Epigallocatechin gallate suppresses premature senescence of preadipocytes by inhibition of PI3K/Akt/mTOR pathway and induces senescent cell death by regulation of Bax/Bcl-2 pathway

  • Ravi Kumar
  • Anamika Sharma
  • Amita Kumari
  • Ashu Gulati
  • Yogendra PadwadEmail author
  • Rohit SharmaEmail author
Research Article


The phytochemical epigallocatechin gallate (EGCG) has been reported to alleviate age-associated immune disorders and organ dysfunction. However, information regarding the mechanistic role of EGCG in the suppression of cellular senescence is limited. The present study thus assessed the effects and underlying mechanisms of EGCG in the inhibition of senescence as well as its potential to selectively eliminate senescent cells (senolytics) using 3T3-L1 preadipocytes. Premature senescence was established in cells by repeated exposure of H2O2 at a sub-lethal concentration (150 μM). H2O2 treated cells showed characteristic senescence-associated features including increased cell size, senescence-associated β-galactosidase activity (SA-β-gal), development of senescence-associated secretory phenotype (SASP), activation of reactive oxygen species (ROS) and pathways, DNA damage as well as induction of cell cycle inhibitors (p53/p21WAF1/p16INK4a). In addition, a robust activation of PI3K/Akt/mTOR and AMPK pathways was also observed in H2O2 treated cells. Presence of EGCG (50 and 100 μM) showed significant downregulation of PI3K/Akt/mTOR and AMPK signaling along with the suppression of ROS, iNOS, Cox-2, NF-κB, SASP and p53 mediated cell cycle inhibition in preadipocytes. In addition, EGCG treatment also suppressed the accumulation of anti-apoptotic protein Bcl-2 in senescent cells thereby promoting apoptosis mediated cell death. Our results collectively show that EGCG acts as an mTOR inhibitor, SASP modulator as well as a potential senolytic agent thereby indicating its multi-faceted attributes that could be useful for developing anti-aging or age-delaying therapies.


Senescence MTOR Senolytic ROS EGCG 



Epigallocatechin gallate


Mechanistic target of rapamycin


Hydrogen peroxide


Senescence-associated secretory phenotype


Reactive oxygen species


Inducible nitric oxide synthase




Nuclear factor kappa-light-chain-enhancer of activated B cells


Adenosine monophosphate-activated protein kinase



Authors are grateful to the Director, CSIR-IHBT for constant encouragement and support. This work was supported by grants from Department of Science and Technology, Government of India under the INSPIRE Faculty scheme (IFA17-LSPA79) and CSIR in-house project MLP0204.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

10522_2018_9785_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 14 kb)
10522_2018_9785_MOESM2_ESM.docx (60 kb)
Supplementary material 2 (DOCX 60 kb)


  1. Bent EH, Gilbert LA, Hemann MT (2016) A senescence secretory switch mediated by PI3K/AKT/mTOR activation controls chemoprotective endothelial secretory responses. Genes Dev 30(16):1811–1821CrossRefGoogle Scholar
  2. Bhatia-Dey N, Kanherkar RR, Stair SE, Makarev EO, Csoka AB (2016) Cellular senescence as the causal nexus of aging. Front Genet 7:13CrossRefGoogle Scholar
  3. Biran A, Zada L, Abou Karam P, Vadai E, Roitman L, Ovadya Y, Porat Z, Krizhanovsky V (2017) Quantitative identification of senescent cells in aging and disease. Aging Cell 16(4):661–671CrossRefGoogle Scholar
  4. Blagosklonny MV (2008) Aging: ROS or TOR. Cell Cycle 7(21):3344–3354CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Brown MK, Evans JL, Luo Y (2006) Beneficial effects of natural antioxidants EGCG and alpha-lipoic acid on life span and age-dependent behavioral declines in Caenorhabditis elegans. Pharmacol Biochem Behav 85(3):620–628CrossRefGoogle Scholar
  7. Campisi J (2013) Aging, cellular senescence, and cancer. Annu Rev Physiol 75:685–705CrossRefGoogle Scholar
  8. Carroll B, Korolchuk VI (2017) Dysregulation of mTORC1/autophagy axis in senescence. Aging 9(8):1851–1852PubMedPubMedCentralGoogle Scholar
  9. Chen JH, Ozanne SE, Hales CN (2007) Methods of cellular senescence induction using oxidative stress. Methods Mol Biol 371:179–189CrossRefGoogle Scholar
  10. Choo KB, Tai L, Hymavathee KS, Wong CY, Nguyen PN, Huang CJ, Cheong SK, Kamarul T (2014) Oxidative stress-induced premature senescence in Wharton’s jelly-derived mesenchymal stem cells. Int J Med Sci 11(11):1201–1207CrossRefGoogle Scholar
  11. Chung HY, Lee EK, Choi YJ, Kim JM, Kim DH, Zou Y, Kim CH, Lee J, Kim HS, Kim ND, Jung JH, Yu BP (2011) Molecular inflammation as an underlying mechanism of the aging process and age-related diseases. J Dental Res 90:830–840CrossRefGoogle Scholar
  12. Davalli P, Mitic T, Caporali A, Lauriola A, D’Arca D (2016) ROS, cell senescence, and novel molecular mechanisms in aging and age-related diseases. Oxid Med Cell Longev 2016:3565127CrossRefGoogle Scholar
  13. De la Fuente M, Miquel J (2009) An update of the oxidation-inflammation theory of aging: the involvement of the immune system in oxi-inflamm-aging. Curr Pharm Des 15(26):3003–3026CrossRefGoogle Scholar
  14. Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312CrossRefGoogle Scholar
  15. Han DW, Lee MH, Kim B, Lee JJ, Hyon SH, Park JC (2012) Preventive effects of epigallocatechin-3-O-gallate against replicative senescence associated with p53 acetylation in human dermal fibroblasts. Oxid Med Cell Longev 2012:850684CrossRefGoogle Scholar
  16. Johnson SC, Rabinovitch PS, Kaeberlein M (2013) mTOR is a key modulator of ageing and age-related disease. Nature 493(7432):338–345CrossRefGoogle Scholar
  17. Kennedy BK, Lamming DW (2016) The mechanistic target of rapamycin: the grand conductor of metabolism and aging. Cell Metab 23(6):990–1003CrossRefGoogle Scholar
  18. Kirkland JL, Tchkonia T (2017) Cellular senescence: a translational perspective. EBioMedicine 21:21–28CrossRefGoogle Scholar
  19. Lambert JD, Lee MJ, Lu H, Meng X, Hong JJ, Seril DN, Sturgill MG, Yang CS (2003) Epigallocatechin-3-gallate is absorbed but extensively glucuronidated following oral administration to mice. J Nutr 133(12):4172–4177CrossRefGoogle Scholar
  20. Laplante M, Sabatini DM (2012) mTOR signaling in growth control and disease. Cell 149:274–293CrossRefGoogle Scholar
  21. Mária J, Ingrid Ž (2017) Effects of bioactive compounds on senescence and components of senescence associated secretory phenotypes in vitro. Food Funct 8(7):2394–2418CrossRefGoogle Scholar
  22. Nacarelli T, Azar A, Sell C (2015) Aberrant mTOR activation in senescence and aging: a mitochondrial stress response? Exp Gerontol 68:66–70CrossRefGoogle Scholar
  23. Niu Y, Na L, Feng R, Gong L, Zhao Y, Li Q, Li Y, Sun C (2013) The phytochemical, EGCG, extends lifespan by reducing liver and kidney function damage and improving age-associated inflammation and oxidative stress in healthy rats. Aging Cell 12(6):1041–1049CrossRefGoogle Scholar
  24. Nogueira V, Park Y, Chen CC, Xu PZ, Chen ML, Tonic I, Unterman T, Hay N (2008) Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell 14(6):458–470CrossRefGoogle Scholar
  25. Sadowska-Bartosz I, Bartosz G (2014) Effect of antioxidants supplementation on aging and longevity. Biomed Res Int 2014:404680CrossRefGoogle Scholar
  26. Sharma R, Kapila R, Haq MR, Salingati V, Kapasiya M, Kapila S (2014) Age-associated aberrations in mouse cellular and humoral immune responses. Aging Clin Exp Res 26(4):353–362CrossRefGoogle Scholar
  27. Sharma R, Sharma A, Kumari A, Kulurkar PM, Raj R, Gulati A, Padwad YS (2017) Consumption of green tea epigallocatechin-3-gallate enhances systemic immune response, antioxidative capacity and HPA axis functions in aged male swiss albino mice. Biogerontology 18(3):367–382CrossRefGoogle Scholar
  28. Swami M (2008) Akt: a double-edged sword. Nat Rev Cancer 2008(9):76. CrossRefGoogle Scholar
  29. Trabucco SE, Zhang H (2016) Finding Shangri-La: limiting the impact of senescence on aging. Cell Stem Cell 18(3):305–306CrossRefGoogle Scholar
  30. Verburgh K (2015) Nutrigerontology: why we need a new scientific discipline to develop diets and guidelines to reduce the risk of aging-related diseases. Aging Cell 14(1):17–24CrossRefGoogle Scholar
  31. Wiley CD, Campisi J (2016) From ancient pathways to aging cells-connecting metabolism and cellular senescence. Cell Metab 23(6):1013–1021CrossRefGoogle Scholar
  32. Zhang L, Jie G, Zhang J, Zhao B (2009) Significant longevity-extending effects of EGCG on Caenorhabditis elegans under stress. Free Radic Biol Med 46(3):414–421CrossRefGoogle Scholar
  33. Zhang J, Wang X, Vikash V, Ye Q, Wu D, Liu Y, Dong W (2016) ROS and ROS-mediated cellular signaling. Oxid Med Cell Longev 2016:4350965PubMedPubMedCentralGoogle Scholar
  34. Zhou L, Chen X, Liu T, Gong Y, Chen S, Pan G, Cui W, Luo ZP, Pei M, Yang H, He F (2015) Melatonin reverses H2O2-induced premature senescence in mesenchymal stem cells via the SIRT1-dependent pathway. J Pineal Res 59(2):190–205CrossRefGoogle Scholar
  35. Zhu X, Yue H, Guo X, Yang J, Liu J, Liu J, Wang R, Zhu W (2017) The preconditioning of berberine suppresses hydrogen peroxide-induced premature senescence via regulation of sirtuin 1. Oxid Med Cell Longev 2017:2391820CrossRefGoogle Scholar
  36. Zoico E, Di Francesco V, Olioso D, Fratta Pasini AM, Sepe A, Bosello O, Cinti S, Cominacini L, Zamboni M (2010) In vitro aging of 3T3-L1 mouse adipocytes leads to altered metabolism and response to inflammation. Biogerontology 1:111–122CrossRefGoogle Scholar
  37. Zwerschke W, Mazurek S, Stöckl P, Hütter E, Eigenbrodt E, Jansen-Dürr P (2003) Metabolic analysis of senescent human fibroblasts reveals a role for AMP in cellular senescence. Biochem J 376(Pt 2):403–411CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Pharmacology and Toxicology Laboratory, Food & Nutraceutical DivisionCSIR-Institute of Himalayan Bioresource TechnologyPalampurIndia
  2. 2.Food & Nutraceutical DivisionCSIR-Institute of Himalayan Bioresource TechnologyPalampurIndia

Personalised recommendations