Research Paper Volume 11, Issue 11 pp 3731—3749

Nectandrin B-mediated activation of the AMPK pathway prevents cellular senescence in human diploid fibroblasts by reducing intracellular ROS levels

Hyun-Jin Jang1,2, *, , Kyeong Eun Yang1, *, , Won Keun Oh3, *, , Song-I Lee4, , In-Hu Hwang5, , Kyung-Tae Ban6, , Hwa-Seung Yoo6, , Jong-Soon Choi1, , Eui-Ju Yeo4,7, , Ik-Soon Jang1,8, ,

  • 1 Drug & Disease Target Group, Division of Bioconvergence Analysis, Korea Basic Science Institute, Daejeon 305-333, Republic of Korea
  • 2 Department of Biological Sciences, Sungkyunkwan University, Suwon 16419, Republic of Korea
  • 3 Korea Bioactive Natural Material Bank, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea
  • 4 Department of Health Sciences and Technology, GAIHST, Gachon University, Incheon 21999, Republic of Korea
  • 5 Neuroscience Research Institute, Korea University College of Medicine, Seoul 136-705, Republic of Korea
  • 6 East-West Cancer Center, Daejeon University, Daejeon, 302-120, Republic of Korea
  • 7 Department of Biochemistry, College of Medicine, Gachon University, Incheon 21999, Republic of Korea
  • 8 Division of Analytical Science, University of Science and Technology, Daejeon 34113, Republic of Korea
* Equal contribution

Received: March 12, 2019       Accepted: May 31, 2019       Published: June 14, 2019      

https://doi.org/10.18632/aging.102013
How to Cite

Copyright: Jang et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Nectandrin B (NecB) is a bioactive lignan compound isolated from Myristica fragrans (nutmeg), which functions as an activator of AMP-activated protein kinase (AMPK). Because we recently found that treatment with NecB increased the cell viability of old human diploid fibroblasts (HDFs), the underlying molecular mechanism was investigated. NecB treatment in old HDFs reduced the activity staining of senescence-associated β-galactosidase and the levels of senescence markers, such as the Ser15 phosphorylated p53, caveolin-1, p21waf1, p16ink4a, p27kip1, and cyclin D1. NecB treatment increased that in S phase, indicating a enhancement of cell cycle entry. Interestingly, NecB treatment ameliorated age-dependent activation of AMPK in old HDFs. Moreover, NecB reversed the age-dependent expression and/or activity changes of certain sirtuins (SIRT1−5), and cell survival/death-related proteins. The transcriptional activity of Yin-Yang 1 and the expression of downstream proteins were elevated in NecB-treated old HDFs. In addition, NecB treatment exerted a radical scavenging effect in vitro, reduced cellular ROS levels, and increased antioxidant enzymes in old HDFs. Moreover, NecB-mediated activation of the AMPK pathway reduced intracellular ROS levels. These results suggest that NecB-induced protection against cellular senescence is mediated by ROS-scavenging through activation of AMPK. NecB might be useful in ameliorating age-related diseases and extending human lifespan.

Abbreviations

AMPK: AMP-activated protein kinase; DMEM: Dulbecco’s modified Eagle’s medium; ECL: enhanced chemiluminescence; ERK: extracellular signal-regulated kinase; FBS: fetal bovine serum; HDFs: human diploid fibroblasts; MTT: 3-(4,5-dimethylthiazol-e-yl)-2,5-diphenyltetrazolium bromide; NAC: N-acetylcysteine; NecB: nectandrin B; PEG: polyethylene glycol-300; PI: propidium iodide; ROS: reactive oxygen species; SA-β-gal: senescence-associated β-galactosidase; SIRT: sirtuin NAD-dependent deacetylase; X-gal: 5-bromo-4-chloro-3-indolyl-b-d-galactopyranoside.