Journal of Korean Medicine Rehabilitation (한방재활의학과학회지)

The society of Korean Medicine Rehabilitation (한방재활의학과학회)
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Cellular Aging Inhibitory Effect of Perilla Leaf Extract on D-Galactose Induced C2C12 Myoblasts

D-갈락토스 유도 C2C12 근원세포에 대한 자소엽 추출물의 세포 노화 억제 효과

  • Song-Mi Park (Department of Korean Medicine Rehabilitation, Dong-Eui University Korean Medicine Hospital) ;
  • Sung-Woo Cho (Department of Korean Medicine Rehabilitation, Dong-Eui University Korean Medicine Hospital) ;
  • Yung-Hyun Choi (Research Institute of Korean Medicine, Dong-Eui University)
  • 박송미 (동의대학교 부속 한방병원 한방재활의학과) ;
  • 조성우 (동의대학교 부속 한방병원 한방재활의학과) ;
  • 최영현 (동의대학교 한의학연구소)
  • Received : 2024.03.23
  • Accepted : 2024.04.04
  • Published : 2024.04.30
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Abstract

Objectives We used the D-galactose (D-gal) induced C2C12 myoblast senescence model to investigate whether ethanol extract of Perilla. fructescens leaves (EEPF) could delay cellular senescence and regulate related mechanisms. Methods C2C12 myogenic cells were cultured in an incubator under 37 ℃ and 5% CO2 conditions. EEPF, dried perilla leaves were pulverized and extracted at 1:10 (v/v) at 50 ℃ for 4 hours. Cell counting kit-8 and western blot analysis was performed. Annexin V-FITC apoptosis detection kit and DAPI staining was applied. Catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde analysis kits were used. To measure the level of reactive oxygen species generation, staining and flow cytometry was used. To analyze the mitochondrial activity, membrane potential changes were measured using JC-1. 𝛽-gal activity was analyzed using SA-𝛽-gal staining solution, and DNA damage was analyzed by using 𝛾-H2AX. Quantikine ELISA kit was used to analyze inflammatory cytokine production. Results According to the results of this study, EEPF significantly alleviated the decrease in cell viability in C2C12 cells treated with D-gal and suppressed the decrease in the expression of proliferating cell nuclear antigen. EEPF also markedly blocked D-gal-induced C2C12 cell apoptosis and restored reduced activity of CAT, GSH-Px, T-AOC, SOD. In addition, EEPF suppressed the decrease in 𝛽-galactosidase activity, the induction of DNA damage and the increase in expression of senescence-associated secretory phenotype proteins such as p16, p53 and p21 in D-gal-treated C2C12 cells. Furthermore, EEPF significantly attenuated D-gal-induced production and expression of inflammatory cytokines such as interleukin (IL)-6 and IL-18. Conclusions The results of this study indicate that EEPF can be used as a potential candidate for the prevention and treatment of muscle aging.

Keywords

References

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