한국식품과학회지 (Korean Journal of Food Science and Technology)

한국식품과학회 (Korean Society of Food Science and Technology)
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칡의 부위별 골다공증 치료효과

Pueraria lobata Ohwi as an Osteoporosis Therapeutics

  • 김정숙 (한국한의학연구원 한약제제연구부) ;
  • 하혜경 (한국한의학연구원 한약제제연구부) ;
  • 김혜진 (한국한의학연구원 한약제제연구부) ;
  • 이제현 (경희대학교 약학대학 한약학과) ;
  • 송계용 (중앙대학교 의과대학 병리학교실)
  • 발행 : 2002.08.01
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초록

콩과 식물인 갈근에는 genistein과 daidzein이 함유된 것으로 알려졌으므로 본 연구는 갈근(PR), 갈화(PF) 및 어린순인 갈용(PL)에 함유된 phytoestrogen의 함량을 HPLC법으로 정량 분석하였다. 동시에 phytoestrogen의 골다공증 치료제에 대한 효능 검색의 일환으로 조골세포의 증식작용(MTT법 및 ALP분석)과 피골세포의 활성(TRAP법)을 측정하였고 난소절제 흰쥐에 PR 1 g/kg/day(PR-1) 및 PR 5 g/kg/day(PR-5)을 9주 동안 투여한 후 혈액분석, 혈장 Alkaline phospatase(ALP), 칼슘, 무기성 인산염, 총콜레스테롤, LDL- 및 HDL-콜레스테롤을 측정하였고 적출된 경골 및 요추골의 소주골면적을 측정하였다. PR과 PF의 총 daidzein의 함량은 $10436{\pm}2144\;mg/kg$와 $1003{\pm}206\;mg/kg$이었고 genistein과 formononetin은 PR에만 존재하고 PF 및 PL에는 함유되어 있지 않았다. 인간 유사 조골 세포주인 Saos-2에 PR은 대조군의 158% 정도(5 mg/mL)의 세포증식효과와 ALP 활성을 증가시켰으나 전처리한 PR, PF 및 PL의 세포증식효과는 없었고 PL만이 ALP 활성을 증가시켰다. 파골세포의 증식억제실험에서 genistein의 $IC_{50}$는 $1.57{\times}10^{-4}\;mg/mL$ $(5.81{\times}10^{-7}\;M)$이었고, daidzein과 PR은 효과가 없었다. 난소절제 흰쥐에 대한 in vivo 실험에서 대조군이 sham군보다 난소적출 1주 후부터 체중의 증가가 급격하게 나타났으나 PR의 투여로 둔화되었다. PR 투약에 의한 자궁의 무게는 PR-5군은 대조군에 비해 증가되었고(p<0.05), PR-1군은 대조군과 유의성이 없었다. 혈장 ALP의 활성은 주령의 증가에 EK라 감소하는 경향을 보였고, HDL-콜레스테롤의 농도는 모든 군에서 주령에 따라 감소되었으나(p<0.01), PR-5군의 LDL-콜레스테롤의 농도는 대조군에 비해 감소되었다(p<0.01). 대조군에 비해 sham군의 경골 및 요추골의 면적은 증가되었고(p<0.01), PR 투여군(PR-1 및 PR-5)도 sham보다 적으나 대조군에 비해 소주골 면적이 증가되었다(각 p<0.01). 특히 고용량의 PR 투여는 PR의 골다공증 치료효과가 우수함을 증명하였으며(p<0.01), PL과 PF는 효능이 미비하나 PR은 골다공증의 예방 및 치료제로는 상당히 우수한 한약재임이 확인되었다.

It is reported that Pueraria Radix contains phtoestrogens whereas flower, and bud of Pueraria lobata Ohwi were not known. In the present study, we determined the amount of phytoestrogen in each portion of P. lobata Ohwi and carried out therapeutic effects of osteoporosis. The amounts of genistein, daidzein, and formononetin in Pueraria Radix (PR), Pueraria Flos (PF), and young Pueratia Folium (PL) were quantitated using a HPLC system. Proliferation of osteoblast and growth inhibitory effect on osteoclast were measured in order to screen their effects on osteoporosis. Proliferation of osteoblast-like cells (Saos-2) was analyzed by both MTT methods and alkaline phosphatase (ALP) assays. Growth inhibitory effect on osteoclast was also detected as Tartrate resistant acid phosphatase (TRAP) assay. Ovariectomized rat as an in vivo animal model was selected and administrations of PR were 1 g/kg/day (PR-1) and 5 g/kg/day (PR-5) for 9 weeks, respectively. Trabecular bone areas (TBAs) of tibia and lumbar were analyzed usibg histomorphological methods. Results show that PR contains the highest level of daidzein ($10435{\pm}2143\;mg/kg$ of dried herb) and stimulated ALP activity, approximately 160% of the control. Growth inhibitory effect on osteoclast by both PR and daidzein were almost identical with control although $IC_{50}$ of genistein was $5.81{\times}10^{-7}$ M. Increases in body weight of OVX rats were suppressed by administration of PR but wet weights of uterus in PR-5 group were increased (p<0.05). Plasma ALP and HDL-cholesterol levels were decreased following ages (p<0.01), and LDL-cholesterol level was also decreased in PR-5 group at 20 week of age (p<0.01). TBAs of tibia and lumbar in PR-1 and PR-5 groups were higher than those of the control although the values were less than those of the sham group (each p<0.01) In conclusion, administrations of PR prevented loss of TBAs of tibia and lumber in OVX rats, while PL and PF did not (p<0.01).

키워드

참고문헌

  1. Faculities in Department of Herbology. Oriental Herbology. pp. 242-244, 167-168. Young-Lim Publishing Co., Seoul (1991)
  2. Editorial Committee. Korean Pharmacopoeia. 7th ed. pp. 721-988. Korean Medical Index Press, Seoul, Korea (1998)
  3. Pharmacopoeia Committee. DPRK Pharmacopoeia, p. 265. Medical Science Press, Pyungyang, The Democratic People's Republic of Korea(1995)
  4. Editorial. Committee. Chinese Pharmacopoeia, pp. 16-17. Guang-dong Science Press, Guang-dong, China (1995)
  5. Editorial Committee. Japanese Pharmacopoeia. 13th ed. pp. 2277-2279. Hirogawa Press, Tokyo, Japan (1996)
  6. Kurihara, T. and Kikuchi, M. Studies on the constituents of flowers. IV. On the components of the flower of Pueraria thubergiana Benth. (3). Yakugaku Zasshi. 96: 1486-1488 (1976) https://doi.org/10.1248/yakushi1947.96.12_1486
  7. Lee, K.-T., Sohn, I.-C., Kong, E.-A., Kim, D.-H., Choi, S.-K., Choi, J.-W. and Park, H.-J. Antioxidative and cytoprotective effects of isoflavones isolated from Pueraria thunbersiana flowers. Yakhak Hoeji 43: 736-742 (1999)
  8. Kinjo, J., Takeshita, T., Abe, Y., Terada, N., Yamashita, H., Yamasaki, M., Takeuchi, K., Murasaki, K., Tomimatsu, T. and Nohara, T. Studies on the constituents of Pueraria Iobata. IV. Chemical constituents in the flowers and the leaves. Chem. Pharm. Bull. 36: 1174-1179 (1988) https://doi.org/10.1248/cpb.36.1174
  9. Consensus Conference. Osteoporosis. JAMA 252: 799-802 (1984)
  10. Ryan, P.J., Evans P., Gibson. T, and Fogelman, I. Osteoporosis and chronic back pain: A study with single-photon emission computed tomography bone scintigraphy. J. Bone Miner. Res. 7: 1455-1460(1992) https://doi.org/10.1002/jbmr.5650071213
  11. Spencer, H., Rubio, N., Rubio, E., Indreika, M. and Seitam, A. Chronic alcoholism: Frequently overlooked cause of osteoporosis in men. Am. J. Med. 80: 393-397 (1986) https://doi.org/10.1016/0002-9343(86)90712-6
  12. Wang, W., Tanaka, Y., Han, E. and Higuchi, C.M. Proliferative response of mammary glandular tissue to formononetin. Nutr. Cancer23: 131-140(1995) https://doi.org/10.1080/01635589509514369
  13. Saloniemi, H., Wahala, K., Nykanen-Kurki, P., Kallela, K. and Saastamoinen, I. Phytoestrogen content and estrogenic effect of legume fodder. Proc. Soc. Exp. Biol. Med. 208: 13-17 (1995) https://doi.org/10.3181/00379727-208-43825
  14. Bingham, S.A., Atkinson, C., Liggins, J., Bluck, A. and Coward, A. Phytoestrogens: where are we now? Brit. J. Nutr. 79: 393-406 (1998) https://doi.org/10.1079/BJN19980068
  15. Evans, G.L., Bryant, H.U., Magee, D.E. and Turner, R.T. Raloxifene inhibits bone tum over and prevents further cancellous bone loss in adult ovanectomized rats with established osteopenia. Endocrinology 137: 4139-4144 (1996) https://doi.org/10.1210/en.137.10.4139
  16. Bryant, H.U. and Dere, W.H. Selective estrogen receptor modulators: an altemative to hormone replacement therapy. Proc. Soc. Exp. Bio. Med. 217: 45-52 (1998) https://doi.org/10.3181/00379727-217-44204
  17. Kuiper, G.G.J.M., Lemmen, J.G., Carlsson, B., Corton, J.C., Safe, S.H., Van Der Saag, P.T., Van Der Burg, B. and Gustafsson, J.A. Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor B. Endocnnol. 139: 4252-4263 (1998) https://doi.org/10.1210/en.139.10.4252
  18. Kim, J.S. and Kim, C. A Study on the Constituents from the roots of Astragalus membranaceus (II). Korean J. Pharmacogn. 28: 75-79 (1997)
  19. Kim, C., Lee, Y.S., Kim, J.S. and Hahn, Y. High performance liquid chromatographic analysis of isoflavones in soybean foods. Korean J. Food Sci. Technol. 32: 25-30 (2000)
  20. Carmichael, J., DeGraff, W.G., Gazdar, A.F., Minna, J.D. and Mitchell, J.B. Evaluation of a tetrazolium-based semiautomated colorimetric assay assessment of chemosensitivity testing. Cancer Res. 47: 936-942 (1987)
  21. Kassem, M., Mosekilde, L. and Eriksen, E.F. 1,25-Dihydroxyvitamin D3 potentiates fluoride-stimulated collagen type I production in cultures of human bone marrow stromal osteoblast-like cells. J. Bone Miner. Res. 8: 1453-1458 (1993)
  22. Raab, W.P. Diagnostic value of urinary enzyme determinations. Clin.Chem. 18:5-25(1972)
  23. Shioi, A., Ross, F.P. and Teitelbaum, S.L. Enrichment of generated murine osteoclasts. Calcif. Tissue Int. 55: 387-394 (1994) https://doi.org/10.1007/BF00299320
  24. Moorehead, W.R. and Briggs, H.C. 2-Amino-2-methyl-1-propanol as the alkalizing agent in an improved continuous- Qow cresol-phthalein complexone procedure for calcium in serum. Clin. Chem. 20: 1458-1460 (1974)
  25. Knoll, M.H. and Elin, R.J. Mechanism of cefoxitin and cephalothin interference with the Jaffe method for creatinine. Clin. Chem. 29: 2044-2048 (1983)
  26. Wang, J., Chen, C.C. and Osaki, S. Optimization of the phosphorus-UV reagent. Clin. Chem. 29: 1255 (1983)
  27. Tiffany, T.O., Jansen, J.M., Bur6s, C.A., Overton, J.B. and Scott, C.D. Enzymatic kinetic rate and endpoint analysis of substrate by use ofa GeMsaec fast analyzer. Clin. Chem. 18: 829-840 (1972)
  28. Kim, C., Kim, J.S., Kim, Y.T. and Song, K.Y. Effects of herbal medicine on tibia bone changes in ovariectomized rats and SAM P6. J. Phann. Soc. Korea 42: 220-228 (1998)
  29. Rosner, B. Fundamentals of Biostatisdcs. PWS-Kent Co., Boston, MA, USA (1990)
  30. Kalu, D.N. Evoluation of the pathogenesis of postmenopausal bone loss. Bone 17: 135S-144S (1995)