DOI QR코드

DOI QR Code

Antimicrobial, Antioxidant and Cellular Protective Effects of Houttuynia cordata Extract and Fraction

어성초 추출물 및 분획물의 항균, 항산화 및 세포보호활성

  • Yun, Mid Eum (Department of Fine Chemistry, Cosmetic R&D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology) ;
  • Lee, Ye Seul (Department of Fine Chemistry, Cosmetic R&D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology) ;
  • Lee, Yun Ju (Department of Fine Chemistry, Cosmetic R&D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology) ;
  • Park, Young Min (Department of Fine Chemistry, Cosmetic R&D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology) ;
  • Park, Soo Nam (Department of Fine Chemistry, Cosmetic R&D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology)
  • 윤믿음 (서울과학기술대학교 정밀화학과, 화장품종합기술연구소, 코스메틱 융.복합산업 지원센터) ;
  • 이예슬 (서울과학기술대학교 정밀화학과, 화장품종합기술연구소, 코스메틱 융.복합산업 지원센터) ;
  • 이윤주 (서울과학기술대학교 정밀화학과, 화장품종합기술연구소, 코스메틱 융.복합산업 지원센터) ;
  • 박영민 (서울과학기술대학교 정밀화학과, 화장품종합기술연구소, 코스메틱 융.복합산업 지원센터) ;
  • 박수남 (서울과학기술대학교 정밀화학과, 화장품종합기술연구소, 코스메틱 융.복합산업 지원센터)
  • Received : 2018.03.28
  • Accepted : 2018.05.06
  • Published : 2018.08.10

Abstract

This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

본 연구에서는 어성초(Houttuynia cordata)의 추출물 및 분획물을 제조하고 이들에 대한 생리활성과 성분분석을 하였다. 어성초는 50% 에탄올 추출물 및 에틸아세테이트 분획물을 제조하여 실험에 사용하였다. 항균활성 측정결과, S. aureus, B. subtilis에 대한 에틸아세테이트 분획의 MIC 값은 각각 78, $312{\mu}g/mL$으로 나타나 그람 양성균에 대하여 높은 활성을 보이는 것으로 나타났다. 1,1-Diphenyl-2-picrylhydrazyl (DPPH)에 대한 자유라디칼 소거활성($FSC_{50}$)은 50% 에탄올 추출물($27.15{\mu}g/mL$) 및 에틸아세테이트 분획($12.00{\mu}g/mL$)을 측정한 결과 에틸아세테이트 분획에서 활성이 더 크게 나타났다. Luminol-의존성 화학발광법을 이용한 $Fe^{3+}-EDTA/H_2O_2$계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 총항산화능($OSC_{50}$)은 50% 에탄올 추출물($2.91{\mu}g/mL$), 에틸아세테이트 분획($0.983{\mu}g/mL$) 측정 결과 DPPH에 대한 소거활성과 같은 경향성을 보였다. 어성초 추출물의 HaCaT 세포보호효과를 확인하기 위해 UVB 조사 후 세포 내 ROS 소거활성을 확인한 결과, 어성초 에틸아세테이트 분획은 $1.6{\mu}g/mL$부터 농도 의존적으로 ROS 소거활성을 나타내었으며, 최고 농도인 $12.5{\mu}g/mL$에서 54.3%의 감소율을 보였다. 과산화수소로 유도된 세포보호효과 또한, 어성초 에틸아세테이트 분획은 $0.8{\mu}g/mL$부터 세포생존율을 유의적으로 증가시켜 최대 86.9%까지 증가시켰다. 실험에 사용된 어성초 추출물의 에틸아세테이트 분획을 가지고 TLC와 HPLC를 이용한 성분분석을 수행하였다. 그 결과 quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin 및 afzelin이 있음을 확인하였다. 이상의 결과들로부터, 어성초 추출물 및 분획물은 ROS를 소거하고 항균효과를 나타냄으로써 외부 환경으로부터 세포막을 보호할 수 있는 천연 항산화제 및 천연 방부제로서 화장품 분야에서 응용가능성이 있음을 시사하였다.

Keywords

References

  1. Y. Mizutani, S. Mitsutake, K. Tsuji, A. Kihara, and Y. Igarashi, Ceramide biosynthesis in keratinocyte and its role in skin function, Biochimie, 91(6), 784-790 (2009). https://doi.org/10.1016/j.biochi.2009.04.001
  2. H. J. Kim, J. Y. Bae, H. N. Jang, and S. N. Park, Comparative study on the antimicrobial activity of Glycyrrhiza uralensis and Glycyrrhiza glabra extracts with various countries of origin as natural antiseptics, Korean J. Microbiol. Biotechnol., 41(3), 358-366 (2013). https://doi.org/10.4014/kjmb.1307.07003
  3. Y. H. Yeo and C. H. Park, Cosmetics preservation and moisturizing effect by methanol extracts of Scutellaria baicalensis George and Liriope platyphylla, Korean Soc. Biotechnol. Bioeng. J., 29(5), 372-379 (2014).
  4. C. N. Smith and B. R. Alexander, The relative cytotoxicity of personal care preservative systems in Balb/C 3T3 clone A31 embryonic mouse cells and the effect of selected preservative systems upon the toxicity of a standard rinse-off formulation, Toxicol. in Vitro, 19, 963-969 (2005). https://doi.org/10.1016/j.tiv.2005.06.014
  5. E. Y. Choi, Effect of phenoxyethanol and alkane diol mixture on the antimicrobial activity and antiseptic ability in cosmetics, Korean J. Aesthet. Cosmetol., 13(2), 213-220 (2015).
  6. J. P. Routledge, J. Odum, J. Ashby, and J. P. Sumpter, Some alkyl hydroxy benzoate preservatives (parabens) are estrogenic, Toxicol. Appl. Pharmacol., 153, 12-19 (1998). https://doi.org/10.1006/taap.1998.8544
  7. J. Nishihara, T. Nishikawa, T. Kanayame, F. Dakeyame, K. Saito, M. Imagawa, S.Takatori, Y. Kitagawa, S. Hori, and H. Utasumi, Estrogenic activities of 517 chemicals by yeast two hybrid assay, J. Health Sci., 46(4), 282-298 (2000). https://doi.org/10.1248/jhs.46.282
  8. F. A. Anderson, Final amaended report on the safety assessment of methylparaben, ethylparaben, propylparaben, isopropylparaben, butylparaben, isobutylparaben and benzylparaben as used in cosmetic products, Int. J. Toxicol., 27(4), 1-82 (2008). https://doi.org/10.1177/109158180802704s01
  9. S. N. Park, Skin aging and antioxidants, J. Soc. Cosmet. Sci. Korea, 23(1), 75-132 (1997).
  10. E. H. Witt, P. Motchnik, and L. Packer, Evidence for UV light as an oxidative stressor in skin. In: J. Fuchs and L. Packer (eds.), Oxidative Stress in Dermatology, 29-47, Dekker, NY, USA (1993).
  11. L. Packer, Ultraviolet radiation (UVA, UVB) and skin antioxidants. In: C. A. Rice-Evans and R. H. Burdon (eds.), Free Radical Damage and Its Control 239-255, Elsevier, NY, USA (1994).
  12. H.-U. Simon, A. Haj-Yehia, and F. Levi-Schaffer, Role of reactive oxygen species (ROS) in apoptosis induction, Apoptosis, 5(5), 415-418 (2000). https://doi.org/10.1023/A:1009616228304
  13. V. Afonso, R. Champy, D. Mitrovic, P. I. Collin, and A. Lomri, Reactive oxygen species and superoxide dismutases: Role in joint diseases, Joint Bone Spine, 74(4), 324-329 (2007). https://doi.org/10.1016/j.jbspin.2007.02.002
  14. S. N. Park, Protective effect of isoflavone, genistein from soybean on singlet oxygen induced photohemolysis of human erythrocytes, Korean J. Food Sci. Technol., 35(3), 510-518 (2003).
  15. T. F. Slater, Free radical mechanisms in tissue injury, Biochem. J., 222, 1-15 (1984). https://doi.org/10.1042/bj2220001
  16. S. I. Oh, Effect of melatonin on rat skeletal muscles of oophorectomized rat : oxidative stress and anti of oophorectomized rat : oxidative stress and antioxidative enzyme activities, Korean J. Gerontol., 12, 1-10 (2002).
  17. R. Kohen, Skin antioxidants: their role in aging and in oxidative stress - New approaches for their evaluation, Biomed. Pharmacother., 53(4), 181-192 (1999). https://doi.org/10.1016/S0753-3322(99)80087-0
  18. K. Scharffetter-Kochanek, Photoaging of the connective tissues of skin: Its prevention and therapy, Adv. Pharmacol., 38, 639-655 (1996).
  19. H. Masaki, S. Sakaki, T. Atsumi, and H. Sakurai, Active oxygen scavenging activity of plants extracts, Biol. Pharm. Bull., 18(1), 162-166 (1995). https://doi.org/10.1248/bpb.18.162
  20. D. Bown, Royal Horticultural Society Encyclopedia of Herbs and Their Uses, p. 424, Dorling Kindersley, London, UK (1995).
  21. H. R. Jeong, J. H. Kwak, J. H. Kim, G. N. Choi, C. H. Jeong, and H. J. Heo, Antioxidant and neuronal cell protective effects of an extract of Houttuynia cordata Thunb (a culinary herb), Korean J. Food Preserv., 17(5), 720-726 (2010).
  22. K. Y. Kim, D. O. Chung, and H. J. Chung, Chemical composition and antimicrobial activities of Houttuynia cordata thumb, Korean J. Food Sci. Technol., 29(3), 400-406 (1997).
  23. G. S. Kim, D. H. Kim, J. J. Lim, J. J. Lee, D. Y. Han, W. M. Lee, W. C. Jung, W. G. Min, C. G. Won, M. H. Rhee, H. J. Lee, and S. Kim, Biological and antibacterial activities of the natural herb Houttuynia cordata water extract against the iIntracellular bacterial pathogen Salmonella within the RAW 264.7 macrophage, Biol. Pharm. Bull, 31(11), 2012-2017 (2008). https://doi.org/10.1248/bpb.31.2012
  24. J. H. Song, M. J. Kim, H. D. Kwon, and I. H. Park, Antimicrobial activity of fractional extracts from Houttuynia cordata root, J. Korean Soc. Food Sci. Nutr., 32, 1053-1058 (2003). https://doi.org/10.3746/jkfn.2003.32.7.1053
  25. J. Kim, H. S. Ryu, J. H. Shin, and H. S. Kim, In vitro and ex vivo supplementation of Houttuynia cordata extract and immunomodulating effect in mice, J. Korean Soc. Food Sci. Nutr., 34, 167-175 (2005). https://doi.org/10.3746/jkfn.2005.34.2.167