Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of Saponin with Antioxidant Activity on Matrix Metalloproteinase in Human Dermal Fibroblasts

항산화 효능을 가진 사포닌이 사람섬유아세포에서 기질 금속 단백질 분해효소에 미치는 영향

  • Received : 2011.07.04
  • Accepted : 2011.08.09
  • Published : 2011.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

사포닌은 동양의 전통적인 약재로 널리 알려진 인삼의 주요한 성분이다. 사포닌의 다양한 생물학적 효능이 밝혀져 있으나, 피부재생과 관련된 효능은 현재까지도 명백하지 않다. 본 연구에서는 세포외 시스템에서 사포닌의 항산화 효과 뿐만 아니라 사람 진피 섬유아세포에서 기질금속단백질 분해효소(MMP)에 대한 효능이 조사되었다. 먼저 MTT assay를 이용한 세포생존력에 대한 사포닌의 효능을 조사한 결과, 10 ${\mu}g$/ml 이하의 농도에서 사포닌은 세포생존력을 증가시켰으나 25 ${\mu}g$/ml 이상의 농도에서는 세포독성을 나타내었다. 항산화에 대한 사포닌 효능을 조사한 결과, 1 ${\mu}g$/ml 이상의 농도에서 사포닌은 환원력 뿐만 $H_2O_2$에 대한 억제 효능을 보여주었다. 특히, DNA 산화에 대한 보호 효과도 나타내었다. 더욱이 gelatin 및 casein zymography 시험결과, 사포닌은 MMP-2를 활성화 시키고 MMP-1의 활성을 증가시키는 것으로 보아, 항노화 및 피부재생 약효제로 잠재적인 가능성이 기대된다.

Keywords

References

  1. Ali, M. B., K. W. Yu, E. J. Hahn, and K. Y. Paek. 2005. Differential responses of anti-oxidants enzymes, lipoxygenase activity, ascorbate content and the production of saponins in tissue cultured root of mountain Panax ginseng CA Mayer and Panax quinquefolium L. in bioreactor subjected to methyl jasmonate stress. Plant Sci. 169, 83-92. https://doi.org/10.1016/j.plantsci.2005.02.027
  2. Alscher, R., N. Erturk, and L. Heath. 2002. Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J. Exp. Bot. 53, 1331-1341. https://doi.org/10.1093/jexbot/53.372.1331
  3. Bandyopadhyay, U., D. Das, and R. Banerjee. 1999. Reactive oxygen species: Oxidative damage and pathogenesis. Curr Sci. 77, 658-666.
  4. Beckman, K. B. and B. N. Ames. 1998. The free radical theory of aging matures. Physiol. Rev. 78, 547-581.
  5. Bogenrieder, T. and M. Herlyn. 2003. Axis of evil: molecular mechanisms of cancer metastasis. Oncogene 22, 6524-6536. https://doi.org/10.1038/sj.onc.1206757
  6. Choi, C. W., S. C. Kim, S. S. Hwang, B. K. Choi, H. J. Ahn, M. Y. Lee, S. H. Park, and S. K. Kim. 2002. Antioxidant activity and free radical scavenging capacity between Korean medicinal plants and flavonoids by assay-guided comparison. Plant Sci. 163, 1161-1168. https://doi.org/10.1016/S0168-9452(02)00332-1
  7. Cho, S. J., H. J. Baik., S. S. Lee, I. M. Chung, J. H. Ha, J. S. Kang, H. C Koh,I. C Shin, and C. H. Lee. 2000. Studies on the ROS (Reactive Oxygen Species)-scavenging activities of DDMP saponin isolated from glycine mac (L.) Merrill. J. Appl. Pharmacol. 8, 32-37.
  8. Choi, S. 2002. Epidermis proliferative effect of the Panax ginseng ginsenoside Rb 2. Arch. Pharm. Res. 25, 71-76. https://doi.org/10.1007/BF02975265
  9. Fath, A., P. Bethke, and R. Jones. 2001. Enzymes that scavenge reactive oxygen species are down-regulated prior to gibberellic acid-induced programmed cell death in barley aleurone. Plant Physiol. 126, 156-166. https://doi.org/10.1104/pp.126.1.156
  10. Fukuda, N., H. Tanaka, and Y. Shoyama. 2000. Isolation of the pharmacologically active saponin ginsenoside Rb1 from ginseng by immunoaffinity column chromatography. J. Nat. Prod. 63, 283-285. https://doi.org/10.1021/np990356s
  11. Hansen, M. B., S. E. Nielsen, and K. Berg. 1989. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J. Immunol. Methods 119, 203-210. https://doi.org/10.1016/0022-1759(89)90397-9
  12. Hofmann, U. B., J. R. Westphal, G. N. P. van Muijen, and D. J. Ruiter. 2000. Matrix metalloproteinases in human melanoma. J. Invest. Dermatol. 115, 337-344. https://doi.org/10.1046/j.1523-1747.2000.00068.x
  13. Hollborn, M., C. Stathopoulos, A. Steffen, P. Wiedemann, L. Kohen, and A. Bringmann. 2007. Positive feedback regulation between MMP-9 and VEGF in human RPE cells. Invest. Ophth. Vis. Sci. 48, 4360-4367. https://doi.org/10.1167/iovs.06-1234
  14. Imai, J., N. Ide, S. Nagae, T. Moriguchi, H. Matsuura, and Y. Itakura. 1994. Antioxidant and Radical Scavanging Effects of Aged Garlic Extract and its Constituents. Planta Med. 60, 417-417. https://doi.org/10.1055/s-2006-959522
  15. Kang, J. H., I. H. Han, M. K. Sung, H. Yoo, Y. G. Kim, J. S. Kim, T. Kawada, and R. Yu. 2008. Soybean saponin inhibits tumor cell metastasis by modulating expressions of MMP-2, MMP-9 and TIMP-2. Cancer Lett. 261, 84-92. https://doi.org/10.1016/j.canlet.2007.11.006
  16. Kang, J., M. Sung, T. Kawada, H. Yoo, Y. Kim, J. Kim, and R. Yu. 2005. Soybean saponins suppress the release of proinflammatory mediators by LPS-stimulated peritoneal macrophages. Cancer Lett. 230, 219-227. https://doi.org/10.1016/j.canlet.2004.12.041
  17. Kang, S., S. Cho, J. Chung, C. Hammerberg, G. Fisher, and J. Voorhees. 2005. Inflammation and extracellular matrix degradation mediated by activated transcription factors nuclear factor-{kappa} B and activator protein-1 in inflammatory acne lesions in vivo. Am. J. Pathol. 166, 1691-1699. https://doi.org/10.1016/S0002-9440(10)62479-0
  18. Kim, H. G., I. S. Oh, S. S. So, and C. W. Park. 2001. Effect of VEGF on the secretion of MMP-2 and plasmin from human Keratinocytes cells. Korean J. Biotechnol. Bioeng. 16, 237-240.
  19. Lee, K., S. Hwang, J. Choi, and H. Jeong. 2008. Saponins derived from the roots of Platycodon grandiflorum inhibit HT-1080 cell invasion and MMPs activities: Regulation of NF-[kappa] B activation via ROS signal pathway. Cancer Lett. 268, 233-243. https://doi.org/10.1016/j.canlet.2008.03.058
  20. Maniatis, T. 1989. Molecular cloning: a laboratory manual/J. Sambrook, EF Fritsch, T. Maniatis: New York: Cold Spring Harbor Laboratory Press.
  21. Milne, L., P. Nicotera, S. Orrenius, and M. Burkitt. 1993. Effects of glutathione and chelating agents on copper-mediated DNA oxidation: pro-oxidant and antioxidant properties of glutathione. Arch. Biochem. Biophys. 304, 102-109. https://doi.org/10.1006/abbi.1993.1327
  22. Neufeld, G., T. Cohen, S. Gengrinovitch, and Z. Poltorak. 1999. Vascular endothelial growth factor (VEGF) and its receptors. FASEB J. 13, 9-22.
  23. Oyaizu, M. 1986. Studies on products of the browning reaction. Antioxidative activities of browning reaction products prepared from glucosamine. Jpn. J. Nutr. 44, 307-315. https://doi.org/10.5264/eiyogakuzashi.44.307
  24. Simon, H., A. Haj-Yehia, and F. Levi-Schaffer. 2000. Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis 5, 415-418. https://doi.org/10.1023/A:1009616228304
  25. Sounni, N., M. Janssen, J. Foidart, and A. Noel. 2003. Membrane type-1 matrix metalloproteinase and TIMP-2 in tumor angiogenesis. Matrix Biol. 22, 55-61. https://doi.org/10.1016/S0945-053X(03)00003-9
  26. Stadtman, E. and C. Oliver. 1991. Metal-catalyzed oxidation of proteins. Physiological consequences. J. Biol. Chem. 266, 2005-2008.
  27. Vihinen, P. and V. Kahari. 2002. Matrix metalloproteinases in cancer: prognostic markers and therapeutic targets. Int. J.Cancer 99, 157-166. https://doi.org/10.1002/ijc.10329
  28. Wang, C., D. Shi, and H. Yin. 2007. Effect of panax quinquefolius saponin on angiogenesis and expressions of VEGF and bFGF in myocardium of rats with acute myocardial infarction]. Zhongguo Zhong Yi Yan Jiu Yuan Zhu Ban. 27, 331-334.
  29. Yoshiki, Y. and K. Okubo. 1995. Active oxygen scavenging activity of DDMP (2, 3-dihydro-2, 5-dihydroxy-6-methyl-4H-pyran-4-one) saponin in soybean seed. Biosci. Biotechnol. Biochem. 59, 1556-1557. https://doi.org/10.1271/bbb.59.1556
  30. Zhao, H. and H. Yang. 2008. Exogenous polyamines alleviate the lipid peroxidation induced by cadmium chloride stress in Malus hupehensis Rehd. Sci. Hortic. 116, 442-447. https://doi.org/10.1016/j.scienta.2008.02.017

Cited by

  1. The Effects on Anti-inflammatory Action in HaCaT Cells and Inhibiting Sebum Secretion in SEB-1 Cells by Gleditsiae Fructus Extract vol.30, pp.2, 2016, https://doi.org/10.7778/jpkm.2016.30.2.096
  2. A New Triterpenoid Saponin from the Tropical Marine Sponge Lipastrotethya sp. vol.38, pp.4, 2016, https://doi.org/10.4217/OPR.2016.38.4.287