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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

Abstract

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.

Keywords

Senescence MTOR Senolytic ROS EGCG 

Abbreviations

EGCG

Epigallocatechin gallate

mTOR

Mechanistic target of rapamycin

H2O2

Hydrogen peroxide

SASP

Senescence-associated secretory phenotype

ROS

Reactive oxygen species

iNOS

Inducible nitric oxide synthase

Cox-2

Cyclooxygenase-2

NF-κB

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

AMPK

Adenosine monophosphate-activated protein kinase

Notes

Acknowledgements

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)

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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

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