Screening of Crude Drugs for the Inhibitory Effect on $NF-{\kappa}B$ Activation in Transfectant HaCaT Cells

형질전환된 각질형성세포에서 생약추출물에 의한 $NF-{\kappa}B$ 활성화 억제효과 탐색

  • Ahn, Kwang-Seok (Natural Products Research Institute, College of Pharmacy, Seoul National University) ;
  • Kim, Seong-Kie (Korea Research Institute of Chemical Technology) ;
  • Moon, Ki-Young (Department of Clinical Pathology, Kwangju Health College) ;
  • Hahn, Bum-Soo (Metabolic Engineering Division, National Institute of Agricultural Biotechnology) ;
  • Kang, Sam-Sik (Natural Products Research Institute, College of Pharmacy, Seoul National University) ;
  • Kim, Yeong-Shik (Natural Products Research Institute, College of Pharmacy, Seoul National University)
  • 안광석 (서울대학교 천연물과학연구소/약학대학) ;
  • 김성기 (한국화학연구원) ;
  • 문기영 (광주보건대학 임상병리과) ;
  • 한범수 (농업생명공학연구원 신기능소재개발과) ;
  • 강삼식 (서울대학교 천연물과학연구소/약학대학) ;
  • 김영식 (서울대학교 천연물과학연구소/약학대학)
  • Published : 2003.06.30

Abstract

$NF-{\kappa}B$ (nuclear factor-kappa B) plays a particularly central role in epidermal biology. It has been established that ultraviolet radiation (UVR) is one of the mechanisms to induce the activation of $NF-{\kappa}B$ in human skin. We previously demonstrated that melanogenic inhibitors may act through the inhibition of $NF-{\kappa}B$ activation in keratinocytes. In order to find another type of melanogenic inhibitors of $NF-{\kappa}B$ activation, various kinds of the extracts from crude drugs $(30\;{\mu}g/ml)$ were preincubated with transfectant HaCaT cells for 3 hrs and then UVR $(60\;mj/cm^2)$ was irradiated. UVR-exposed cells were incubated for another 6 hrs to measure the $NF-{\kappa}B$ activity. $NF-{\kappa}B$ activation was measured with the secreatory alkaline phosphates (SEAP) reporter gene assay using a fluorescence detection method. Among natural products, Lycium chinense, Acanthopanax senticosus, Angelica koreana, Kalopanax pictus and Asparagus cochinchinensis were the most potent inhibitors of $NF-{\kappa}B$ activation by UVR. These observations suggest that some crude drugs might act partially through the modulation of the synthesis of melanotrophic factors to decrease melanogenesis in keratinocytes.

Keywords

References

  1. Aberdam, E., Romero, C. and Ortonne, J.P (1993). Repeated UVB irradiations do not have the same potential to promote stimulation of melanogenesis in cultured normal human melanocytes. J. Cell. Sci. 106(Pt 4), 1015-1022
  2. Ramirez-Bosca, A, Bemd, A, Wemer, R, Dold, K and Holzmann, H. (1992). Effect of the dose of ultraviolet radiation on the pigment formation by human melanocytes in vitro. Arch. Dermatol. Res. 284(6): 358-362 https://doi.org/10.1007/BF00372040
  3. Seiberg, M., Paine, C., Sharlow, E., Andrade-Gordon, P, Costanzo, M., Eisinger, M. and Shapiro, S.S. (2000). The protease-activated receptor 2 regulates pigmentation via keratinocyte-melanocyte interactions. Exp. Cell. Res. 254(1): 25-32 https://doi.org/10.1006/excr.1999.4692
  4. Donatien, P, Surleve-Bazeille, J.E., Thody, A .J. and Taieb, A (1993). Growth and differentiation of normal human melanocytes in a TPA-free, cholera toxin-free, low-serum medium and influence of keratinocytes. Arch. Dermatol. Res. 285(7) 385-392 https://doi.org/10.1007/BF00372130
  5. Saliou, C., Kitazawa, M., McLaughlin, L., Yang, J.P, Lodge, J.K, Tetsuka, T., Iwasaki, K, Cillard, J., Okamoto, T. and Packer, L. (1999). Antioxidants modulate acute solar ultraviolet radiation-induced NF-kappa-B activation in a human keratinocyte cell line. Free. Radic. Biol. Med. 26(1-2): 174-183 https://doi.org/10.1016/S0891-5849(98)00212-3
  6. Baeuerle, P.A and Henkel, T. (1994). Function and activation of NF-kappa B in the immune system. Annu. Rev. Immunol. 12: 141-179 https://doi.org/10.1146/annurev.iy.12.040194.001041
  7. Baeuerle, P.A and Baichwal, V .R (1997). NF-kappa B as a frequent target for immunosuppressive and anti-inflammatory molecules. Adv. Immunol. 65: 111-137 https://doi.org/10.1016/S0065-2776(08)60742-7
  8. Wulczyn, F.G., Krappmann, D. and Scheidereit, C. (1996). The NF-kappa B/Rel and I kappa B gene families: mediators of immune response and inflammation. J. Mol. Med. 74(12): 749-769 https://doi.org/10.1007/s001090050078
  9. Auphan, N., DiDonato, J.A, Rosette, C, Helmberg, A and Karin, M. (1995). Immunosuppression by glucocorticoids: inhibition of NF-kappa B activity through induction of I kappa B synthesis. Science 270(5234): 286-290 https://doi.org/10.1126/science.270.5234.286
  10. Baldwin, A.S., Jr. (1996). The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu. Rev. Immunol. 14: 649-683 https://doi.org/10.1146/annurev.immunol.14.1.649
  11. Birchall, N., Or10w, S.J., Kupper, T. and Pawelek, J (1991). Interactions between ultraviolet light and interleukin-1 on MSH binding in both mouse melanoma and human squamous carcinoma cells. Biochem. Biophys. Res. Commun. 175(3): 839-845 https://doi.org/10.1016/0006-291X(91)91641-O
  12. Wintzen, M., Yaar, M., Burbach, J.P and Gilchrest, B.A. (1996). Proopiomelanocortin gene product regulation in keratinocytes. J. Invest. Dermatol. 106(4): 673-678 https://doi.org/10.1111/1523-1747.ep12345496
  13. Kadono, S., Manaka, I., Kawashima, M., Kobayashi, T. and Imokawa, G. (2001). The role of the epidermal endothelin cascade in the hyperpigmentation mechanism of lentigo senilis. J Invest. Dermatol. 116(4): 571-577 https://doi.org/10.1046/j.1523-1747.2001.01296.x
  14. Gilchrest, B.A., Park, H.Y., Eller, M.S. and Yaar, M. (1996). Mechanisms of ultraviolet light-induced pigmentation. Photochem. Photobiol. 63(1): 1-10 https://doi.org/10.1111/j.1751-1097.1996.tb02988.x
  15. Moon, K.Y., Ahn, K.S., Lee, J. and Kim, Y.S. (2001). Kojic acid, a potential inhibitor of NF-kappaB activation in transfectant human HaCaT and SCC-13 cells. Arch. Phann. Res. 24(4): 307-311 https://doi.org/10.1007/BF02975097
  16. Moon, K.Y., Hahn, B.S., Lee, J. and Kim, Y.S. (2001). A cell-based assay system for monitoring NF-kappaB activity in human HaCat transfectant cells. Anal. Biochem. 292(1): 17-21 https://doi.org/10.1006/abio.2001.5059
  17. Ahn, K.S., Moon, K.Y., Lee, J. and Kim, Y.S. (2003). Downregulation of NF-KB activation in human keratinocytes by melanogenic inhibitors. J. Derm. Sci. 31: 193-201 https://doi.org/10.1016/S0923-1811(03)00039-2
  18. Chung, J. H., Youn, S. H. and Koh, W.S. (1996). Ultraviolet B irradiation-enhanced interleukin (IL)-6 production and mRNA expression are mediated by IL-1 alpha in cultured human keratinocytes. J. Invest. Dermatol. 106: 715-720 https://doi.org/10.1111/1523-1747.ep12345608
  19. Boukamp, P, Petrussevska, R. T., Breitkreutz, D., Homung, J, Markham, A and Fusenig, N. E. (1998). Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell. BioI. 106(3): 761-771
  20. Na, M.K., An, R.B., Lee, S.M., Hong, N.D., Yoo, J.K, Lee, C.B., Kim, J.P. and Bae, K.H. (2001). Screening of crude drugs for antioxidative activity.kor: J Pharmacogn. 32(2): 108-115
  21. 육창수(1989). 원색한국약용식물도감, 272, 아카데미서적, 서울
  22. 신국현, 정한숙, 조선생(1995). 생열귀나무 열매의 비타민함량. 한국약용 작물학회지, 3: 21-24
  23. 우원홍, 김정훈, 문연자, 이성원, 이승연, 박정숙(2002). 황기 부탄을 분휙물이 생위의 세포성 면역기능에 미치는 영향. 약학회지, 46(1): 52-57