Biological Activities of Methanol Extract of Angelica gigas Nakai

참당귀 Methanol 추출물의 생리활성

  • Park, Kyung-Wuk (Department of Food and Nutrition, Sunchon National University) ;
  • Choi, Sa-Ra (Department of Food and Nutrition, Sunchon National University) ;
  • Hong, Hye-Ran (Department of Food and Nutrition, Sunchon National University) ;
  • Kim, Jae-Yong (Department of Food Science and Technology, Kyungpook National University) ;
  • Shon, Mi-Yae (Department of Food and Nutrition, Gyeongsang National University) ;
  • Seo, Kwon-Il (Department of Food and Nutrition, Sunchon National University)
  • 박경욱 (순천대학교 식품영양학과) ;
  • 최사라 (순천대학교 식품영양학과) ;
  • 홍혜란 (순천대학교 식품영양학과) ;
  • 김재용 (경북대학교 식품공학과) ;
  • 손미예 (경상대학교 식품영양학과) ;
  • 서권일 (순천대학교 식품영양학과)
  • Published : 2007.12.31

Abstract

The biological activities of methanol extracts of Angelica gigas Nakai, such as antioxidation, anticancer and immuno-activity, were investigated in relation to development of functional foods. Anti-oxidation activity in the methanol extracts were assessed by hydrogen donating activity, reducing power and hydroxyl radical scavenging activity. Activities were dose-dependent over concentrations of 0.1, 0.5 and 1 mg/mL, with thehydrogen donating activity being over 50% at 1 mg/mL concentration. The methanol extracts inhibited the proliferation of SW480 cells in a dose-dependent manner, and chromatin condensation and apoptotic bodies were observed by fluorescence microcopy in the cells treated with the extracts for 24 hr. Caspase-3 activity was also increased in a dose-dependent manner in cells treated with the extracts relative to control cells. The extracts did not induce the proliferation of mouse spleen cells or NO production in macrophage cells (RAW 264.7). These results show that the methanol extract had slight anti-oxidative activity and did not increase immuno-activity, but inhibited proliferation of SW480 through apoptosis via a caspase dependent pathway.

참당귀(Angelica gigas Nakai)를 기능성 식품소재 및 의약품의 원료로 활용하기 위하여 참당귀 methanol 추출물에 대한 항산화, 항암 및 면역활성을 조사한 결과는 다음과 같다. 참당귀의 항산화력을 알아보기 위하여 0.1, 0.5 및 1 mg/mL의 농도로 당귀의 methanol 추출물을 처리한 후 수소 공여능, 환원력 및 hydroxyl radical 활성을 측정하였다. 그 활성은 모두 추출물의 농도에 의존적으로 높게 나타났으며, 특히 수소공여능은 1 mg/mL에서 대조군과 비교했을때 50% 정도의 활성을 나타내었다. 참당귀 methanol 추출물을 SW480세포에 처리한 결과 대조군에 비하여 농도 의존적으로 암세포의 성장을 억제하였으며, 암세포에 참당귀 methanol 추출물을 24시간 처리하여 hoechst 염색한 후 형광현미경으로 관찰한 결과 핵의 응축 및 DNA 분절이 관찰되었다. 또한 참당귀 methanol 추출물이 처리된 암세포에서는 대조군에 비하여 농도에 의존적으로 caspase-3의 활성이 증가하였다. 참당귀의 methanol 추출물은 생쥐의 비장으로부터 분리한 면역세포에 대하여 대조군과 비교하여 상이한 차이를 보이지 않았으며, 또한 대식세포인 RAW 264.7에서도 $NO_2$(Nitrite)를 함량이 증가되지 않았다. 따라서 이들 결과는 참당귀 methanol 추출물은 약한 항산화 활성을 가지며, 면역 활성에는 큰 관련이 없지만, SW480 암 세포에서 caspase dependent pathway에 의한 apoptosis를 유도함으로서 암세포의 사멸하는 것으로 판단되어진다.

Keywords

References

  1. Park, J.H., Lee, Y.J. and Keon, S.J. (2005) Pharmacognostical studies on the dang gui from korea. Korean J. Pharmacogn., 36, 141-144
  2. Song, S.H., Seo, B.I., Kim, H.K. and Park, J.H. (2004) The effects of angelicae gigantis radix extract on hydrocortisone acetate induced model of blood stasis. Korean J. Herbology., 19, 13-21
  3. Yamada, H. (1992) Pharmacological and clinical effects of angelicae radix. 現代東洋醫學 Medical publication center Tokyo Japan., 13, 102-109
  4. Kano, Y. (1981) Physiological actions of angelicae and crude. 現代東洋醫學., 2, 43-48
  5. 陸昌洙 (1982) 漢藥의 藥理成分 臨床應用. p.738
  6. Ahn, K.S., Sim, W.S. and Kim, I.H. (1995) Decursin a cytotoxic agent and protein kinase c activator from the root of angelica gigas. Planta Med., 62, 7-9 https://doi.org/10.1055/s-2006-957785
  7. Lee, S.L. (1994) Phytology, Young Lim Pubishing Co. Seoul, p.578-580
  8. Dakaki, K. (1982) Whahanyakmoolhak, Namsandang, Tokyo, Japan, p.293-297
  9. Oh, H.S. (2001) Comparative studies of the angiogenic activity of extract of angelica gigas, A. sinensis and A. acutiloba. Ph.D. dissertation, Kyunghee University, Seoul, Korea. p.34-35
  10. Kim, K.Y. (1990) Studies on the determination of decursin in angelica gigantis radix and some umbelliferae plants. M.S Thesis, Korea Kyunghee University. p.10-21
  11. Hwang, Y.S,. Won, D.H,. Yoon, T.B. and No, H.W (1984) Studies on quality control method of crude drug preparations. Report of NIH Korea, p.341-348
  12. Ryu, K.S,. Hong, N.D,. Kim, N.J. and Kong, Y.Y. (1990) Studies on the coumarin constituents of the root of angelica gigas nakai isolation of decursinol angelate assay of decursinol angelate and decursin. Korea J. Pharmacogn., 21, 64-68
  13. Seong, N.S,. Lee, S.W,. Kim, K.S. and Lee, S.T. (1993) Environmental variation of decursin content in angelica gigas. Korean J. Crop Sci., 38, 60-65
  14. Lee, W.J,. Yoon, J.R,. Kim, E.K. and Ahn, K.T. (2000) Preparation and physicochemical properties of extracts from angelica gigantis radix of jin bu area. J. of East Coastal Research., 11, 13-22
  15. Yee, S.T,. Jeong, Y.R,. Ha, M.H,. Byun, M.W. and Jo, S.K. (1998) Effect of angelica gigantis radix-water extract on nitric oxide synthesis in mouse macrophage. J. Life Resources & Industry., 3, 1-8
  16. Rho, T.C., Choi, H.C., Lee, S.W., Kim, Y.H., Rho, M,C., Kim, Y.K. and Lee, H.S. (2001) Inhibition of nitric oxide synthesis by coumarins from ploygonum cuspidatum in lps-activated raw 264.7 cells. Korean J. Pharmacogn., 32, 181-188
  17. Blois, M.S. (1958) Antioxidant determination by the use of astable free radical. Nature., 181, 1199-1204 https://doi.org/10.1038/1811199a0
  18. Yildirim, A., Mavi, A. and Kara, A.A. (2001) Determination of antioxidant and anti microbial activities of rumex of aerobic life. Biochem. Symp., 61, 1-34
  19. Aruoma, O.I., Grootveld, M. and Halliwell, B. (1987) The role of iron in ascorbate-dependant deoxyribose degradation. Journal of Inorganic Biochemistry., 29, 289-299 https://doi.org/10.1016/0162-0134(87)80035-1
  20. Moon, S.O., Lee, J.H. and Kim, T.J. (1998) Changes in the express of c-myc, RB and tyrosine-phosphorylated proteins during proliferation of NIH 3T3 cells induced by hyaluronic acid. Experimental and Molecular., 30, 29-33 https://doi.org/10.1038/emm.1998.4
  21. Ricote, M., Garcia-Tunon, I., Fraile, B., Fernandez, C., Aller, P., Paniagual, R. and Royuelal, M. (2006) P38 MAPK protects against TNF-${\alpha}$-provoked apoptosis in LNCaP prostatic cancer cells. An International Journal on Programmed Cell Death.., 11, 1969-1975 https://doi.org/10.1007/s10495-006-0086-9
  22. Kuo, P.L., Hsu Y.L., Chang C.H. and Lin C.C. (2005) The mechanism of ellipticine-induced apoptosis and cell cycle arrest in human breast MCF-7 cancer cells. Cancer Letters., 223, 293-301 https://doi.org/10.1016/j.canlet.2004.09.046
  23. Promega Protocol. (2001) Cell titer 96${\circledr}$ aqueous one solution cell proliferation assay. Promega, USA
  24. Yee, S.T., Jeong, Y.R., Ha, M.H., Kim, S.H., Byun, M.W. and Jo, S.K. (2000) Induction of nitric oxide and TNF-${\alpha}$ by herbal plant extract in mouse macrophage. Korean Soc Food Sci Nutr., 29, 342-348
  25. Lee, S.H., Lee, Y.S., Jung, S.H., Shin, K.H., Kim, B.K. and Kang, S.S. (2003) Antioxidant activities of decursinol angelate and decursin from angelica gigas roots. Nat. Prod. Sci., 9, 170-173
  26. Kang, S.A., Han, J.A., Jang, K.H. and Choue R.W. (2004) DPPH radical scavenger activity and antioxidant effects of cham-dang-gui (angelica gigas). Korean J. Soc Food Sci Nutr., 33(7), 1112-1118 https://doi.org/10.3746/jkfn.2004.33.7.1112
  27. Kim, E.Y., Baik, I.H., Kim, J.H., Kim, S.R. and Rhyu, M.R. (2004) Screening of the antioxidant activity of some medicinal plants. Korean J. Food SCI. Technol., 36, 333-338
  28. Yim, D., Singh, R.P., Agarwal, C., Lee, S., Chi, H. and Agarwal, R. (2005) A novel anticancer agents, decursin, induces G1 arrest and apoptosis in human prostate carcinoma cells. Cancer Res., 65, 1035-44
  29. Park, S.Y,. Moon, E.Y,. Zee, O.P, and Park, E.K. (1990) Immunomodulating activity of water extract from herbal Drug ( I ) - Immunosuppressing activity of water extract from xanthii strumarium. Korean J. Immunology., 12, 119-130
  30. Lee, Y.S. (1999) Effect of methanol extract of wild plants on mouse spleenocytes proliferation. Korea Sookmyung University the Department of Food and Nutr., a master's thesis for a degree
  31. Ko, K.S. and Kim, Y.S. (1991) An illustrated book of the korean flora. Academy Publishing Co, Korea., p.433-434
  32. Kang, Y.G., Lee, J.H., Chae, H.J., Kim, D.H., Lee, S.H. and Park, S.Y. (2003) HPLC analysis and extraction methods of decursin and decursinol angelate in angelica gigas root. Korean J. Pharmacogn., 34, 201-205
  33. Ryu, K.S., Hong, N.D., Kim, N.J. and Kong, Y.Y. (1990) Studies on the coumarin constituents of the root of angelica gigas nakai. Isolation of decursinol angelate and assay of decursinol angelate and decursin. Korean J. Pharmacogn., 21, 64-68
  34. Chi, H.J. and Kim, H.S. (1988) Studies on essential oil of plants of angelica genus in Korea (I) essential oils of angelica gigantis radix. Korean J. Pharmacogn., 19, 239-247
  35. Kim, H.S., Park, H.J. and Chi, H.J. (1980) A study of effects of the root components of angelica gigas nakai on voluntary activity in mice. Korean J. Pharmacogn., 11, 11-14
  36. Ahn, K.S., Sim, W.S. and Kim, I.H. (1996) A cytotoxic agent and protein kinase c activator from the root of angelica gigas. Planta Med., 62, 7-9 https://doi.org/10.1055/s-2006-957785
  37. Han, S.B., Kim, Y.H., Lee, C.W., Park, S.M., Lee, H.Y., Ahn, K.S., Kim, I.H. and Kim, H.M. (1998) Charateristic immunostimulation by angelan isolated from angelica gigas nakai. Immunopharmacol., 40, 39-48 https://doi.org/10.1016/S0162-3109(98)00026-5
  38. Salikhova, R.A. and Poroshenko, G.G. (1995) Antimutagenic properties of medicinal angelica (angelica archangelica L.) studied with the micronucleus tes. ross akad. Med. Nauk., 1, 58-61
  39. Kim, H.H., Ahn, K.S., Han, H., Choung, S.Y., Choi, S.Y. and Kim, I.H. (2005) Decursin and PDBu; two PKC activators distinctively acting in the megakaryocytic differentiation of K562 human erythroleukemia cells. Leukemia Research., 29, 1407-1413 https://doi.org/10.1016/j.leukres.2005.05.001
  40. Kim, H.H., Bang, S.S., Choi, J.S., Han, H.G. and Kim, I.H. (2005) Involvement of PKC and ROS in the cytotoxic mechanism of anti-leukemic decursin and its derivatives and their structure-activity relationship in human K562 erythroleukemia and U937 myeloleukemia cells. Cancer Lett., 223, 191-201 https://doi.org/10.1016/j.canlet.2004.10.025
  41. Hong, M.H. (1972) Statistical studies on the formularies of oriental medicine (I). prescription frequency and their origin distribution of herb drugs. Korean J. Pharmacogn., 3, 57-64