Action mechanism of anti-wrinkle effect of Rhamnus yoshinoi methanol extract in human dermal fibroblast and keratinocyte cell lines

  • Hyun Ok Kim
  • Kyoung Ran Shin
  • Byeong-Churl Jang
  • Young Chul KimEmail author
Original Article


Rhamnus yoshinoi is a deciduous broad-leaf bush and endemic species widely found in Korea. Recently, we reported that R. yoshinoi methanol extract (RYME) had excellent antioxidant activity and inhibition of collagenase and elastase activity in cell-free system. In this study, we investigated the ability of RYME to control the mRNA and protein expression levels of the known skin wrinkle-related factors in cultured human dermal fibroblast and keratinocyte cell lines. Treatment with 100 or 200 μg/mL RYME strongly blocked the UVB-induced downregulation of type 1 collagen mRNA expression (p < 0.001) and partially blocked the UVB-induced upregulation of MMP-3 mRNA expression in HaCaT human keratinocytes (p < 0.05 or p < 0.001). Treatment with RYME at 100 μg/mL considerably decreased MMP-1 mRNA expression in UVB-exposed HaCaT cells (p < 0.01). In HaCaT cells, RYME exhibited the potential to improve UV light-induced skin wrinkles. Moreover, RYME selectively inhibited the UVB-induced ERK-1/2 protein phosphorylation in CCD-986sk human dermal fibroblasts at 80 and 160 μg/mL. UV-induced ERK-1/2 protein phosphorylation is one of the major mechanisms of the generation of UV-induced skin wrinkles. Therefore, it is likely that the anti-skin wrinkling effect of RYME could be attributable to selective inhibition of UV induced ERK-1/2 protein phosphorylation.


Anti-wrinkle effect CCD-986sk cells ERK-1/2 protein phosphorylation HaCaT cells MMP-3 Rhamnus yoshinoi 


Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to disclose.


  1. 1.
    Brohem CA, Cardeal LB, Tiago M, Soengas MS, Barros SB, Maria-Engler SS (2011) Artificial skin in perspective: concepts and applications. Pigment Cell Melanoma Res 24:35–50CrossRefGoogle Scholar
  2. 2.
    Martin P (1997) Wound healing—aiming for perfect skin regeneration. Science 276:75–81CrossRefGoogle Scholar
  3. 3.
    Majeed M, Bhat B, Anand S, Sivakumar A, Paliwal P, Geetha KG (2011) Inhibition of UV-induced ROS and collagen damage by Phyllanthus emblica extract in normal human dermal fibroblasts. J Cosmet Sci 62:49–56PubMedGoogle Scholar
  4. 4.
    Quan T, He T, Kang S, Voorhees JJ, Fisher GJ (2004) Solar ultraviolet irradiation reduces collagen in photoaged human skin by blocking transforming growth factor-β type II receptor/smad signaling. Am J Pathol 165:741–751CrossRefGoogle Scholar
  5. 5.
    Nelson AR, Fingleton B, Rothenberg ML, Matrisian LM (2000) Matrix metalloproteinases: biologic activity and clinical impli-cations. J Clin Oncol 18:1135–1149CrossRefGoogle Scholar
  6. 6.
    Park CH, Chung JH (2011) Epidermal growth factor-induced matrix metalloproteinase-1 expression is negatively regulated by p38 MAPK in human skin fibroblasts. J Dermatol Sci 64:134–141CrossRefGoogle Scholar
  7. 7.
    Overall CM (2001) Matrix metalloproteinase substrate binding domains, modules and exosites: overview and experimental strategies. Methods Mol Biol 151:79–120PubMedGoogle Scholar
  8. 8.
    Xu Y, Fisher GJ (2005) Ultraviolet (UV) light irradiation induced signal transduction in skin photoaging. J Dermatol Sci 1:51–58Google Scholar
  9. 9.
    Karin M, Hunter T (1995) Transcriptional control by protein phosphorylation: signal transmission form the cell surface nucleus. Curr Biol 5:747–757CrossRefGoogle Scholar
  10. 10.
    Angel P, Szabowski A, Schorpp-Kistner M (2001) Function and regulation of AP-1 subunits in skin physiology and pathology. Oncogene 20:2413–2423CrossRefGoogle Scholar
  11. 11.
    Jung HY, Shin JC, Park SM, Kim NR, Kwak W, Choi BH (2014) Pinus densiflora extract protects human skin fibroblasts against UVB-induced photoaging by inhibiting the expression of MMPs and increasing type I procollagen expression. Toxicol Rep 1:658–666CrossRefGoogle Scholar
  12. 12.
    Kim HO, Oh CJ, Jang B, Kim YC (2018) Antioxidant activity and anti-wrinkle effect of Rhamnus yoshinoi methanol extract. J Invest Cosmetol 14:421–427Google Scholar
  13. 13.
    Bosch R, Philips N, Suárez-Pérez JA, Juarranz A, Devmurari A, Chalensouk-Khaosaat J, González S (2015) Mechanisms of photoaging and cutaneous photocarcinogenesis, and photoprotective strategies with phytochemicals. Antioxidants 4:248–268CrossRefGoogle Scholar
  14. 14.
    Fisher GJ, Datta SC, Talwar HS, Wang ZQ, Varani J, Kang S, Voorhees JJ (1996) Molecular basis of sun-induced premature skin ageing and retinoid antagonism. Nature 379:335–339CrossRefGoogle Scholar
  15. 15.
    Brenneisen P, Wenk J, Klotz LO, Wlaschek M, Briviba K, Krieg T, Sies H, Scharffetter-Kochanek K (1998) Central role of ferrous/ferric iron in the ultraviolet B irradiation-mediated signaling pathway leading to increased interstitial collagenase (matrix-degrading metalloprotease (MMP)-1) and stromelysin-1 (MMP-3) mRNA levels in cultured human dermal fibroblasts. J Biol Chem 273:5279–5287CrossRefGoogle Scholar
  16. 16.
    Quan T, Qin Z, Xia W, Shao Y, Voorhees JJ, Fisher GJ (2009) Matrix-degrading metalloproteinases in photoaging. J Investig Dermatol Symp Proc 14:20–24CrossRefGoogle Scholar
  17. 17.
    Feng XX, Yu XT, Li WJ, Kong SZ, Liu YH, Zhang X, Xian YF, Zhang XJ, Su ZR, Lin ZX (2014) Effects of topical application of patchouli alcohol on the UV-induced skin photoaging in mice. Eur J Pharm Sci 63:113–123CrossRefGoogle Scholar
  18. 18.
    Scharffetter-Kochanek K, Brenneisen P, Wenk J, Herrmann G, Ma W, Kuhr L, Meewes C, Wlaschek M (2000) Photoaging of the skin from phenotype to mechanisms. Exp Gerontol 35:307–316CrossRefGoogle Scholar
  19. 19.
    Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE (1988) Normal keratinization in a spontaneously immortalized aneuploidy human keratinocyte cell line. J Cell Biol 106:761–771CrossRefGoogle Scholar
  20. 20.
    Blanpain C, Lowry WE, Geoghegan A, Polak L, Fuchs E (2004) Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. Cell 118:635–648CrossRefGoogle Scholar
  21. 21.
    Moon HJ, Park KS, Ku MJ, Lee MS, Jeong SH, Imbs TI, Zvyagintseva TN, Ermakova SP, Lee YH (2009) Effect of Costaria costata fucoidan on expression of matrix metalloproteinase-1 promoter, mRNA, and protein. J Nat Prod 72:1731–1734CrossRefGoogle Scholar
  22. 22.
    Kim MS, Oh GH, Kim MJ, Hwang JK (2013) Fucosterol inhibits matrix metalloproteinase expression and promotes type I procollagen production in UVB-induced HaCaT cells. Photochem Photobiol 89:911–918CrossRefGoogle Scholar
  23. 23.
    Sudel KM, Venzke K, Mielke H, Breitenbach U, Mundt C, Jaspers S, Koop U, Sauermann K, Knußmann-Hartig E, Moll L, Gercken G, Young AR, Stab F, Wenck H, Gallinat S (2005) Novel aspects of intrinsic and extrinsic aging of human skin: beneficial effects of soy extract. Photochem Photobiol Sci 81:581–587CrossRefGoogle Scholar
  24. 24.
    Inomata S, Matsunaga Y, Amano S, Takada K, Kobayashi K, Tsunenaga M, Nishiyama T, Kohno Y, Fukuda M (2003) Possible involvement of gelatinases in basement membrane damage and wrinkle formation in chronically ultraviolet B-exposed hairless mouse. J Invest Dermatol 120:128–134CrossRefGoogle Scholar
  25. 25.
    Robinson MJ, Cobb MH (1997) Mitogen-activated protein kinase pathways. Curr Opin Cell Biol 9:180–186CrossRefGoogle Scholar
  26. 26.
    Rosette C, Karin M (1996) Ultraviolet light and osmotic stress: activation of the JNK cascade through multiple growth factor and cytokine receptors. Science 274:1194–1197CrossRefGoogle Scholar
  27. 27.
    Gum R, Wang H, Lengyel E, Juarez J, Boyd D (1997) Regulation of 92 kDa type IV collagenase expression of the jun aminoterminal kinase- and the extracellular signal-regulated kinase-dependent signaling cascades. Oncogene 14:1481–1493CrossRefGoogle Scholar
  28. 28.
    Chen A, Davis BH (1999) UV irradiation activates JNK and increases αI(I) collagen gene expression in rat hepatic stellate cells. J Biol Chem 274:158–164CrossRefGoogle Scholar
  29. 29.
    Xia Z, Dicknens M, Raingeand J, Davs RJ, Greenberg ME (1995) Opposing effects of ERK and JNK-p38 MAP kinase on apoptosis. Science 270:1326–1331CrossRefGoogle Scholar
  30. 30.
    Ham J, Babij C, Whitfield J, Pfarr CM, Lallemand D, Yaniv M, Rubin LL (1995) A c-Jun dominant negative mutant protects sympathetic neurons against programmed cell death. Neuron 14:927–939CrossRefGoogle Scholar
  31. 31.
    Rittie L, Fisher GJ (2002) UV-light-induced signal cascades and skin aging. Ageing Res Rev 1:705–720CrossRefGoogle Scholar

Copyright information

© Korean Society of Toxicology 2019

Authors and Affiliations

  • Hyun Ok Kim
    • 1
  • Kyoung Ran Shin
    • 2
  • Byeong-Churl Jang
    • 2
  • Young Chul Kim
    • 1
    Email author
  1. 1.Department of Public Health, Graduate SchoolKeimyung UniversityDaeguKorea
  2. 2.Department of Molecular Medicine, College of MedicineKeimyung UniversityDaeguKorea

Personalised recommendations