Journal of Life Science (생명과학회지)

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

DOI QR Code

The Effects of Medicinal Herbs Extracts on Estrogen-like Activities and Osteoblast Proliferation and Differentiation

한약재 추출물의 에스트로겐 유사활성 및 조골세포 증식과 분화에 미치는 영향

  • Kim, Mihyang (Department of Food and Nutrition, Silla University) ;
  • Kim, Bokyung (Department of Food and Nutrition, Silla University) ;
  • Kim, Jae-Deog (Department of Green-Chemistry Convergence Engineering, Silla University) ;
  • Kang, A-Ram (Department of Food and Nutrition, Silla University) ;
  • Lee, Chang-Eun (Department of Green-Chemistry Convergence Engineering, Silla University) ;
  • Seo, Jungmin (Department of Food and Nutrition, Silla University) ;
  • Lee, Dong-Geun (Department of Bioscience, Graduate School, Silla University) ;
  • Jo, Jung-Kwon (Department of Bioscience, Graduate School, Silla University) ;
  • Kim, Yuck Yong (ISAAC F&B Co., Marine Bio-industry Department Center) ;
  • Yu, Ki Hwan (ISAAC F&B Co., Marine Bio-industry Department Center) ;
  • Lee, Sang-Hyeon (Department of Green-Chemistry Convergence Engineering, Silla University)
  • 김미향 (신라대학교 의생명과학대학 식품영양학과) ;
  • 김보경 (신라대학교 의생명과학대학 식품영양학과) ;
  • 김재덕 (신라대학교 일반대학원 그린화학융합공학과) ;
  • 강아람 (신라대학교 의생명과학대학 식품영양학과) ;
  • 이창은 (신라대학교 일반대학원 그린화학융합공학과) ;
  • 서정민 (신라대학교 의생명과학대학 식품영양학과) ;
  • 이동근 (신라대학교 일반대학원 바이오과학과) ;
  • 조정권 (신라대학교 일반대학원 바이오과학과) ;
  • 김육용 ((주)이삭에프앤비) ;
  • 유기환 ((주)이삭에프앤비) ;
  • 이상현 (신라대학교 일반대학원 그린화학융합공학과)
  • Received : 2017.02.08
  • Accepted : 2017.02.24
  • Published : 2017.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

The purpose of this study was to investigate the effect of 3 types of medicinal herbs (Glycyrrhizae radix, Astragali radix and Dioscorea rhizoma) extracts on estrogen-like activities, proliferation and differentiation in osteoblast. Human breast cancer cell line MCF7 was transfected using an estrogen responsive luciferase reporter plasmid for measure the estrogen-like activity. Estrogen-like activities of extracts were in the range of 1.11~5.73 fold to that of negative control. The extract of G. radix showed the strongest estrogen-like activities. The estrogen-like activities of 50 and $500{\mu}g/ml$ extracts of G. radix were similar to that of $10^{-8}$ and $10^{-7}$ M standard solution ($17{\beta}-estradiol$), respectively. G. radix extract showed no cytotoxicity against osteoblast MC3T3-E1 cells at $1{\sim}1,000{\mu}g/ml$. The extract of A. radix showed no significant proliferation of osteoblast. However, the extract of G. radix and D. rhizome showed maximum 148% and 133% proliferation effects. The extract of G. radix also increased alkaline phosphatase activity and the maximum was 122% at $100{\mu}g/ml$ compared to that of control. The nodule formation by the method of the Alizarin red S staining increased compared to control. These results suggest that G. radix is able to perform the bone formation and prevent osteoporosis.

본 연구는 3종의 한약재(감초(Glycyrrhizae radix), 황기(Astragali radix) 및 산약(Dioscorea rhizome)) 추출물의 에스트로겐 유사활성 및 조골세포 증식과 분화에 미치는 영향을 검토하고자 실험을 진행하였다. 에스트로겐 유사활성 측정을 위하여 인체 유방암 세포주인 MCF7 세포를 luciferase 유전자를 리포터로 사용하는 에스트로겐 반응성 리포터 시스템을 가지는 세포로 형질전환하여 사용하였다. 상기 세포를 이용하여 추출물의 에스트로겐 유사활성을 측정한 결과, 한약재 추출물이 음성대조군과 비교하여 1.11배~5.73배 높은 에스트로겐 유사활성을 나타내었다. 그 중 감초 추출물이 가장 높은 에스트로겐 유사활성을 보였다. 감초 추출물의 에스트로겐 유사활성은 추출물의 농도가 $50{\mu}g/ml$ 및 $500{\mu}g/ml$ 일 때 각각 $10^{-8}M$ 및 $10^{-7}M$ $17{\beta}-estradiol$과 거의 유사한 활성을 나타내었다. 조골세포주인 MC3T3-E1 세포를 이용한 실험에서는 감초 추출물의 농도가 $1{\sim}1,000{\mu}g/ml$ 범위일 때 독성을 나타내지 않았다. 황기 추출물은 조골세포에 있어서 유의적인 세포 증식률을 보이지 않았다. 그러나, 감초 및 산약 추출물은 각각 148% 및 133%의 최대 증식률을 보였다. 또한, 감초 추출물은 대조군과 비교하여 alkaline phosphatase(ALP) 활성이 증가하였으며, 추출물의 농도가 $100{\mu}g/ml$ 일 때 최대 122%의 ALP 활성을 나타내었다. 또한, Alizarin red S 염색법을 이용한 석회화 형성능 측정에서도 대조군과 비교하여 석회화가 증가하였다. 이러한 결과로 미루어 보아 감초 추출물이 골 형성 및 골다공증 예방 효과가 우수한 식품 소재인 것으로 사료된다.

Keywords

References

  1. Cho, S. H. 1995. The electron microscopic observation of the effects of estrogen on the osteoblast-like cell (Ros-17/2.8-5) differentiation. J. Kor. Soc. Menopause 1, 28-34.
  2. Choi, H. 2008. Development and conservation of the female skeleton. Osteoporosis 6, 1-7.
  3. Choi, S. Y. 2002. Effects of irradiation on the mRNA expression of the type I collagen and alkaline phosphatase in MC3T3-E1 osteoblastic cell line. Ph.D. dissertation, Chonbuk University, Jeonju, Korea.
  4. Choi, W. H. 2002. Medical treatment of osteoporosis: Hormone replacement therapy. Hanyang J. Med. 22, 19-25.
  5. Choi, G. Y. and Kim, B. W. 2010. Experimental study on the antioxidant and antimicrobial properties of Dioscoreae Rhizoma. Kor. J. Orient. Int. Med. 31, 290-297.
  6. Davis, S. R., Murkies, A. L. and Wilcox, G. 1998. Phytoestrogens in clinical practice. Integr. Med. 1, 27-34. https://doi.org/10.1016/S1096-2190(98)00019-5
  7. Declercq, H. A., Verbeeck, R. M., De Ridder, L. I., Schacht, E. H. and Cornelissen, M. J. 2005. Calcification as an indicator of osteoinductive capacity of biomaterials in osteoblastic cell cultures. Biomaterials 26, 4964-4974. https://doi.org/10.1016/j.biomaterials.2005.01.025
  8. Edwards, M. W., Bain, S. D., Bailey, M. C., Lantry, M. M. and Howard, G. A. 1992. $17{\beta}$ Estradiol stimulation of endosteal bone formation in the ovariectomized mouse: an animal model for the evaluation of bone-targeted estrogens. Bone 13, 29-34. https://doi.org/10.1016/8756-3282(92)90358-4
  9. Gerbarg, P. L. and Brown, R. P. 2016. Pause menopause with Rhodiola rosea, a natural selective estrogen receptor modulator. Phytomedicine 23, 763-769. https://doi.org/10.1016/j.phymed.2015.11.013
  10. Golub, E. E. and Boesze-Battaglia, K. 2007. The role of alkaline phosphatase in mineralization. Curr. Opin. Orthop. 18, 444-448. https://doi.org/10.1097/BCO.0b013e3282630851
  11. Green, L. M., Reade, J. L. and Ware, C. F. 1984. Rapid colormetric assay for cell viability: application to the quantitation of cytotoxic and growth inhibitory lymphokines. J. Immunol. Methods 70, 257-268. https://doi.org/10.1016/0022-1759(84)90190-X
  12. Ha, J. M. and Lee, S. H. 2003. Verification of estrogenic activity in ethanol extracts of marine organisms using in vitro test system. J. Life Sci. 13, 799-804. https://doi.org/10.5352/JLS.2003.13.6.799
  13. Han, S. B., Gu, H. A., Kim, S. J., Kim, H. J., Kwon, S. S., Kim, H. S., Jeon, S. H., Hwang, J. P. and Park, S. N. 2013. Comparative study on antioxidative activity of Glycyrrhiza uralensis and Glycyrrhiza glabra extracts by country of origin. J. Soc. Cosmet. Sci. Korea 39, 1-8.
  14. Jang, M. K., Lee, O. H., Kim, M. H., Lee, Y. A., Kang, S. R. and Lee, S. H. 2008. Evaluation of the estrogenic activities of natural products. J. Clin. Biochem. Nutr. 43, 145-148.
  15. Jeon, M. J., Kim, S., Kim, B. K., Cheon, J., Park, S. H., Oh, E., Lee, S. H. and Kim, M. 2013. The effects of seaweed gongjindan on estrogen like activities, platelet aggregation and serum lipid levels in ovariectomized rats. J. Life Sci. 23, 1155-1162. https://doi.org/10.5352/JLS.2013.23.9.1155
  16. Jung, C. S. 2012. Aging society and labor market. J. Digit. Converg. 10, 185-194.
  17. Jung, M. 2008. The effect of hormone therapy on bone. J. Kor. Soc. Osteoporosis 6, 71-78.
  18. Kim E. J. 2001. Antioxidative effects and their active constituents of Astragali Radix. Ph.D. dissertation, Sookmyung Women's University, Seoul, Korea.
  19. Kim, D. H. 2011. Quality characteristics of fish paste prepared with Astragalus membranaceus powder. Food Eng. Prog. 15, 362-369.
  20. Kim, H. J., Jang, H. N., Bae, J. Y., Ha, J. H. and Park, S. N. 2014. Antimicrobial activity, quantification and bactericidal activities of licorice active ingredients. Kor. J. Microbiol. Biotechnol. 42, 386-392. https://doi.org/10.4014/kjmb.1410.10002
  21. Kim, N. Y., Jang, H. J., Lee, D. G., Jang, M. K., Lee, S. W., Jeon, M. J., Kim, M. H., Kim, S. G. and Lee, S. H. 2011. Establishment of in vitro detection system for iNOS expression and the verification of suppressive effect by pine needle extract. Kor. Soc. Biotechnol. Bioeng. J. 26, 172-176.
  22. Kim, T. H., Lee, H. H., Chung, S. H. and Park, H. S. 2011. Differentiation in the management of osteoporosis between premenopausal and menopausal women. J. Kor. Soc. Menopause 17, 21-26.
  23. Kinto, N., Iwamoto, M., Enomoto-Iwamoto, M., Noji, S., Ohuchi, H., Yoshioka, H., Kataoka, H., Wada, Y., Yahao, G., Takahashi, H. E., Yoshiki, S. and Yamaguchi, A. 1997. Fibroblasts expressing sonic hedgehog induce osteoblast differentiation and ectopic bone formation. FEBS Lett. 404, 319-323. https://doi.org/10.1016/S0014-5793(97)00014-8
  24. Lee, Y. G., Song, J., Lee, S. B., Kim, K. N. and Choi, S. H. 2003. Influence of calcium phosphate glass for periodontal regeneration on MC3T3-E1 proliferation, differentiation and calcification. J. Kor. Res. Soc. Dent. Mater. 30, 181-188.
  25. Lee, S. H. 2003. Verification of estrogenic activities in ethanol extracts of oriental herbal medicines using in vitro detection system. Kor. J. Orient. Physiol. Pathol. 17, 1054-1058.
  26. Lee, S. H., Jung, B. H., Kim, S. Y. and Chung, B. C. 2004. Determination of phytoestrogens in traditional medicinal herbs using gas chromatography-mass spectrometry. J. Nutr. Biochem. 15, 452-460. https://doi.org/10.1016/j.jnutbio.2004.01.007
  27. Lissin, L. W. and Cooke, J. P. 2000. Phytoestrogens and cardiovascular health. J. Am. Coll. Cardiol. 35, 1403-1410. https://doi.org/10.1016/S0735-1097(00)00590-8
  28. McAnuff, M. A., Harding, W. W., Omoruyi, F. O., Jacobs, H., Morrison, E. Y. and Asemota, H. N. 2005. Hypoglycemic effects of steroidal sapogenins isolated from Jamaican bitter yam, Dioscorea polygonoides. Food Chem. Toxicol. 43, 1667- 1672. https://doi.org/10.1016/j.fct.2005.05.008
  29. Jeon, M. H. and Kim, M. H. 2011. Effect of Hijikia fusiforme fractions on proliferation and differentiation in osteoblastic MC3T3-E1 cells. J. Life Sci. 21, 300-308. https://doi.org/10.5352/JLS.2011.21.2.300
  30. Mori, Y., Hiraki, Y., Shukunami, C., Kakudo, S., Shiokawa, M., Kagoshima, M., Mano, H., Hakeda, Y., Kurokawa, T., Suzuki, F. and Kumegawa, M. 1997. Stimulation of osteoblast proliferation by the cartilage derived growth promoting factors chondromodulin‐I and‐II. FEBS Lett. 406, 310-314. https://doi.org/10.1016/S0014-5793(97)00291-3
  31. Park, S. H., Jeon, M. J., Jang, M. K., Lee, S. J., Kim, B. K., Jeon, M. J., Kim, S. Y., Kim, M., Lee, D. G., Lee, T. H., Nam, J. S., Hur, J. W. and Lee, S. H. 2013. Verification of estrogen like activities of herbal medicines using an in vitro detection system. Kor. J. Orient. Physiol. Pathol. 27, 752-758.
  32. Park, Y. M., Lim, J. H. and Seo, E. W. 2015. Effect of Dioscorea Rhizoma ongastritis by acute gastric ulcer in rats. Kor. J. Plant Res. 28, 1-8. https://doi.org/10.7732/kjpr.2015.28.1.001
  33. Jeong, Y. and Lee, Y. 2009. Protective effect of Rhizoma dioscoreae on the gastric mucosal lesions Induced by alcohol in rats. Kor. J. Orient. Physiol. Pathol. 23, 639-644.
  34. Yun, J. 2010. Effects of Chrysanthemum indicum L. extract on the activity and differentiation of osteoblastic MC3T3-E1 cells. M.S. Thesis. dissertation, Duksung women's University, Seoul, Korea.
  35. Yun, J. H., Hwang, E. S. and Kim, G. H. 2011. Effects of Chrysanthemum indicum L. extract on the growth and differentiation of osteoblastic MC3T3-E1 Cells. J. Kor. Soc. Food Sci. Nutr. 40, 1384-1390. https://doi.org/10.3746/jkfn.2011.40.10.1384