Journal of the Korean Applied Science and Technology (한국응용과학기술학회지)

The Korean Society of Applied Science and Technology (한국응용과학기술학회)
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Protective Effects of Trifolium pratense L. Extract against H2O2-induced Oxidative Stress in HaCaT Keratinocytes

인간 피부각질세포에서 Hydrogen peroxide로 유도된 산화적 스트레스에 대한 붉은 토끼풀 추출물의 세포 보호 효과

  • Mi Song Shin (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • You Kyeong Lee (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Seo Young Choi (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Ji Sun Hwang (New Drug Development Center, Daegu–Gyeongbuk Medical Innovation Foundation, K-MEDI hub) ;
  • Parkyong Song (Department of Convergence Medicine, Pusan National University School of Medicine) ;
  • Hyeon Cheal Park (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Keun Ki Kim (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Hong-Joo Son (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Yu-Jin Kim (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University) ;
  • Kwang Min Lee (Department of Life Science and Environmental Biochemistry, Life and Industry Convergence Research Institute, Pusan National University)
  • 신미송 (부산대학교 생명환경화학과) ;
  • 이유경 (부산대학교 생명환경화학과) ;
  • 최서영 (부산대학교 생명환경화학과) ;
  • 황지선 (대구경북첨단의료산업진흥재단 신약개발지원센터) ;
  • 송박용 (부산대학교 의과대학 융합의학교실) ;
  • 박현철 (부산대학교 생명환경화학과) ;
  • 김근기 (부산대학교 생명환경화학과) ;
  • 손홍주 (부산대학교 생명환경화학과) ;
  • 김유진 (부산대학교 생명환경화학과) ;
  • 이광민 (부산대학교 생명환경화학과)
  • Received : 2023.03.24
  • Accepted : 2023.04.19
  • Published : 2023.04.30
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Abstract

Oxidative stress plays a significant role in the pathogenesis of various skin conditions, resulting in cellular and tissue damage that can contribute to the development of skin tone unevenness, roughness and wrinkles. In this study, we found that Trifolium pratense L. extract (TE) attenuated oxidative-induced damage in HaCaT cells and elucidated the underlying molecular mechanism. Our finding demonstrated that TE effectively protected HaCaT cells against H2O2-induced cell death by inhibiting caspase-3 activation, downregulating Bax and upregulating Bcl-2, and attenuating the activation of three mitogen-activated protein kinases (MAPKs). Our results suggest that TE has remarkable cytoprotective properties against oxidative damage in HaCaT cells and could serve as a complementary or alternative approach to prevent and treat skin damage.

산화적 스트레스는 세포 및 조직 손상을 통해 피부의 탄력 및 보습 기능 저하, 피부 노화 촉진을 비롯한 다양한 피부질환을 일으킨다. 본 연구의 목적은 인간 피부각질세포 (HaCaT keratinocyte)에서 산화적 스트레스에 대한 붉은 토끼풀 추출물의 효능을 검토하여, 피부에 효과적으로 사용할 수 있는 기능성 소재로서의 활용 여부를 확인하고자 하였다. 본 연구에서는 붉은 토끼풀 추출물이 인간 피부각질세포에서 산화적 스트레스에 따른 세포사를 억제시키는 것을 확인하여, 이를 조절하는 보호기전을 규명하였다. 이는 붉은 토끼풀 추출물이 Caspase-3 비활성, 세포사 촉진단백질 Bax 발현 억제, 세포생존 촉진단백질 Bcl-2 발현 증가 및 MAPK 신호전달계 단백질의 인산화 억제를 통해 H2O2에 의해 유도된 산화적 스트레스를 보호할 수 있다는 것을 확인하였다. 따라서 붉은 토끼풀 추출물은 피부의 산화적 손상을 감소시키는 유용한 소재로 평가되며, 이는 피부보호 및 미용을 위한 다양한 제품 및 산업에 활용 가능성이 높은 것으로 판단된다.

Keywords

Acknowledgement

이 과제는 부산대학교 기본연구지원사업(2년)에 의하여 연구되었음

References

  1. D. Roosterman, T. Goerge, S. W. Schneider, N. W. Bunnett, M. Steinhoff, "Neuronal control of skin function: the skin as a neuroimmunoendocrine organ", Physiol Rev., Vol.86, No.4 pp. 1309-1379, (2006).  https://doi.org/10.1152/physrev.00026.2005
  2. V. S. Sylvie, F. Bonte, "Skin hydration: a review on its molecular mechanisms", J. Cosmetic Dermatol, Vol.6, No.2 pp. 75-82, (2007).  https://doi.org/10.1111/j.1473-2165.2007.00300.x
  3. S. -H. Seo, M. O. Choi, "Protectvie effects of Lonicerae Japonicae Flos against hydrogen peroxidase-induced oxidative stress on Human keratinocyte, HaCaT cells", Kor. J. Herbology, Vol.28, No.4 pp. 57-62, (2013).  https://doi.org/10.6116/kjh.2013.28.4.57
  4. J. -A. Park, "Antioxidant effects of Rumex crispus L. leaf extracts and protective effects on Human HaCaT Keratinocyte", J. kor. Soc.B&A, Vol.12, No.2 pp. 189-198, (2011). 
  5. M. Ott, V. Gogvadze, S. Orrenius, B. Zhivotovsky, "Mitochondria, oxidative stress and cell death", Apoptosis, Vol.12, No.5 pp. 913-922, (2007).  https://doi.org/10.1007/s10495-007-0756-2
  6. S. Parthasarathy, D. Steinberg, J. L. Witztum, "The role of oxidized low-density lipoproteins in the pathogenesis of atherosclerosis", Annual Review of Medicine, Vol.43, No.1 pp. 219-225, (1992).  https://doi.org/10.1146/annurev.me.43.020192.001251
  7. F. R. Laurindo, P. L. da Luz, L. Uint, T. F. Rocha, R. G. Jaeger, E. A. Lopes, "Evidence for superoxide radical-dependent coronary vasospasm after angioplasty in intact dogs", Circulation, Vol.83, No.5 pp. 1705-1715, (1991).  https://doi.org/10.1161/01.CIR.83.5.1705
  8. W. J. Cho, H. S. Yoon, Y. H. Kim, J. M. Kim, I. J. Yoo, M. D. Han, I. S. Bang, "Cytoprotective Effects and Gene Expression Patterns Observed Based on the Antioxidant Activity of Lonicera japonica Extract", Journal of Life Science, Vol.23, No.8 pp. 989-997, (2013).  https://doi.org/10.5352/JLS.2013.23.8.989
  9. B. Dan, G. Andrew, "Chinese herbal medicins. Materia Media.", pp. 85-86, WA : Eastland Press, (1986). 
  10. K. C. Huang, "The Pharmacology of Chinese herbs (2nd Ed.)", pp 8-9, FL : CRC., (1998). 
  11. Y. N. Lee, "New Flora of Korea. Vol. II.", pp. 418-421, Kyohak Publishing Co., Ltd, Seoul., (2006). 
  12. Korea Beekeeping Association, "The Korea Beekeeping Bulletin. Serial No. 320", pp. 26-27, Korea Beekeeping Association, (2007). 
  13. S. Ahmad, A. Zeb, "Phytochemical profile and pharmacological properties of Trifolium repens", Journal of Basic and Clinical Physiology and Pharmacology, Vol.32, No.1 pp. 20200015, (2021). 
  14. L. Krenn, D. H. Paper, "Inhibition of angiogenesis and inflammation by an extract of red clover (Trifolium pratense L.)", Phytomedicine Int. J. Phytother. Phytopharm, Vol.16, No.12 pp. 1083-1088, (2009).  https://doi.org/10.1016/j.phymed.2009.05.017
  15. T. Mosmann, "Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays", J Immunol Methods, Vol.16, No.1-2 pp. 55-63, (1983).  https://doi.org/10.1016/0022-1759(83)90303-4
  16. C. Frippiat, Q. M. Chen, S. Zdanov, J. P. Magalhaes, J. Remacle, O. Toussaint, "Subcytotoxic H2O2 stress triggers a release of transforming growth factor-beta 1, which induces biomarkers of cellular senescence of human diploid fibroblasts", J. Biol. Chem, Vol.276, No.4 pp. 2531-2537, (2001).  https://doi.org/10.1074/jbc.M006809200
  17. K. B. Beckman, B. N. Ames, "The free radical theory of aging natures", Physiological reviews, Vol.78, No.2 pp. 547-581, (1988).  https://doi.org/10.1152/physrev.1998.78.2.547
  18. E. S. Alnemri, D. J. Livingston, D. W. Nicholson, G. Salvesen, N. A. Thornberry, W. W. Wong, J. Yuan, "Human ICE/CED-3 protease nomenclature", Cell, Vol.87, No.2 p. 171, (1996). 
  19. B. S. Jeon, N. G. Kholodilov, T. F. Oo, S. Y. Kim, K. J. Tomaselli, A. Srinivasan, L. Stefanis, R. E. Burke, "Activation of caspase-3 in developmental models of programmed cell death in neurons of the substantia nigra", J Neurochem., Vol.73, No.1 pp. 322-333, (1999).  https://doi.org/10.1046/j.1471-4159.1999.0730322.x
  20. A. Hartmann, S. Hunot, P. P. Michel, M. P. Muriel, S. Vyas, B. A. Faucheux, A. Mouatt-Prigent, H. Turmel, A. Srinivasan, M. Ruberg, G. I. Evan, Y. Agid, E. C. Hirsch, "Caspase-3: A vulnerability factor and final effector in apoptotic death of dopaminergic neurons in Parkinson's disease", Proc Natl Acad Sci U S A., Vol.97, No.6 pp. 2875-2880, (2000).  https://doi.org/10.1073/pnas.040556597
  21. T. Matsura, M. Kai, Y. Fujii, H. Ito, K. Yamada, "Hydrogen peroxide-induced apoptosis in HL-60 cells requires caspase-3 activation", Free Radic Res., Vol.30, No.1 pp. 73-83, (1999).  https://doi.org/10.1080/10715769900300081
  22. G. L. Johnson, R. Lapadat, "Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases", Science (New York, N.Y.), Vol.298, No.5600 pp. 1911-1912, (2002).  https://doi.org/10.1126/science.1072682
  23. H. K. Kim, "Role of ERK/MAPK signalling pathway in antiinflammatory effects of Ecklonia cava in activated human mast cell line cells", Asian Pacific Journal of Tropical Medicine, Vol7, No.9 pp. 703-708, (2014).  https://doi.org/10.1016/S1995-7645(14)60120-6
  24. K. Z. Guyton, Y. Liu, M. Gorospe, Q. Xu, N. J. Holbrook, "Activation of mitogen-activated protein kinase by H2O2 : Role in cell survival following oxidant injury", J Biol Chem., Vol.271, No.8 pp. 4138-4142, (1996).  https://doi.org/10.1074/jbc.271.8.4138
  25. I. Ferrer, R. Blanco, M. Carmona, B. Puig, M. Barrachina, C. Gomez, S. Ambrosio, "Active, phosphorylation-dependent mitogen-activated protein kinase (MAPK/ERK), stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), and p38 kinase expression in Parkinson's disease and Dementia with Lewy bodies", Journal of neural transmission (Vienna, Austria : 1996), Vol.108, No.12 pp. 1383-1396, (2001).  https://doi.org/10.1007/s007020100015
  26. H. Nakano, A. Nakajima, S. Sakon-Komazawa, J. -H. Piao, X. Xue, K. Okumura, "Reactive oxygen species mediate crosstalk between NF-kappaB and JNK", Cell Death Differ, Vol.13, No.5 pp. 730-737, (2006).  https://doi.org/10.1038/sj.cdd.4401830
  27. M. S. Cooke, M. D. Evans, M. Dizdaroglu, J. Lunec, "Oxidative DNA damage: mechanisms, mutation, and disease", FASEB J., Vol.17, No.10 pp. 1195-1214, (2003).  https://doi.org/10.1096/fj.02-0752rev
  28. B. M. Rim, C. W. Rim, J. Y. Choi, Y. S. Chung, H. G. Jeong, "Effects of Lonicera japonica extract as a biological response modifier", Environ Mut Car., Vol.12, No.1 pp. 45-54, (1992). 
  29. Z. H. Luo, "The combined modulating effects of cerium nitrate with certain Chinese traditional drugs on altered cell-mediated immunities in scald mice", Zhonghua Wai Ke Za Zhi., Vol.28, No.9 pp. 562-575, (1990).