생명과학회지 (Journal of Life Science)

  • 제14권6호
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  • Pages.967-974
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  • 2004
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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복분자(Rubus coreanus Miquel) 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향

Effects of Rubus coreanus Miquel Extracts on the Activity and Differentiation of MC3T3-E1 Osteoblastic Cell

  • 이지원 (계명대학교 식품가공학과) ;
  • 이인선 (계명대학교 미생물자원 개발 및 산업화 연구센터)
  • Lee Ji-Won (The Center for Traditional Microorganism Resources Center, Keimyung University) ;
  • Lee In-Seon (Dept. of Food Science & Technology, Keimyung University)
  • 발행 : 2004.12.01
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초록

골조직은 골아세포, 파골세포, 골세포 등으로 구성되며, 골개조시 여러 인자가 세포증식, 분화, 활성화 및 골대사 조절에 관여한다. 이때 조골세포의 활성은 골형성에 중요하므로, 본 연구에서는 MC3T3-E1 조골세포주를 이용하여 식용자원인 복분자의 조골세포의 증식과 분화활성에 미치는 영향을 조사하였다. 복분자 추출물이 조골세포의 성장에 미치는 영향을 MTT 검색법으로 조사한 결과, 복분자 추출물 $10\;{\mu}g/mL$ 처리시 대조군과 비교하여 $142\%$ 증가하여 조골세포에 대해 높은 성장률을 보였다. 복분자 추출물이 ALP 효소 활성에 미치는 영향을 3일 간격으로 배지교환 및 시료처리를 하면서 27일 동안 배양시간에 따른 변화를 측정하여 조사하였다. 그 결과, 농도 1, 10, $100\;{\mu}g/mL$ 처리시 $100\;{\mu}g/mL$을 제외한 나머지 분획물들이 시간이 지남에 따라 ALP 활성을 증가시켰고, $10\;{\mu}g/mL$ 농도의 추출물은 27일째에 대조군에 비해 약 2.6배 이상, 양성대조군에 비해 약 1.5배 이상 ALP 활성을 증가시켰다. 복분자 추출물은 다시 ALP 효소 염색법과 Alizarin Red 염색으로 조골세포의 ALP활성유도, 분화와 석회화 형성능을 재확인하였으며 골기질 유전자의 발현의 변화도 확인하였다. 따라서 종래의 골질환에 좋다고 알려진 식품인 복분자 추출물이 양성대조군에 비해 빠르게 세포 증식과 분화를 유도하고 있어 앞으로 복분자에 대한 좀 더 깊은 분자생물학 수준 등의 구체적인 연구들과 기작연구가 지속적으로 이루어져야 할 것이라 추측된다.

The osteoblastic cell activity is important for born formation, thus, this study was performed to investigation of that the effect of edible sources, Rubus coreanus Miquel (RCM), on the proliferation and differentiation of MC3T3-E1 osteoblastic like cell. The effects of RCM extract on cell proliferation were measured by MIT assay. At 1, $10\;{\mu}g/mL$ of RCM extract treated, that were elevated of cell proliferation to 103 and $142\%$ via control, respectively. And the cell differentiation were measured as alkaline phosphatase (ALP) activity at 3, 9, 18, and 27 days. As the results, the $10\;{\mu}g/mL$ was increased ALP activity more than 2.6 times compared with control, 1.4 times via positive control at 27th day (p<0.05). The optical concentration of RC extract was rechecked by ALP staining and Alizarin Red staining for investigation of the induction of ALP activity, nodule formation by mineralization. mRNA expression analysis showed that the RCM $(10\;{\mu}g/mL)$ increased in SOX9 as well as ALP in MC3T3-E1 cells. These results suggest that RC extract was stimulates the MC3T3-E1 cell proliferation and differentiation.

키워드

참고문헌

  1. Aldercreutz, H and Mazur, W. 1996. Phyto-estrogens in relation to cancer and other human health risks. Proc. Nutr. Soc. 55, 399-417 https://doi.org/10.1079/PNS19960038
  2. Bellows, C. G., Aubin, J. E., Heersche, J. N and Antosz, M. E. 1986. Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int. 38, 143-154 https://doi.org/10.1007/BF02556874
  3. Boonen, A., Broos, P and Deqeker, J. 1997. The prevention of treatment of age-related osteoporosis in the elderly by systemic recombinant growth factor therapy (rhIGF-I or rhRGF-$\beta$):a perspective. J internal. medicine 242, 285-290 https://doi.org/10.1046/j.1365-2796.1997.00133.x
  4. Carmichael, J., Degraff, W. G and Gadzar, A. F. 1987. Evaluation of a tetrazolium based semiautomated colorimetric assay : Assessment of chemosensitivity testing. Cancer Res. 477, 936-942
  5. Centrella, M., McCartht, T. L and Canalis, E. 1987. Transforming growth Factor B is a bifunctional regulator of replication and collagen synthesis in osteoblast-enriched cell cultures from fatal rat bone. J. Biol. Chem. 262, 2869- 2874
  6. Cho, S. H. 1995. The electron microscopic observation of the effects of estrogen on the osteoblast-like cell (Ros-17/ 2.8-5) differentiation. Korean Soc. Menopause 1, 28-34
  7. Cho, S. H., Kim, K, G., Kim, S. R., Lee, J. A., Moon, H and Hwang, Y. Y., et al. 1996. The effects of 17-$\beta$ etradiol, medroxyprogesterone acetate and parathyroid hormone on the differentiation of osteoblast cell. Korean Soc. Obst. Gyn. 39, 1497-1506
  8. Choi, E. M. 2001. Effect of soybean ethanol extract on the function of osteoblastic MC3T3-E1 cells. Ph. D thesis, University of Kyunghee, Seoul, Korea
  9. Choi, E. M., Suh, K. S., Kim, Y. S., Choue, R. W and Koo, S. J. 2001. Soybean ethanol extract increases the function of osteoblastic MC3T3-E1 cells. Phytochemistry 56, 733-739 https://doi.org/10.1016/S0031-9422(00)00484-2
  10. Choi, H. H,, Kim, J. K,, Kim, S. B and Chung, C. H. 1999. Effects of Platelet - derived Growth Factor on the Activity of Osteoblastic Cells. J. Periodont. Res. 29, 785-804
  11. Choi, J. Y., Lee, B. H., Song, K. B., Park, R. W., Kim, I. S and Sohn, K. Y., et al. 1996. Expression patterns of bone-related proteins during osteoblastic differentiation in MC3T3-E1 cells. J Cellular Biochemistry 61, 609-618 https://doi.org/10.1002/(SICI)1097-4644(19960616)61:4<609::AID-JCB15>3.0.CO;2-A
  12. Choi, S. Y. 2002. Effects of irradiation on the mRNA expression of the typeⅠcollagen and alkaline phosphatase in MC3T3-E1 osteoblasti cell line. PhD thesis., Chonbuk National University
  13. Fukayama, S and Tashjian, A. H. 1990. Stimulation by parathyroid hormone of $^{45}Ca^{2+}$ uptake in osteoblast-like cells: possible involvement of alkaline phosphatase. Endocrinology 126, 1941-1949 https://doi.org/10.1210/endo-126-4-1941
  14. Green, L. M., Reade, J. L and Ware, C. F., 1984. Rapid colometric assay for cell viability : Application to the quantitation of cytotoxic and growth inhibitory lympokines. J. Immunol. Meth. 70, 257 https://doi.org/10.1016/0022-1759(84)90190-X
  15. Harrison, G., Shapiro, I.M and Golub, E.E. 1995. The phosphatidylinositol-glycolipid anchor on alkaline phosphatase facilitates mineralizaton initiation in vitro. J. Bone Miner. Res. 10, 568-573 https://doi.org/10.1002/jbmr.5650100409
  16. Karen, M. P., Carol, C. P and Lawrence, G. R. 1995. Treatment of osteoporosis. Annu. Rev. Med. 6, 249-256
  17. Keum, J. C., Kang, K. H and Kim, S. B. 2001. Effects of estrogen on the tumor necrosis factor-$\alpha$ induced apoptosis and cytokine gene expression in MC3T3-E1 osteoblast. Korean Soc. Obst. Gyn. 44, 324-336
  18. Kim, J. W., Lee, H. J., Kang, J. H., Ohk, S. H., Choi, B. K., Yoo, Y. J., Cho, K. S and Choi, S. H. 2000. The Effect of Cyclosporin A on Osteoblast in vitro. J. Periodont. Res. 30, 747-757
  19. Kim, K. W. 2001. Optimum condition on conversion of aglycones form using $\beta$-glucosidase and isoflavones extraction from soybean. Master thesis., National University of Pykyung
  20. Kim, T. J. 1996. Korean resources plants II. Publishing Department of Seoul University, Seoul, Korea
  21. Kinto, N., Iwamoto, M., Enomoto-Iwamoto, M., Noji, S., Ohuchi, H and Yoshioka, H., et al. 1997. Fibroblasts expressing sonic hedgehog induce osteoblast differentiation and ectopic bone formation. FEBS Letters 404, 319-323 https://doi.org/10.1016/S0014-5793(97)00014-8
  22. Lee, Y. S. 2001. Effect of isoflavones on proliferation and oxidative stress of MC3T3-E1 osteoblast-like cells. Korea Soybean Digest 18, 35-42
  23. Letton, R. W. 1990. Regulation of 25-hydroxyvitamin D3 metabolism in cultures of osteoblastic cells. J. Bone Miner. Res. 5(8), 815-820 https://doi.org/10.1002/jbmr.5650050804
  24. Maeda, T., Matsunuma, A., Kawane, T and Horiuchi, N. 2001. Simvastatin promotes osteoblast differentiation and mineralization in MC3T3-E1 cells. Biochem. Biophy. Res. Comm. 280, 874-877 https://doi.org/10.1006/bbrc.2000.4232
  25. Mori, Y., Hiraki, Y., Shukunami, C., Kakudo, S., Shiokawa, M and Kagoshima, M., et al. 1997. Stimulation of osteoblast proliferation by the cartilage-derived growth promoting factors chondromadulin-Ⅰand -Ⅱ. FEBS Letters 406, 310-314 https://doi.org/10.1016/S0014-5793(97)00291-3
  26. Orcel, P and Krane, S. M. 2000. Secondary osteoporosis and glucocorticoid induced osteoporosis. Ann. Med. Internal. 151, 497-502
  27. Owen, T. A., Aronow, M., Shalhoub, V., Barone, L. M., Wilming, L and Tassinari, M. S., et al. 1990. Progressive development of the rat osteoblast in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J Cell Physiol. 143, 420-430 https://doi.org/10.1002/jcp.1041430304
  28. Parfitt, A. M. 1994. Osteonal and hemi-osteonal remodeling: The spatial and temporal framework for signal traffic in adult human bone. J. Cell. Biochem. 55, 273-286 https://doi.org/10.1002/jcb.240550303
  29. Park, J. M and Joo, K. J. 1982. Stability of Anthocyanin Pigment from Juice of Raspberries. J. Korean Soc. Food Sci. Nutr 11, 67-74
  30. Pfeilschifter, J and Wolf, O. 1990. Chemotactic response of osteoblastic cells to Transforming growth factor B. J Bone. Miner. Res. 5, 815-823 https://doi.org/10.1002/jbmr.5650050804
  31. Quarles, L. D., Yohay, D. A., Lever, L. W., Caton, R and Wenstrup, R. J. 1992. Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner. Res. 7, 683-692 https://doi.org/10.1002/jbmr.5650070613
  32. Ryan, P. J., Evans, P., Gibson, T and Fogelman, I. 1992. Osteoporosis and chronic back pain: A study with single- photon emission computed tomography bone scintigraphy. J. Bone Miner. Res. 7, 1455-1460 https://doi.org/10.1002/jbmr.5650071213
  33. Schiller, P. C., D'ippolito, G., Balkan, W., Roos, B. A and Howard, G. A. 2001. Gap-junctional communication is required for the maturation process of osteoblastic cells in culture. Bone 28, 362-369 https://doi.org/10.1016/S8756-3282(00)00458-0
  34. Stein, G. S., Lian, J. B and Owen, T. A. 1990. Relationship of cell growth to the regulation for tissue-specific gene expression during osteoblast differentiation. FASEB J 4, 3111-3123
  35. Stein, G. S., Lian, J. B., Wijnen, A. J and Montechino, M. 1996. Transcriptional control of osteoblast growth and differentiation. Physiol. Rew 76, 593-629
  36. Torii, Y., Hitomi, K., Yamagishi, Y and Tsukagoshi, N. 1996. Demonstration of alkaline phosphatase participation in the mineralization of osteoblasts by antisense RNA approach. Cell Biol. Int. 20, 459-464 https://doi.org/10.1006/cbir.1996.0060
  37. Weiss, M. J., Ray, K., Fallon, M. D., Whyte, M. P., Fedde, K. N and Lafferty, M. A., et al. 1989. Analysis of liver/bone/kidney phosphatase mRNA, DNA and enzymatic activity in cultured skin fibroblasts from 14 unrelated patients with severe hypophosphatasia. Am. J. Hum. Genet 44, 686-694
  38. Yamashita, Y., Sato, M and Noguchi, T. 1987. Alkaline phosphatase in the periodontal ligament of the rabbit and macaque monkey. Arch. Oral. Biol. 32, 677-678 https://doi.org/10.1016/0003-9969(87)90044-6
  39. Yoo, M. C., Han, C. S., Jang, S. G and Han, I. K. 1989. The effects of the several hormones on the MC3T3 osteoblast cells -in vitro study. J. of Korean Orthop. Assoc 24, 879-888
  40. Yoon, I., Wee, J. H., Moon, J. H., Ahn, T. H and Park, K. H. 2003. Isolation and Identification of Quercetin with Antioxidative Activity from the Fruits of Rubus coreanum Miquel. J. Food Sci. 35, 499-502 https://doi.org/10.1111/j.1365-2621.1970.tb00968.x
  41. Yoon, S. R., Jeong, Y. J., Lee, G. D and Kwon, J. H. 2003. Changes in Phenolic Compounds Properties of Rubi Fructus Extract Depending on Extraction Conditions. J. Korean Soc. Food Sci. Nutr. 32, 338-345 https://doi.org/10.3746/jkfn.2003.32.3.338

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