The protective effect of epigallocatechin 3-gallate on mouse pancreatic islets via the Nrf2 pathway

  • Yuma Wada
  • Atsushi Takata
  • Tetsuya IkemotoEmail author
  • Yuji Morine
  • Satoru Imura
  • Shuichi Iwahashi
  • Yu Saito
  • Mitsuo Shimada
Original Article



Epigallocatechin 3-gallate (EGCG), a green tea polyphenol, has been shown to have anti-oxidant and anti-inflammatory effects in vitro and in vivo. The aim of this study was to investigate the effects and mechanism of EGCG on isolated pancreatic islets as pre-conditioning for pancreatic islet transplantation.


The pancreatic islets were divided into two groups: an islet culture medium group (control) and an islet culture medium with EGCG (100 µM) group. We investigated the islet viability, Nrf2 expression, reactive oxygen species (ROS) production, and heme oxygenase-1 (HO-1) mRNA. Five hundred islet equivalents after 12 h of culture for the EGCG 100 µM and control group were transplanted under the kidney capsule of streptozotocin-induced diabetic ICR mice.


The cell viability and insulin secretion ability in the EGCG group were preserved, and the nuclear translocation of Nrf2 was increased in the EGCG group (p < 0.01). While the HO-1 mRNA levels were also higher in the EGCG group than in the control group (p < 0.05), the ROS production was lower (p < 0.01). An in vivo functional assessment showed that the blood glucose level had decreased in the EGCG group after transplantation (p < 0.01).


EGCG protects the viability and function of islets by suppressing ROS production via the Nrf2 pathway.


Epigallocatechin 3-gallate (EGCG) Nuclear factor erythroid 2-related factor 2 (Nrf2) Islet transplantation Reactive oxygen species (ROS) production Pre-conditioning 


Compliance with ethical standards

Conflict of interest

All authors have no conflicts of interest.


  1. 1.
    Ryan EA, Paty BW, Senior PA, Bigam D, Alfadhli E, Kneteman NM, et al. Five-year follow-up after clinical islet transplantation. Diabetes. 2005;54:2060–9.CrossRefGoogle Scholar
  2. 2.
    Mandel TE, Koulmanda M. Effect of ischemia and temperature on fetal mouse pancreas. Insulin production in vitro, and function after isotransplantation. Diabetes. 1984;33:376–82.CrossRefGoogle Scholar
  3. 3.
    Sarvetnick N, Shizuru J, Liggitt D, Martin L, McIntyre B, Gregory A, et al. Loss of pancreatic islet tolerance induced by beta-cell expression of interferon-gamma. Nature. 1990;346:844–7.CrossRefGoogle Scholar
  4. 4.
    Kessler L, Jesser C, Lombard Y, Karsten V, Belcourt A, Pinget M, et al. Cytotoxicity of peritoneal murine macrophages against encapsulated pancreatic rat islets: in vivo and in vitro studies. J Leukoc Biol. 1996;60:729–36.CrossRefGoogle Scholar
  5. 5.
    Kumar NB, Pow-Sang J, Spiess PE, Park J, Salup R, Williams CR, Parnes H, Schell MJ. Randomized, placebo-controlled trial evaluating the safety of one-year administration of green tea catechins. Oncotarget. 2016;43:70794–802.Google Scholar
  6. 6.
    Meshitsuka S, Shingaki S, Hotta M, Goto M, Kobayashi M, Ukawa Y, Sagesaka YM, Wada Y, Nojima M, Suzuki K. Phase 2 trial of daily, oral epigallocatechin gallate in patients with light-chain amyloidosis. Int J Hematol. 2017;105:295–308.CrossRefGoogle Scholar
  7. 7.
    Chen IJ, Liu CY, Chiu JP, Hsu CH. Therapeutic effect of high-dose green tea extract on weight reduction: a randomized, double-blind, placebo-controlled clinical trial. Clin Nutr. 2016;35:592–9.CrossRefGoogle Scholar
  8. 8.
    Saito Y, Mori H, Takasu C, Komatsu M, Hanaoka J, Yamada S, et al. Beneficial effects of green tea catechin on massive hepatectomy model in rats. J Gastroenterol. 2014;49:692–701.CrossRefGoogle Scholar
  9. 9.
    Anderson RF, Fisher LJ, Hara Y, Harris T, Mak WB, Melton LD, et al. Green tea catechins partially protect DNA from (.)OH radical-induced strand breaks and base damage through fast chemical repair of DNA radicals. Carcinogenesis. 2001;22:1189–93.CrossRefGoogle Scholar
  10. 10.
    Chow HH, Cai Y, Alberts DS, Hakim I, Dorr R, Shahi F, et al. Phase I pharmacokinetic study of tea polyphenols following single-dose administration of epigallocatechin gallate and polyphenon E. Cancer Epidemiol Biomark Prev. 2001;10:53–8.Google Scholar
  11. 11.
    Tipoe GL, Leung TM, Liong EC, Lau TY, Fung ML, Nanji AA, et al. Epigallocatechin 3-gallate (EGCG) reduces liver inflammation, oxidative stress and fibrosis in carbon tetrachloride (CCl4)-induced liver injury in mice. Toxicology. 2010;273:45–52.CrossRefGoogle Scholar
  12. 12.
    Higdon JV, Frei B. Tea catechins and polyphenols: health effects, metabolism, and antioxidant functions. Crit Rev Food Sci Nutr. 2003;43:89–143.CrossRefGoogle Scholar
  13. 13.
    Kuo PL, Lin CC. Green tea constituent (−)-epigallocatechin 3-gallate inhibits Hep G2 cell proliferation and induces apoptosis through p53-dependent and Fas-mediated pathways. J Biomed Sci. 2003;10:219–27.PubMedGoogle Scholar
  14. 14.
    Surh YJ, Kundu JK, Na HK, Lee JS. Redox-sensitive transcription factors as prime targets for chemoprevention with anti-inflammatory and antioxidative phytochemicals. J Nutr. 2005;135:2993S–3001S.CrossRefGoogle Scholar
  15. 15.
    Shin JH, Jeon HJ, Park J, Chang MS. Epigallocatechin 3-gallate prevents oxidative stress-induced cellular senescence in human mesenchymal stem cells via Nrf2. Int J Mol Med. 2016;38:1075–82.CrossRefGoogle Scholar
  16. 16.
    Yagishita Y, Fukutomi T, Sugawara A, Kawamura H, Takahashi T, Pi J, et al. Nrf2 protects pancreatic beta-cells from oxidative and nitrosative stress in diabetic model mice. Diabetes. 2014;63:605–18.CrossRefGoogle Scholar
  17. 17.
    Itoh K, Wakabayashi N, Katoh Y, Ishii T, Igarashi K, Engel JD, et al. Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev. 1999;13:76–86.CrossRefGoogle Scholar
  18. 18.
    Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem. 2009;284:13291–5.CrossRefGoogle Scholar
  19. 19.
    Sahin K, Tuzcu M, Gencoglu H, Dogukan A, Timurkan M, Sahin N, et al. Epigallocatechin 3-gallate activates Nrf2/HO-1 signaling pathway in cisplatin induced nephrotoxicity in rats. Life Sci. 2010;87:240–5.CrossRefGoogle Scholar
  20. 20.
    Pang X, Xue W, Feng X, Tian X, Teng Y, Ding X, et al. Experimental studies on islets isolation, purification and function in rats. Int J Clin Exp Med. 2015;8:20932–8.PubMedPubMedCentralGoogle Scholar
  21. 21.
    Li S, Vaziri ND, Masuda Y, Hajighasemi-Ossareh M, Robles L, Le A, et al. Pharmacological activation of Nrf2 pathway improves pancreatic islet isolation and transplantation. Cell Transplant. 2015;24:2273–83.CrossRefGoogle Scholar
  22. 22.
    Rabinovitch A, Sorensen O, Suarez-Pinzon WL, Power RF, Rajotte RV, Bleackley RC, et al. Analysis of cytokine mRNA expression in syngeneic islet grafts of NOD mice: interleukin 2 and interferon gamma mRNA expression correlate with graft rejection and interleukin 10 with graft survival. Diabetologia. 1994;37:833–7.CrossRefGoogle Scholar
  23. 23.
    Faust A, Rothe H, Schade U, Lampeter E, Kolb H, et al. Primary nonfunction of islet grafts in autoimmune diabetic nonobese diabetic mice is prevented by treatment with interleukin 4 and interleukin 10. Transplantation. 1996;62:648–52.CrossRefGoogle Scholar
  24. 24.
    Dobson T, Fraga D, Saba C, Bryer-Ash M, Gaber AO, Gerling IC, et al. Human pancreatic islets transfected to produce an inhibitor of TNF are protected against destruction by human leukocytes. Cell Transplant. 2000;9:857–65.CrossRefGoogle Scholar
  25. 25.
    Hennige AM, Lembert N, Wahl MA, Ammon HP. Oxidative stress increases potassium efflux from pancreatic islets by depletion of intracellular calcium stores. Free Radic Res. 2000;33:507–16.CrossRefGoogle Scholar
  26. 26.
    Takahashi T, Shimizu H, Morimatsu H, Maeshima K, Inoue K, Akagi R, et al. Heme oxygenase-1 is an essential cytoprotective component in oxidative tissue injury induced by hemorrhagic shock. J Clin Biochem Nutr. 2009;44:28–40.CrossRefGoogle Scholar
  27. 27.
    Bird JE, Giancarli MR, Megill JR, Durham SK. Effects of endothelin in radiocontrast-induced nephropathy in rats are mediated through endothelin-A receptors. J Am Soc Nephrol. 1996;7:1153–7.PubMedGoogle Scholar
  28. 28.
    Luo YP, Jiang L, Kang K, Fei DS, Meng XL, Nan CC, et al. Hemin inhibits NLRP3 inflammasome activation in sepsis-induced acute lung injury, involving heme oxygenase-1. Int Immunopharmacol. 2014;20:24–32.CrossRefGoogle Scholar
  29. 29.
    Kim SJ, Lee SM. NLRP3 inflammasome activation in D-galactosamine and lipopolysaccharide-induced acute liver failure: role of heme oxygenase-1. Free Radic Biol Med. 2013;65:997–1004.CrossRefGoogle Scholar
  30. 30.
    Scapagnini G, Vasto S, Abraham NG, Caruso C, Zella D, Fabio G, et al. Modulation of Nrf2/ARE pathway by food polyphenols: a nutritional neuroprotective strategy for cognitive and neurodegenerative disorders. Mol Neurobiol. 2011;44:192–201.CrossRefGoogle Scholar
  31. 31.
    Pan H, Chen J, Shen K, Wang X, Wang P, Fu G, et al. Mitochondrial modulation by Epigallocatechin 3-Gallate ameliorates cisplatin induced renal injury through decreasing oxidative/nitrative stress, inflammation and NF-kB in mice. PLoS One. 2015;10:e0124775.CrossRefGoogle Scholar
  32. 32.
    Kakuta Y, Okumi M, Isaka Y, Tsutahara K, Abe T, Yazawa K, et al. Epigallocatechin 3-gallate protects kidneys from ischemia reperfusion injury by HO-1 upregulation and inhibition of macrophage infiltration. Transpl Int. 2011;24:514–22.CrossRefGoogle Scholar
  33. 33.
    Ellrichmann G, Petrasch-Parwez E, Lee DH, Reick C, Arning L, Saft C, et al. Efficacy of fumaric acid esters in the R6/2 and YAC128 models of Huntington’s disease. PLoS One. 2011;6:e16172.CrossRefGoogle Scholar
  34. 34.
    Xu J, Huang G, Zhang K, Sun J, Xu T, Li R, et al. Nrf2 activation in astrocytes contributes to spinal cord ischemic tolerance induced by hyperbaric oxygen preconditioning. J Neurotrauma. 2014;31:1343–53.CrossRefGoogle Scholar
  35. 35.
    Okada K, Shoda J, Kano M, Suzuki S, Ohtake N, Yamamoto M, et al. Inchinkoto, a herbal medicine, and its ingredients dually exert Mrp2/MRP2-mediated choleresis and Nrf2-mediated antioxidative action in rat livers. Am J Physiol Gastrointest Liver Physiol. 2007;292:G1450-63.CrossRefGoogle Scholar
  36. 36.
    Makabe S, Takahashi Y, Watanabe H, Murakami M, Ohba T, Ito H, et al. Fluvastatin protects vascular smooth muscle cells against oxidative stress through the Nrf2-dependent antioxidant pathway. Atherosclerosis. 2010;213:377–84.CrossRefGoogle Scholar
  37. 37.
    de Zeeuw D, Akizawa T, Audhya P, Bakris GL, Chin M, Christ-Schmidt H, et al. Bardoxolone methyl in type 2 diabetes and stage 4 chronic kidney disease. N Engl J Med. 2013;369:2492–503.CrossRefGoogle Scholar
  38. 38.
    Nakai K, Fujii H, Kono K, Goto S, Kitazawa R, Kitazawa S, et al. Vitamin D activates the Nrf2-Keap1 antioxidant pathway and ameliorates nephropathy in diabetic rats. Am J Hypertens. 2014;27:586–95.CrossRefGoogle Scholar
  39. 39.
    Rubiolo JA, Mithieux G, Vega FV. Resveratrol protects primary rat hepatocytes against oxidative stress damage: activation of the Nrf2 transcription factor and augmented activities of antioxidant enzymes. Eur J Pharmacol. 2008;591:66–72.CrossRefGoogle Scholar
  40. 40.
    Morimitsu Y, Nakagawa Y, Hayashi K, Fujii H, Kumagai T, Nakamura Y, et al. A sulforaphane analogue that potently activates the Nrf2-dependent detoxification pathway. J Biol Chem. 2002;277:3456–63.CrossRefGoogle Scholar
  41. 41.
    Yagi H, Tan J, Tuan RS. Polyphenols suppress hydrogen peroxide-induced oxidative stress in human bone-marrow derived mesenchymal stem cells. J Cell Biochem. 2013;114:1163–73.CrossRefGoogle Scholar
  42. 42.
    Grube S, Ewald C, Kögler C, Lawson McLean A, Kalff R, Walter J. Achievable central nervous system concentrations of the green tea catechin EGCG induce stress in glioblastoma cells in vitro. Nutr Cancer. 2018;10:1–14.CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  1. 1.Department of Surgery, Graduate School of Biomedical SciencesTokushima UniversityTokushimaJapan

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