Kim JA, Hong SI, Jung YH, Kim HC, Lee SY, Jang CG (2011) Loganin protects against hydrogen peroxide-induced apoptosis by inhibiting phosphorylation of JNK, p38, and ERK 1/2 MAPKs in SH-SY5Y cells. Neurochem Int 58:533–541. CrossRefPubMedGoogle Scholar
  17. Kwon SH, Ma SX, Lee SY, Jang CG (2014) Sulfuretin inhibits 6-hydroxydopamine-induced neuronal cell death via reactive oxygen species-dependent mechanisms in human neuroblastoma SH-SY5Y cells. Neurochem Int 74:53–64. CrossRefPubMedGoogle Scholar
  18. Kwon SH, Hong SI, Ma SX, Lee SY, Jang CG (2015) 3′,4′,7-Trihydroxyflavone prevents apoptotic cell death in neuronal cells from hydrogen peroxide-induced oxidative stress. Food Chem Toxicol 80:41–51. CrossRefPubMedGoogle Scholar
  19. Lee DE, Lee KW, Jung SK, Lee EJ, Hwang JA, Lim TG, Kim BY, Bode AM, Lee HJ, Dong Z (2011) 6,7,4′-Trihydroxyisoflavone inhibits HCT-116 human colon cancer cell proliferation by targeting CDK1 and CDK2. Carcinogenesis 32:629–632. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Lewis TS, Shapiro PS, Ahn NG (1998) Signal transduction through MAP kinase cascades. Adv Cancer Res 74:49–139. CrossRefPubMedGoogle Scholar
  21. Li T, Feng Y, Yang R, Wu L, Li R, Huang L, Yang Q, Chen J (2018) Salidroside promotes the pathological α-synuclein clearance through ubiquitin-proteasome system in SH-SY5Y cells. Front Pharmacol 9:377. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Lim TG, Lee SY, Duan Z, Lee MH, Chen H, Liu F, Liu K, Jung SK, Kim DJ, Bode AM, Lee KW, Dong Z (2017) The prolyl isomerase PIN1 is a novel target of 6,7,4′-trihydroxyisoflavone for suppressing esophageal cancer growth. Cancer Prev Res (Phila) 10:308–318. CrossRefGoogle Scholar
  23. Lin CM, Lin RD, Chen ST, Lin YP, Chiu WT, Lin JW, Hsu FL, Lee MH (2010) Neurocytoprotective effects of the bioactive constituents of Pueraria thomsonii in 6-hydroxydopamine (6-OHDA)-treated nerve growth factor (NGF)-differentiated PC12 cells. Phytochemistry 71:2147–2156. CrossRefPubMedGoogle Scholar
  24. Liu H, Mao P, Wang J, Wang T, Xie CH (2015) Allicin protects PC12 cells against 6-OHDA-induced oxidative stress and mitochondrial dysfunction via regulating mitochondrial dynamics. Cell Physiol Biochem 36:966–979. CrossRefPubMedGoogle Scholar
  25. Lotharius J, Dugan LL, O’Malley KL (1999) Distinct mechanisms underlie neurotoxin-mediated cell death in cultured dopaminergic neurons. J Neurosci 19:1284–1293. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Lou H, Jing X, Wei X, Shi H, Ren D, Zhang X (2014) Naringenin protects against 6-OHDA-induced neurotoxicity via activation of the Nrf2/ARE signaling pathway. Neuropharmacology 79:380–388. CrossRefPubMedGoogle Scholar
  27. Mao QQ, Xian YF, Ip SP, Tsai SH, Che CT (2011) Protective effects of peony glycosides against corticosterone-induced cell death in PC12 cells through antioxidant action. J Ethnopharmacol 133:1121–1125. CrossRefPubMedGoogle Scholar
  28. Meng H, Fu G, Shen J, Shen K, Xu Z, Wang Y, Jin B, Pan H (2017) Ameliorative effect of daidzein on cisplatin-induced nephrotoxicity in mice via modulation of inflammation, oxidative stress, and cell death. Oxid Med Cell Longev 2017:3140680. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Mueller M, Hobiger S, Jungbauer A (2010) Red clover extract: a source for substances that activate peroxisome proliferator-activated receptor alpha and ameliorate the cytokine secretion profile of lipopolysaccharide-stimulated macrophages. Menopause 17:379–387. CrossRefPubMedGoogle Scholar
  30. Park MH, Ju JW, Kim M, Han JS (2016) The protective effect of daidzein on high glucose-induced oxidative stress in human umbilical vein endothelial cells. Z Naturforsch C 71:21–28. CrossRefPubMedGoogle Scholar
  31. Park HJ, Lee KS, Zhao TT, Lee KE, Lee MK (2017) Effects of asarinin on dopamine biosynthesis and 6-hydroxydopamine-induced cytotoxicity in PC12 cells. Arch Pharm Res 40:631–639. CrossRefPubMedGoogle Scholar
  32. Pišlar AH, Zidar N, Kikelj D, Kos J (2014) Cathepsin X promotes 6-hydroxydopamine-induced apoptosis of PC12 and SH-SY5Y cells. Neuropharmacology 82:121–131. CrossRefPubMedGoogle Scholar
  33. Silva J, Alves C, Pinteus S, Mendes S, Pedrosa R (2018) Neuroprotective effects of seaweeds against 6-hydroxidopamine-induced cell death on an in vitro human neuroblastoma model. BMC Complement Altern Med 18:58. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Snyder H, Mensah K, Hsu C, Hashimoto M, Surgucheva IG, Festoff B, Surguchov A, Masliah E, Matouschek A, Wolozin B (2005) Beta-synuclein reduces proteasomal inhibition by alpha-synuclein but not gamma-synuclein. J Biol Chem 280:7562–7569. CrossRefPubMedGoogle Scholar
  35. Song JX, Shaw PC, Sze CW, Tong Y, Yao XS, Ng TB, Zhang YB (2010) Chrysotoxine, a novel bibenzyl compound, inhibits 6-hydroxydopamine induced apoptosis in SH-SY5Y cells via mitochondria protection and NF-κB modulation. Neurochem Int 57:676–689. CrossRefPubMedGoogle Scholar
  36. Tan JW, Kim MK (2016) Neuroprotective effects of biochanin A against β-amyloid-induced neurotoxicity in PC12 cells via a mitochondrial-dependent apoptosis pathway. Molecules 21:548. CrossRefPubMedCentralGoogle Scholar
  37. Tortosa A, López E, Ferrer I (1997) Bcl-2 and Bax proteins in Lewy bodies from patients with Parkinson’s disease and diffuse Lewy body disease. Neurosci Lett 238:78–80. CrossRefPubMedGoogle Scholar
  38. Vander Heiden MG, Thompson CB (1999) Bcl-2 proteins: regulators of apoptosis or of mitochondrial homeostasis? Nat Cell Biol 1:E209–E216. CrossRefPubMedGoogle Scholar
  39. Xiao X, Liu J, Hu J, Zhu X, Yang H, Wang C, Zhang Y (2008) Protective effects of protopine on hydrogen peroxide-induced oxidative injury of PC12 cells via Ca(2+) antagonism and antioxidant mechanisms. Eur J Pharmacol 591:21–27. CrossRefPubMedGoogle Scholar
  40. Xie HR, Hu LS, Li GY (2010) SH-SY5Y human neuroblastoma cell line: in vitro cell model of dopaminergic neurons in Parkinson’s disease. Chin Med J (Engl) 123:1086–1092. CrossRefGoogle Scholar
  41. Yuan WJ, Yasuhara T, Shingo T, Muraoka K, Agari T, Kameda M, Uozumi T, Tajiri N, Morimoto T, Jing M, Baba T, Wang F, Leung H, Matsui T, Miyoshi Y, Date I (2008) Neuroprotective effects of edaravone-administration on 6-OHDA-treated dopaminergic neurons. BMC Neurosci 9:75. CrossRefPubMedGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2019

Authors and Affiliations

  • Yong-Hyun Ko
    • 1
  • Seung-Hwan Kwon
    • 1
  • Seon-Kyung Kim
    • 1
  • Bo-Ram Lee
    • 1
  • Kwang-Hyun Hur
    • 1
  • Young-Jung Kim
    • 1
  • Seong-Eon Kim
    • 1
  • Seok-Yong Lee
    • 1
  • Choon-Gon Jang
    • 1
    Email author
  1. 1.Department of Pharmacology, School of PharmacySungkyunkwan UniversitySuwonRepublic of Korea

Personalised recommendations