Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Ginsenoside Rb2 suppresses cellular senescence of human dermal fibroblasts by inducing autophagy

  • Kyeong Eun, Yang (Bio-Chemical Analysis Group, Center for Research Equipment, Korea Basic Science Institute) ;
  • Soo-Bin, Nam (Research Center for Materials Analysis, Korea Basic Science Institute) ;
  • Minsu, Jang (Division of Biological Science and Technology, Yonsei University) ;
  • Junsoo, Park (Division of Biological Science and Technology, Yonsei University) ;
  • Ga-Eun, Lee (BRL & BK21-4th Team, College of Pharmacy, The Catholic University of Korea) ;
  • Yong-Yeon, Cho (BRL & BK21-4th Team, College of Pharmacy, The Catholic University of Korea) ;
  • Byeong-Churl, Jang (Department of Molecular Medicine, College of Medicine, Keimyung University) ;
  • Cheol-Jung, Lee (Research Center for Materials Analysis, Korea Basic Science Institute) ;
  • Jong-Soon, Choi (Research Center for Materials Analysis, Korea Basic Science Institute)
  • Received : 2022.06.09
  • Accepted : 2022.11.03
  • Published : 2023.03.02
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Abstract

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

Keywords

Acknowledgement

This research was supported by Korea Basic Science Institute grants (C280320 and C230110) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2021R1F1A1057701).

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