Antiproliferative and Antioxidative Activities of Methanol Extracts of Echinacea angustifolia

Echinacea angustifolia 메탄올 추출물의 암세포 증식억제 및 항산화 효과

  • Lee Joon-Kyoung (Department of Food & Nutrition, Kyunghee University) ;
  • Koo Seung-Ja (Department of Food & Nutrition, Kyunghee University)
  • 이준경 (경희대학교 식품영양학과) ;
  • 구성자 (경희대학교 식품영양학과)
  • Published : 2005.06.01

Abstract

Echinacea, also blown as the purple coneflower, is a herbal medicine that has been used for centuries, customarily as a treatment for the common cold, coughs, bronchitis, upper respiratory infections, and some inflammatory conditions. We investigated the effects of methanol extracts of Echinacea angustifolia on the cytotoxicity against cancer cells $(HepG_2,\;3LL,\;HL60,\;L1210)$ and antioxidative activity. From the test results, each part of Echinaceashowed a cytotoxic effect against the cancer cell lines, and this cytotoxic effect increased with increasing sample concentration. At 1.0 mg/mL concentration the relative cytotoxic activities of the flower bud, leaf, stern and root parts were $90.5\%,\;52.7\%,\;37.1\%\;and\;19.2\%$, respectively, in $HepG_2$ cells, and $75.5\%,\;93.3\%,\;81.2\%,\;and\;75.1\%$ respectively, in HL60 cells, as evaluated by MTT assay. $IC_{50}(50\%\;inhibitory\;concentration)$ of the methanol extracts of the Echinacea flower bud was 0.214 mg/mL on /$HepG_2$ cells, and that of the Echinacea leaf and root was 0.166 mg/mL and 0.210 mg/mL, respectively, on HL60 cells. After /$HepG_2$ cells were incubated for 6 days at $37^{\circ}C$ with various concentrations of each part, the cell number increased while the inhibition rate on the /$HepG_2$ cell growth decreased. The antioxidative activities of the flower bud, leaf, stem and root parts were $59.0\%$ (0.75 mg/mL), $80.76\%$ (0.5 mg/mL), $95.5\%$ (0.25mg/mL) and $98.15\%$ (0.25 mg/mL), respectively, as evaluated by electron donating ability. These results indicated that Echinacea angustifolia has strong anticancer and antioxidative effects in vitro.

Echinacea angustifolia의 부위별(꽃봉오리, 잎, 줄기 및 뿌리) 메탄을 추출물의 암세포(HepG2, 3LL, HL60, L1210)를 대상으로 한 항암 활성과 전자공여능을 검색한 결과는 다음과 같다. 1. 에키네시아 메탄을 추출물의 간암세포인 HepG2 cell 대한 MTT assay는 농도의존적으로 세포독성 효과가 증가하였으며, 인간유래 백혈암세포인 HL60 cell의 경우에 잎과 뿌리 추출물은 저농도에서부터 독성효과가 컸고, 줄기와 꽃봉오리는 저농도에서는 독성효과가 낮았으나 고농도로 갈수록 독성효과가 커짐을 알 수 있었다 폐암세포인 3LL cell과 마우스 유래 백혈암세포인 L1210 cell에 대한 경우는 세포독성효과가 없었다. 2. Hemacytometer에 의한 HepG2 cell의 암세포 성장에 미치는 효과는 배양기간이 증가함에 따라 농도의존적으로 증식억제 효과가 증가되었다. 3. HepG2 세포주의 형태학적 변화에서 대조군은 암세포가 조밀하게 중첩되어 증식되었으나 시료를 0.5 mg/mL 이상의 농도로 첨가하였을 때 세포의 결속력이 감소되어 세포주위가 흐트러지고 세포가 사멸된 것을 관찰 할 수가 있었다. 4. 전자공여능의 수준은 부위에 따라 최고의 EDA를 나타내는 농도가 달랐으며, 뿌리와 줄기부위는 저농도에서도 매우 높은 전자공여능을 나타내었다.

Keywords

References

  1. Barbara G, Campling JP, Peter RG, Susan PCC (1988) : Use of the MTI assay for rapid determination of chemosensitivity of human leukemic blast cells. Leukemia Res., 12 : 823-831 https://doi.org/10.1016/0145-2126(88)90036-7
  2. Barrett B, Kiefer D, Rabago D (1999) : Assessing the risks and benefits of herbal medicine: An overview of scientific evidence. Altern. Ther., 5 : 40-49
  3. Burger RA, Torres AR, Warren RP, Caldwell YD, Hughes BG (1997) : Echinacea-induced cytokine production by human macrophages. Int. J. Immunopharmacol., 19 ; 371-379 https://doi.org/10.1016/S0192-0561(97)00061-1
  4. Carmichell J, Mitchell JB, DeGraff WG, Gamson J, Gazadar AF, Johnson BE, Glastein E, Minna JD (1988) : Chemosensitivity testing of human lung cancer cell lines using the MTT assay. Br. J. Cancer, 57 : 540-547
  5. Chung HJ, Noh KL (2000) ; Screening of electron donating ability, antibacterial activity and nitrite scavenging effect of some herbal extracts. Korean J. Soc. Food Sci., 16 : 372-377
  6. Doll R, Peto R (1981) : The cause of cancer; quantitative estimate of avoidable risks of cancer in the United States today. J. Natl. Cancer Inst., 66 : 1191
  7. Dorch W (1996) ; Clinical application of extracts of Echinacea purpurea or Echinacea pallida. Critical evaluation of controlled clinical studies. Z. Arzil. Fortbild., 90 : 117-122
  8. Dziedzic SZ, Hudson RJF (1983) : Hydroxy isoflavones as antioxidants for edible oil. Food Chem., 11 : 161 https://doi.org/10.1016/0308-8146(83)90099-7
  9. Facino RM, Carh M, Aldini G, Saibene L, Pietta P, Mauri P (1995) : Echinacoside and caffeoyl conjugates protect collagen from echinacea extracts in the prevention of skin photodamage. Planta Med., 61 : 510-514 https://doi.org/10.1055/s-2006-959359
  10. Foster S (1991) : Echinacea: Nature's Immune Enhancer. Healing Arts Press
  11. Franceschi RT, James WM, Zerlauth G (1985) $1{\alpah}$, 25-dihydroxy vitamin D3 specific regulation of growth, morphology and fibronection and a human osteosarcoma cell line. J. Cell Physiol., 123 : 401-409 https://doi.org/10.1002/jcp.1041230316
  12. Graisbauer M, Scheleich T, Stickl HA, Wilczek I (1990) : The effect of Echinacea purpurea Monk on phagocytosis in granulocytes measured by chemiluminescence. (In German.) Arzneimittelforchung. 40 : 594-598
  13. Hobbs C (1994) : Echinacea: a literature review. HerbalGram 30 : 34-48
  14. Hobbs C (1995) : Echinacea: The Immune Herb. Botanica Press
  15. Hyun JW, Lim KH, Shin JE, Sung MS, Won YJ, Kim YS, Kang SS, Chang IM., Woo WS, Paik WH, Kim HJ, Woo ER, Park HK and Park JG (1994) : Antineoplastic effect of extracts from traditional medicinal plants and various plants (in Korean). Kor. J. Pharmacol., 25(2) : 171-177
  16. Kang YH, Park YK, Oh SR, Moon KD (1995) : Studies on the physiological functionality of pine needle and mugwort extracts. Korean J. Food Sci. Technol., 27 : 978-984
  17. Kim JH, Kim KM, Yoo NC, Choi JH, Lim HY, Roh JK, Lee KS, Kim BS (1993) : Effects of verapamil, tamoxifen and cyclosporin A for the modulation of multidrug resistance in human lung cancer cell lines. J. Kor. Cancer Assoc., 25 : 225-230 https://doi.org/10.1007/BF00689837
  18. Lersch C, Zeuner M, Bauer A, Siebenrock K, Hart R, Wagner F, Fink U, Dancygier H, Classen M (1990) : Stimulation of the immune response in outpatients with hepatocellular carcinomas by low doses of cyclophosphamide (LDCY), Echinacea purpurea extracts (Echinacin) and thymostimulin. Arch. Geschwulsiforsch, 60 : 379-383
  19. Li TS, Wang LCH (1998) : Physiological components and health effects of ginseng, Echinacea, and sea buckthorn. In Functional Foods.: Biochemical and Processing Aspects, Maza G. Ed., Technomic Publishing, Lancaster, PA, 329-356
  20. Linert D, Anklam E, Panne U (1998) : Gas chromatographymass spectral analysis of roots of Echinacea; species and classification by multivariate data analysis. Phytochem. Anal., 88-98 https://doi.org/10.1002/(SICI)1099-1565(199803/04)9:2<88::AID-PCA384>3.0.CO;2-4
  21. Luettig B, Steinmuller C, Gifford GE, Wagner H, LohmannMatthes ML (1989) : Macrophage activation by the polysaccharide arabinogalactan isolated from plant cell cultures of Echinacea purpurea. J. Natl. Cancer Inst., 81: 669-675 https://doi.org/10.1093/jnci/81.9.669
  22. Melchart D, Linde K, Worku F, Sarkady L, Holzmann M., Jurcic K, Wagner H (1995) : Results of five randomized studies on the immuno-nomodulatory activity of preparations of Echinacea. J. Alternative Complementary Med., 1 : 145-160 https://doi.org/10.1089/acm.1995.1.145
  23. Mitsuda H, Yasumoto K, Iwaki K (1966) : Antioxidative action of linoleic acid. Nippon Eiyo Shokuryo Gakkaish, 19 : 210 https://doi.org/10.4327/jsnfs1949.19.210
  24. Pamham MJ (1996) : Benefits-risks assessment of the squeezed sap purple coneflower (Echinacea purpurea) for long-term oral immunostimulation. Phytomedicine, 3 : 95-102 https://doi.org/10.1016/S0944-7113(96)80020-1
  25. Rosier J, Steinmuller C, Kiderlen A, Emmendorffer A, Wagner H, Lohmann-Matthes ML (1991) : Application of purified polysaccharides from cell cultures of the plant Echinacea purpurea to mice mediates protection against systemic infections with Listeria monocytogenes and Candida albicans. Int. J. Immunopharmacol., 13 : 27-37 https://doi.org/10.1016/0192-0561(91)90022-Y
  26. Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH, Currens MJ, Seniff D, Bayed MR (1988) : Evaluation of a soluble tetrazolium/ formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res., 48 : 4827
  27. Se D, Broumand N, Sahl L, Tilles JG (1997) : In vitro effects of echinacea and ginseng on natural killer and antibody-dependent cell cytotoxicity in healthy subjects and chronic fatigue syndrome or accquired immunodeficiency syndrome patients. Immunopharmacology, 35 : 229-235 https://doi.org/10.1016/S0162-3109(96)00125-7
  28. Steinmuller C, Rosier J, Grottrup E, Franke G, Wagner H, Lohmann-Matthes ML (1993) : Polysaccharides isolated from plant cell cultures of Echinacea purpurea enhance the resistance of immunosupressed(?) mice against systemic infections with Candida albicans and Listeria monocytogenes. Int. J. Immunopharmacol., 15 : 605-614 https://doi.org/10.1016/0192-0561(93)90078-D
  29. Stirnpel M, Proksh A, Wagner H, Lohmann-Matthes ML (1984) : Macrophage activation and induction of macrophage cytotoxicity by purified polysaccharide fractions from the plant Echinacea purpurea. Infect. Immun., 46 : 845-849
  30. Tragni E, Galli CL, Tubaro A, Del NP, Della LR (1988) : Antiinflammatory activity of Echinacea angustifolia fractions separated on the basis of molecular weight. Pharmacol. Res. Commun., 20(suppl 5) : 87-90
  31. Wagner H, Stuppne, H, Schafer W, Zenk M (1988) : Immunologically active polysaccharides of Echinacea purpurea cell cultures. Phytochemistry, 27 : 119-126 https://doi.org/10.1016/0031-9422(88)80601-0
  32. Wagner HKM (1995) : Immunostimulants and adaptogens from plants. In Photochemistry of Medicinal Plants. Armason T, Ed., Plenum Press, New York, p 1-18