DOI QR코드

DOI QR Code

MC3T3-E1 조골세포주와 RAW 264.7 파골세포주에서 길경을 함유한 한약재 추출물의 항골다공증 효과

Anti-osteoporotic Activity of Mixed Herbal Extract Involving Platycodon Grandiflorum Root in Osteoblastic MC3T3-E1 and Osteoclastic RAW 264.7 Cells

  • 정재인 (한림대학교 식의약품 효능평가 및 기능성소재개발센터) ;
  • 이현숙 (동서대학교 식품영양학과) ;
  • 김형준 (세명대학교 한의과대학) ;
  • 김용민 (세명대학교 화장품과학과) ;
  • 김수현 (구운식품 기업부설연구소) ;
  • 유동진 (구운식품 기업부설연구소) ;
  • 김은지 (한림대학교 식의약품 효능평가 및 기능성소재개발센터)
  • Jung, Jae-In (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Lee, Hyun-Sook (Dept. of Food Science & Nutrition, Dong-Seo University) ;
  • Kim, Hyung-Joon (Korean Medicine College, Se-Myung University) ;
  • Kim, Yong-Min (Dept. of Cosmetic Sciences, Se-Myung University) ;
  • Kim, Soo-Hyun (Research Institute, Guwoon Food) ;
  • Yoo, Dong-Jin (Research Institute, Guwoon Food) ;
  • Kim, Eun-Ji (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University)
  • 투고 : 2018.09.19
  • 심사 : 2018.11.26
  • 발행 : 2018.11.30

초록

목 적: 본 연구는 길경(Platycodon grandiflorum root)를 함유한 복합 한약재 추출물(ExMH-PGR)의 골다공증 예방 및 치료효과를 알아보기 위해 MC3T3-E1 조골세포주와 RAW 264.7 파골세포주를 이용하여 in vitro 수준에서 검증하였다. 방 법: 배양액에 다양한 농도의 ExMH-PGR를 첨가하여 MC3T3-E1 세포와 RAW 264.7 세포의 세포증식을 비교하였다. MC3T3-E1 세포에서 Alkaline phosphatase (ALP) 활성, 콜라겐 합성, 오스테올칼신 생성, 무기질 축적을 분석하였다. RAW 264.7 세포에서 Tartrate-resistant acid phosphatase (TRAP) 활성과 actin ring 형성 정도를 분석하였다. 결 과: ExMH-PGR는 $25{\mu}g/mL$ 농도까지 유의한 수준으로 ALP 활성, 콜라겐 합성, 그리고 오스테올칼신 형성을 증가시켰다. $50{\sim}200{\mu}g/mL$의 ExMH-PGR은 TRAP 활성과 actin ring 형성을 유의하게 억제했다. 결 론: ExMH-PGR은 조골세포의 활성을 촉진하고 파골세포의 활성을 억제하는 효과가 있어 골다공증 치료에 유용할 가능성이 있다.

Objectives: Osteoporosis is considered a serious human disease. We developed an extract of mixed herbs containing root of Platycodon grandiflorum (ExMH-PGR), which is expected to be effective in preventing or treating osteoporosis. The aim of this study was to investigate the anti-osteoporotic effect of ExMH-PGR in osteoblastic MC3T3-E1 cells and osteoclastic RAW 264.7 cells. Methods: To examine the anti-osteoporotic effect of ExMH-PGR, osteoblast and osteoclast differentiation were induced and cultured with various concentrations of ExMH-PGR. Alkaline phosphatase (ALP) activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells were analyzed. Tartrate-resistant acid phosphatase (TRAP) activity and the formation of actin ring in RAW 264.7 cells were analyzed. Results: ExMH-PGR at concentration up to $25{\mu}g/mL$ significantly increased ALP activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells. ExMH-PGR at 50 to $200{\mu}g/mL$ significantly inhibited TRAP activity and the formation of actin ring in RAW 264.7 cells. Conclusions: These results demonstrate that ExMH-PGR stimulates osteoblastic activities and inhibits osteoclastic activities in in vitro systems, suggesting that ExMH-PGR might be considered as an anti-osteoporotic candidate for treatment of osteoporosis disease.

키워드

참고문헌

  1. van den Bergh JP, van Geel TA, Geusens PP. Osteoporosis, frailty and fracture: implications for case finding and therapy. Nat Rev Rheumatol. 2012;8(3):163-72. https://doi.org/10.1038/nrrheum.2011.217
  2. He JB, Chen MH, Lin DK. New insights into the tonifying kidney-yin herbs and formulas for the treatment of osteoporosis. Arch Osteoporosis. 2017; 12(1):14. https://doi.org/10.1007/s11657-016-0301-4
  3. Rossouw JE, et al. Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results from the Women's health initiative randomized controlled trial. JAMA 2002;288(3):321-33. https://doi.org/10.1001/jama.288.3.321
  4. O'Ryan FS, Lo JC. Bisphosphonaterelated osteonecrosis of the jaw in patients with oral bisphosphonate exposure: clinical course and outcomes. J Oral and Maxillofac Surg 2012;70(8):1844-53. https://doi.org/10.1016/j.joms.2011.08.033
  5. Zhang L, et al. Platycodon grandiflorus - an ethnopharmacological, phytochemical and pharmacological review. J Ethnopharmacol. 2015;164:147-61. https://doi.org/10.1016/j.jep.2015.01.052
  6. Ryu CS, et al. Evaluation of the total oxidant scavenging capacity of saponins isolated from Platycodon grandiflorum. Food Chem. 2012;132(1):333-7. https://doi.org/10.1016/j.foodchem.2011.10.086
  7. Ashok BT, Ali R. The aging paradox: free radical theory of aging. Exp Gerontol. 1999;34(3):293-303. https://doi.org/10.1016/S0531-5565(99)00005-4
  8. Halliwell B. Oxidative stress and neurodegeneration: where are we now? J Neurochem. 2006;97(6):1634-58. https://doi.org/10.1111/j.1471-4159.2006.03907.x
  9. Zhao HL, et al. Hypocholesterolemic and anti-obesity effects of saponins from Platycodon grandiflorum in hamsters fed atherogenic diets. J Food Sci. 2008; 73(8):H195-200. https://doi.org/10.1111/j.1750-3841.2008.00915.x
  10. Wu J, et al. Cholesterol metabolism regulation and antioxidant effect of platycodin D on hyperlipidemic emulsionreduced rats. Afr J Pharm Pharmacol. 2011;5(22):2444-53.
  11. Choi YH, et al. Antiproliferative effects of saponins from the roots of Platycodon grandiflorum on cultured human tumor cells. J Nat Prod. 2010;73(11):1863-7. https://doi.org/10.1021/np100496p
  12. Zhang Z, et al. Platycodin D, a triterpenoid saponin from Platycodon grandiflorum, suppresses the growth and ivasion of human oral squamous cell carcinoma cells via the NF-kB pathway. J Biochem Mol Toxicol. 2017;31(9) e21934:1-7. https://doi.org/10.1002/jbt.21934
  13. Halliwell B. Oxidative stress and cancer: have we moved forward? Biochem J. 2007;401(1):1-11. https://doi.org/10.1042/BJ20061131
  14. Evans P, Halliwell B. Micronutrients: oxidant/antioxidant status. Br J Nutr. 2001;85(S2):S67-S74. https://doi.org/10.1079/BJN2000296
  15. Zhang L, et al. Anti-fungal and antibacterial activities of ethanol extracts of selected traditional Chinese medicinal herbs. Asian Pac J Trop Med. 2013; 6(9):673-81. https://doi.org/10.1016/S1995-7645(13)60117-0
  16. Ahn KS, et al. Platycodin D-induced apoptosis through nuclear factor-${\kappa}B$ activation in immortalized keratinocytes. Eur J Pharmacol. 2006;537(1-3):1-11. https://doi.org/10.1016/j.ejphar.2006.03.012
  17. Jeong HM, et al. Saponins from the roots of Platycodon grandiflorum stimulate osteoblast differentiation via p38 MAPK- and ERK-dependent RUNX2 activation. Food Chem Toxicol. 2010; 48(12):3362-8. https://doi.org/10.1016/j.fct.2010.09.005
  18. Choi JH, et al. Platycodin D inhibits osteoclastogenesis by repressing the NFATc1 and MAPK signaling pathway. J Cellular Biochem. 2017;118(4):860-8. https://doi.org/10.1002/jcb.25763
  19. Kim EJ, et al. Trans-10,cis-12-conjugated linoleic acid inhibits Caco-2 colon cancer cell growth. Am J Physiol Gastrointest Liver Physiol. 2001;283(2):357-67. https://doi.org/10.1152/ajpgi.00495.2001
  20. Puchtler H, Meloan SN, Terry MS. On the history and mechanism of Alizarin red S stains for calcium. J Histochem Cytochem. 1969;17(2):110-24. https://doi.org/10.1177/17.2.110
  21. Soya NS, Alles N. Osteoclast function and bone-resorbing activity: An overview. Biochem Biophys Res Commun. 2016; 476(3):115-20. https://doi.org/10.1016/j.bbrc.2016.05.019
  22. Scalbert A, et al. Dietary polyphenols and the prevention of diseases. Crit Rev Food Sci Nutr. 2005;45(4):287-306. https://doi.org/10.1080/1040869059096
  23. Hwang YP, et al. Saponins, especially platycodin D, from platycodon grandiflorum modulate hepatic lipogenesis in high-fat diet-fed rats and high glucose-exposed HepG2 cells. Toxicol Appl Pharmacol. 2013;267(2):174-83. https://doi.org/10.1016/j.taap.2013.01.001
  24. Ann JY, et al. A review of antiosteoporosis effects of herbal extracts in Korean journals. The Korean Journal of Oriental Medical Prescription. 2008;16(2):47-69.
  25. Kim M. Effect of Platycodon Grandiflorum A. extract in bone metabolism in ovariectomized rats. Korean J. Oriental Physiology & Pathology. 2008;22(1):183-8.
  26. Moon NY, Kim DC, Baek SH. Effect of Hyulbuchukeotang on inhibition of proliferation of uterine leiomyoma cells and cell apoptosis. The Journal of Oriental Obstetrics and Gynecology. 2006;19(2):186-98.
  27. Tang W, Eisenbrand G. Chinese drugs of plant origin. Heidelberg:Springer. 1992:113-25, 199-201, 539-43, 567-88, 703-10, 1011-5.
  28. Gou X, He G, Guo X. Poria cocos (Schw.) Wolf 茯苓(Fuling, Indian Bread). Dietary Chinese Herbs. 2015;87:781-8.
  29. Sharma U, Pal D, Prasad R. Alkaline phosphatase: an overview. Indian J Clin Biochem. 2014;29(3):269-78. https://doi.org/10.1007/s12291-013-0408-y
  30. Daneault A. Hydrolyzed collagen contributes to osteoblast differentiation in vitro and subsequent bone health in vivo. Osteoarthritis and Cartilage. 2014:22:S131.
  31. Lian JB, Stein GS. Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formation. Crit Rev Oral Biol Med. 1992;3(3):269-305. https://doi.org/10.1177/10454411920030030501
  32. Lee NK, et al. Endocrine regulation of energy metabolism by the skeleton. Cell. 2007;130(3):456-69. https://doi.org/10.1016/j.cell.2007.05.047
  33. Lee HW, et al. Berberine promotes osteoblast differentiation by Runx2 activation with p38 MAPK. J Bone Miner Res. 2008;23(8):1227-37. https://doi.org/10.1359/jbmr.080325
  34. Satue M, et al. Quercitrin and taxifolin stimulate osteoblast differentiation in MC3T3-E1 cells and inhibit osteoclastogenesis in RAW 264.7 cells. Biochem Pharmacol. 2013;86(10):1476-86. https://doi.org/10.1016/j.bcp.2013.09.009
  35. Zhou L, et al. Berberine sulfate attenuates osteoclast differentiation through RANKL induced NF-kappaB and NFAT pathways. Int J Mol Sci. 2015;16(11):27087-96. https://doi.org/10.3390/ijms161125998
  36. Teitelbaum SL. Bone resorption by osteoclasts. Science. 2000;289(5484):1504-8. https://doi.org/10.1126/science.289.5484.1504
  37. Hayman AR. Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy. Autoimmunity. 2008;41(3):218-23. https://doi.org/10.1080/08916930701694667
  38. Teitelbaum SL. The osteoclast and its unique cytoskeleton. Ann. N.Y. Acad. Sci. 2011;1240(1):14-7. https://doi.org/10.1111/j.1749-6632.2011.06283.x
  39. Schwayer C, et al. Actin rings of power. Dev Cell. 2016;37(6):493-506. https://doi.org/10.1016/j.devcel.2016.05.024
  40. Lee SK, et al. Platycodin D blocks breast cancer-induced bone destruction by inhibiting osteoclastogenesis and the growth of breast cancer cells. Cell Physiol Biochem. 2015;36(5):1809-20. https://doi.org/10.1159/000430152

피인용 문헌

  1. 골다공증의 한방 복합 처방 및 단일 본초에 대한 국내 연구 고찰 - 2013년 이후 발표된 국내 연구를 중심으로 vol.35, pp.3, 2021, https://doi.org/10.15188/kjopp.2021.06.35.3.87