DOI QR코드

DOI QR Code

Effect of Fermented Soybean on the Proliferation and Growth in HaCaT and Fibroblast Cell

대두 발효물이 인간 유래 피부세포의 증식 및 성장에 미치는 영향

  • Kim, Eun-Joo (Department of Alternative Medicine Graduate School Kyonggi University) ;
  • Han, Myung-Ryun (Department of Baking Science & Technology Hyejeon University) ;
  • Lee, So-Young (Department of Alternative Medicine Graduate School Kyonggi University) ;
  • Kim, Ae-Jung (Department of Alternative Medicine Graduate School Kyonggi University)
  • 김은주 (경기대학교 일반대학원 대체의학과) ;
  • 한명륜 (혜전대학교 제과제빵과) ;
  • 이소영 (경기대학교 일반대학원 대체의학과) ;
  • 김애정 (경기대학교 일반대학원 대체의학과)
  • Received : 2020.12.18
  • Accepted : 2021.02.05
  • Published : 2021.02.28

Abstract

This study was undertaken to determine the effect of fermented soybean extract and its fractions on skin cell proliferation and growth. The extract was procured by the pepsin and Lactobacillus rhamnosus fermentation of soybean. LC-MS analysis was performed subsequent to soybean fermentation, and cell viability was measured by the WST-1 assay. Cell proliferation was observed to increase after exposing cells to the fermented soybean extract and its fractions at all concentrations tested (0~2,000 ㎍/mL). In particular, compared to the normal control group and 120 % proliferation of the EGF (epidermal growth factor) positive control group, 160~180 % cell proliferation was achieved at 800 ㎍/ml, indicating the excellent potential as an application material for skin aging inhibition and skin cell regeneration. In addition, we also examined the effects of fermented soybean extract and its fractions on wound healing ability, in HaCaT cells and fibroblasts. Our results indicate excellent cell migration abilities after treatment with fermented soybean extract and its fractions, as compared to the control treatment. Similar cell migration abilities were observed in the positive control group (EGF). Taken together, our results indicate that fermented soybean extract and its fractions (F4 and F5) exert amelioratory effects as a natural material for skin.

본 연구는 대두 단백질을 효소(pepsin) 분해 및 미생물(Lactobacillus Rhamnosus) 발효를 통해 생성된 저분자 생리활성 펩티드로 제조하여 인간 유래 피부세포에서 세포활성에 미치는 효과를 확인하고, 천연유래 기능성 미용소재로서의 이용 가능성을 확인하고자 하였다. 연구를 수행하기 위해 대두발효물을 제조하였고, 대두발효물에 대해 LC-MS 분석을 하였으며, WST-1 assay의 방법을 통해 cell viability를 측정하였다. 세포독성을 측정한 결과, 0~2,000 ㎍/mL의 모든 농도 구간에서 세포독성이 나타나지 않았다. 특히 800 ㎍/mL의 농도에서는 대조군에 비해 160~180 %, 양성대조군으로 사용한 EGF(epidermal growth factor)에 비해 120 %에서는 세포증식 효과가 나타나 피부노화 억제 및 피부세포 재생 소재로서의 뛰어난 가능성을 보여주었다. 또한 대두발효물과 그 분획물의 상처 치유 능력을 확인하기 위해 세포이동 실험을 수행한 결과, 대두발효물과 분핵물 중 F4와 F5가 대조군에 비해 우수한 세포이동능을 보여주었고, EGF와는 유사하거나 그 이상의 효과를 보여주었다. 결론적으로 대두발효물과 분핵물(F4 and F5)의 피부개선효과가 유사하게 나타나 피부미용 소재 및 피부 질환 치료의 소재로의 활용이 기대된다.

Keywords

References

  1. D. Agyei, "Bioactive proteins and peptides from soybeans", Recent Patents on Food, Nutrition & Agriculture, Vol.7, No.2, pp.100-107, Agu. 2015. DOI: https://doi.org/10.2174/221279840766615062913414
  2. S. P. Hong, U. H. Choi, S. G. An, "Skin barrier and antimicrobial activity", The Journal of Skin Barrier Research, Vol.11, No.1, pp.55-63, Jun. 2009.
  3. H. Y. Yu, I. J. Yang, V. R Lincha, I. S. Park, D. U. Lee, H. M. Shin, "The Effects of the Fruits of Foeniculum vulgare on Skin Barrier Function and Hyaluronic Acid Production in HaCaT Keratinocytes", Journal of life science, Vol.25, No.8, pp.880-888, Agu. 2015. DOI: http://dx.doi.org/10.5352/JLS.2015.25.8.880
  4. H. S. Jung, M. Y. Song, H. S. Kim, H. H. Seo, J. H. Lee, K. R. Lee, I. Hong, S. H. Moh. "Development of Anti-Wrinkle Materials using Galloyl-Peptide Derivatives", Journal of the Korea Academia-Industria, Vol.16, No.8, pp.5452-5457, Agu. 2015. DOI: http://dx.doi.org/10.5762/KAIS.2015.16.8.5452
  5. M. A. Ruiz, B. Clares, M. E. Morales, V. Gallardo. "Evaluation of the anti-wrinkle efficacy of cosmetic formulations with an anti-aging peptide (Argireline®)", ARS Pharmaceutica, Vol.50, No.4, pp.168-176, Nov. 2009.
  6. B. F. Gibbs, A. Zougman, R. Masse, C. Mulligan. "Produc-tion and characterization of bioactive peptides from soy hydrolysate and soy-fermented food", Food Research International, Vol.37, No.2, pp.123-131, Mar. 2004. DOI: https://doi.org/10.1016/j.foodres.2003.09.010
  7. W. Wang, N. A. Bringe, M. A. Berhow, E. Gonzalez de Mejia. "β-Conglycinins among Sources of Bioactives in Hydrolysates of Different Soybean Varieties That Inhibit Leukemia Cells in Vitro", Journal of Agricultural and Food Chemistry, Vol.56, No.11, pp.4012-4020, Mar. 2008. DOI: https://doi.org/10.1021/jf8002009
  8. E. J. Na, J. S. Moon. "Studies on the Biological Activi-ties of the Bioconversioned Soybean Extracts", Jounal of The Korean Society of cosmetology, Vol.21, No.1, pp.82-92, Feb. 2015.
  9. A. Kannan, N. S. Hettiarachchy, J. O. Lay, R. Liyanage. "Human cancer cell proliferation inhibition by a pen-tapeptide isolated and characterized from rice bran", Peptides, Vol.31, No.9, pp.1629-1634, Sep. 2010. DOI: https://doi.org/10.1016/j.peptides.2010.05.018
  10. B. F. Gibbs, A. Zougman, R. Masse, C. Mulligan. "Production and characterization of bioactive peptides from soy hydrolysate and soy-fermented food", Food Research International, Vol.37, No.2, pp.123-131, Mar. 2004. DOI: https://doi.org/10.1016/j.foodres.2003.09.010
  11. M. Forster, M. A. Bolzinger, H. Fessi, S. Briancon. "Topical delivery of cosmetics and drugs. Molecular aspects of percutaneous absorption and delivery", European Journal of Dermatology, Vol.19, No.4, pp.309-323, Agu. 2009. DOI: https://doi.org/10.1684/ejd.2009.0676
  12. B. Forslin. "A domain mosaic model of the skin barrier", Acta Dermato-venereologica, Vol.74, No.1, pp.1-6, Jan. 1994.
  13. Z. Nemes, P. M. Steinert. "Brick and mortar of the epidermal barrier", Experimental & Molecular Medicine, Vol.31, No.1, pp.5-19, Mar. 1999. DOI: https://doi.org/10.1038/emm.1999.2
  14. S. Ioannis, F. Demosthenes, V. Katerina, G. T. Andreas, K. Valentinos, B. Evangelos. "Cyanobacterial cyclopeptides as lead compounds to novel targeted cancer drugs", Marine Drugs, Vol.8, No.3, pp.629-657, Mar. 2010. DOI: https://doi.org/10.3390/md8030629
  15. J. K. Kim, J. H. Lee, I. H. Bae, D. B. Seo, S. J. Lee. "Beneficial Effect of a Collagen Peptide Supple-ment on the Epidermal Skin Barrier", Korean Journal of Food Science and Technology, Vol.43, No.4, pp.458-463, Agu. 2011. DOI: https://doi.org/10.9721/KJFST.2011.43.4.458
  16. S. Sakai, T. Sayo, S. Kodama, S. N. Inoue. "N-Methyl-L-serine stimulates hyaluronan production in human skin fibroblasts", Skin Pharmacol. Appl", Skin Phar-macology and Physiology, Vol.12, No.5, pp.276-283, Oct. 1999. DOI: https://doi.org/10.1159/000066253
  17. J. C. Lee, E. Lee, Y. C. Lee, H. Oh, H. S. Yoon, T. K. Ha, E. H. Hong. "Effects of Fructus foeniculi extract on recovering liver function", The Korea Journal of Herbology, Vol.22, No.4, pp.213-218, Dec. 2007.
  18. M. C. Kwon, C. H. Kim, H. S. Kim, A. Q. Syed, B. Y. Hwang, H. Y. Lee. "Anti-wrinkle Activity of Low Molecular Weight Peptides Derived from the Collagen Isolated from Asterias amurensis", Korean Journal of Food Science and Technology, Vol.39, No.6, pp.625-629, Oct. 2007.
  19. S. J. Lee, H. R. Choi, J. C. Lee, H. J. Park, H. K. Lee, J. T. Jeong, T. B. Lee. "The Anti-aging Effects of Various Berries in the Human Skin Keratinocyte (HaCaT) Cells", Korean Journal of Food Science and Technology, Vol.46, No.2, pp.198-204, Apr. 2014. DOI: https://doi.org/10.9721/KJFST.2014.46.2.198
  20. A. V. Rawlings, C. R. Harding. "Moisturization and skin barrier function", Dermatologic Therapy, Vol.17, No.1, pp.43-48, Jan. 2004. DOI: https://doi.org/10.1111/j.1396-0296.2004.04S1005.x
  21. F. J. Smith, A. D. Irvine, A. Terron-Kwiatkowski, A. Sandilands, L. E. Campbell, Y. Zhao, H. Liao, A. T. Evans, D. R. Goudie, S. Lewis-Jones, G. Arseculeratne, C. S. Munro, A. Sergeant, G. O'Regan, S. J. Bale, J. G. Compton, J. J. DiGiovanna, R. B. Presland, P. Fleckman, W. H. McLean. "Loss-of-function mutations in the gene encoding filaggrin cause ichthyosis vulgaris", Nature Genetics, Vol.38, No.3, pp.337-342, Jan. 2006. DOI: https://doi.org/10.1038/ng1743
  22. R. L. Gallo, M. Murakami, T. Ohtake, M. Zaiou. "Biology and clinical relevance of naturally occurring antimicro-bial peptides", Journal of Allergy and Clinical Immu-nology, Vol.110, No.6, pp.823-831, Sep. 2002. DOI: https://doi.org/10.1067/mai.2002.129801
  23. J. S. Kim, E. J. Bak, B. C. Lee, Y. S. Kim, J. B. Park, I. G. Choi. "Neuregulin induces HaCaT keratinocyte migration via Racl-mediated NADPH-oxidase activation", Journal of Cellular Physiology, Vol.226, No.11, pp.3014-3021, Feb. 2011. DOI: https://doi.org/10.1002/jcp.22649
  24. G. M. Marcela, R. G. Eva, R. M. Del Carmen, M. E. Rosalva. "Evaluation of the Antioxidant and Anti-proliferative Effects of Three Peptide Fractions of Germinated Soybeans on Breast and Cervical Cancer Cell Lines", Plant Foods for Human Nutrition, Vol.71, No.4, pp.368-374, Jul. 2016. DOI: https://doi.org/10.1007/s11130-016-0568-z
  25. Y. Nzengue, R. Steiman, C. Garrel, "Oxidative stress and DNA damage induced by cadmium in the human keratinocyte HaCaT cell line: role of glutathione in the resistance to cadmium", Toxicology, Vol.243, pp.193-206, Jan. 2008. DOI: https://doi.org/10.1016/j.tox.2007.10.005
  26. J. Lopez-Garcia, M. Lehocky, P. Humpolicek, P. Saha, "HaCaT keratinocytes response on antimicrobial a telocollagen substrates: extent of cytotoxicity, cell viability and proliferation", J Funct Biomater, Vol.5, No.2, pp.43-57, Apr. 2014. DOI: https://doi.org/10.3390/jfb5020043
  27. J. Jonkman, J. Cathcart, F. Xu, M. Bartolini, J. Amon, K. Stevens, P. Colarusso, "An introduction to the wound healing assay using live-cell microscopy", Cell Adhes. Migr, Vol.8, pp. 440-451, Apr. 2014 https://doi.org/10.4161/cam.36224
  28. H. Y. Yu, I. J. Yang, V. R. Lincha, I. S. Park, D. U. Lee, H. M. Shin. "The Effects of the Fruits of Foe-niculum vulgare on Skin Barrier Function and Hya-luronic Acid Production in HaCaT Keratinocytes", Journal of Life Science, Vol.25, No.8, pp.880-888, Agu. 2015. DOI: http://dx.doi.org/10.5352/JLS.2015.25.8.880
  29. B. McKnight, R. Seidel, R. Moy. "Topical Human Epidermal Growth Factor in the Treatment of Senile Purpura and the Prevention of Dermatoporosis", Journal of Drugs in Dermatology, Vol.14, No.10, pp.1147-1150, Oct. 2015.
  30. K. Kaufmann, G. Thiel. "Epidermal growth factor and thrombin induced proliferation of immortalized human keratinocytes is coupled to the synthesis of Egr-1, a zinc finger transcriptional regulator", Journal of Cellular Biochemistry, Vol.85, No.2, pp.381-391, Mar. 2002. DOI: https://doi.org/10.1002/jcb.10145