Journal of Korean Medicine Rehabilitation (한방재활의학과학회지)

The society of Korean Medicine Rehabilitation (한방재활의학과학회)
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Healing Effect of Danggwisu-san (Dangguixu-san) on Femur Fractured Mice

당귀수산(當歸鬚散)이 대퇴골절 유발 생쥐에 미치는 영향

  • Jeon, Dong-Hwi (Department of Korean Medicine Rehabilitation, College of Korean Medicine, Daejeon University) ;
  • Oh, Min-Seok (Department of Korean Medicine Rehabilitation, College of Korean Medicine, Daejeon University)
  • 전동휘 (대전대학교 한의과대학 한방재활의학교실) ;
  • 오민석 (대전대학교 한의과대학 한방재활의학교실)
  • Received : 2020.12.14
  • Accepted : 2021.01.07
  • Published : 2021.01.31
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Abstract

Objectives This study was designed to evaluate the effects of Danggwisu-san (Dangguixu-san, DG) on bone repair from femur fracture in mice. Methods Mice were randomly divided into 4 groups (normal, control, positive control and DG 300 mg/kg-treated group). In order to investigate the effects of DG on gene expressions in experimental animals with fracture, we measured the levels of bone morphogenetic protein-2 (BMP2), cyclooxygenase-2 (COX2), Sox9, collagen type II alpha 1 chain (Col2a1), runt-related transcription factor 2 (Runx2), osterix genes. After the cytotoxicity test, we analyzed the levels of expression of osteocalcin and Runx2, and tumor necrosis factor-α (TNF-α), a pro-inflammatory cytokine. The process of fusion in the fracture was also investigated by gross examination. Results Through in vivo BMP2, COX2 gene expression significantly decreased. Sox9 significantly increased. Col2a1, Runx2, osterix gene expression also increased as well, but there was no statistical significance. The degree of unilateral fracture fusion investigated by gross examination was significantly faster than those of the other groups. Through in vitro the level of TNF-α in macrophages was increased by DG in a dose-dependent mannerand and 250 and 500 ㎍/mL showed statistical significance. Osteocalcin and Runx2 genes expressions increased when DG was treated in osteoblasts. Conclusions DG promotes the healing of the fracture through the expression of bone repair-related genes and TNF-α production. This study may set the foundation for the clinical application of DG to the patients with bone fractures.

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References

  1. The Society of Korean Medicine Rehabilitation. Korean Medicine Rehabilitation. 4th ed. Paju:Koonja Publishing. 2015:206-10.
  2. The Korean Orthopaedic Association. Orthopaedics. 6th ed. Seoul:ChoiSin medical Publishing Co. 2006:779-831.
  3. Shin KM, Jung CY, Hwang MS. Effects of administration of pyritum on fracture healing in mice. The Journal of Korean Acupuncture & Moxibustion Society. 2009;26(5):65-75.
  4. Lee HK, Chung MS, Gang SY. Basic study on the effect of Korean Ginseng upon fracture healing of the bone. The Journal of the Korean Orthopaedic Association. 1984;19(3):483-91. https://doi.org/10.4055/jkoa.1984.19.3.483
  5. Suk SI. Orthopedics. Seoul:The New Medical Journal. 2013:1165-75.
  6. Kim JH. The effects of Korean safflower seed powder supplementation diet on the recovery of rib fracture and the improvement lipid metabolism in rats, and development of processed food [dissertation]. Daegu: Kyoungbuk University; 1998:1-7.
  7. Jeon SM, Kim JH, Lee HJ. The effects of Korean Safflower (Carthamus tinctorious L.) seed powder supplementation diet on bone metabolism indices in rats during the recovery of rib fracture. Journal of Nutrition and Health. 1998;31(6):1049-56.
  8. Geum DH, Kim SS. Healing effect of pyrite on tibia fractured rats. J Korean Med Rehabil. 2002;12(2):51-68.
  9. Hang TG, Oh MS, Song TW. Helling effect of Sintongchugoetang water extract on tibia fractured rats. Daejeon University, Institute of Korean Medicine. 1999;8(1):727-38.
  10. Pyum YH, Oh MS, Song TW. Helling effect of Gamigungguitang and GamigungguitangGaNokyong water extract on tibia fractured rats. Daejeon University Institute of Korean Medicine. 1999;8:675-87.
  11. Son WT, Song TW, Oh MS. Healing effect of Soongiwhalhyultang extract on tibia fractured rats. J Korean Med Rehabil. 1999;9(2):350-62.
  12. Ahn HL, Shin MS, Kim SJ. Effects of Neutral Eohyeol (Yuxue) herbal acupuncture and DangkisoosanDangkisoosan (Dangguixu-san) on fracture healing in the early stage in rats. J Korean Med Rehabil. 2007;17(1):1-16.
  13. Kum DH, Kim SS. Healing effect of Bokwonhwalhyultang on tibia fractured. The Journal of the Korea Institute of Oriental Medical Informatics. 2002;8(1):47-66.
  14. Park BC, Cha YY, Lee ES. Study on the polyphosphate content of the Yukmijihwang-tang and its effect on transcription activity of genes related to bone morphogenesis. Korean Journal of Oriental Physiology & Pathology. 2004;18(6):1769-76.
  15. Shin YW, Park YI, Kim HY. Effect of Yukmigihwang-tang kamibang on the expression of osteo-related genes, TG2 and BMP4. The Journals of the Korean Oriental Medical Society. 2002;23(2):190-7.
  16. Kim GH. Healing effect of Gamiyungmijihwangtang on fractured dogs [dissertation]. Iksan:Wonkwang University; 1987:1-15.
  17. Kee YB, Kim DH, Kang DH. Effects of Yukmijihwang-tang (Liuweidihuang-tang) and cervi pantotrichum cornu pharmacopuncture on fracture healing in diabetic rats. Journal of Oriental Rehabilitation Medicine. 2012;22(3):49-63.
  18. Han SW, Choi JY, Lee YH. Healing of bony defects by cervi pantotricuhum cornu herbal acupuncture. Journal of Korean Acupuncture & Moxibustion Medicine Society. 2001;18(5):135-46.
  19. Lee C. Uihakipmun. Seoul:Namsandang. 1991:2201.
  20. Heo J. Donguibogam. Seoul:Yeo Gang Publishing House. 2002:2307-8.
  21. Jung IM. Effects of Dangkwisoo-san and native copper on TGF-β1 expression in fractured rats [dissertation]. Naju:Dongshin University; 2017:1-14.
  22. Bonnarens F, Einhorn TA. Production of a standard closed fracture in laboratory animal bone. J Orthop Res. 1984;2:97-101. https://doi.org/10.1002/jor.1100020115
  23. Michael H, Regis J. The convergence of fracture repair and stem cells: interplay of genes, aging, environmental factors and disease. Journal of Bone and Mineral Research. 2014;11(29):2307-22.
  24. Bae WS. The new oriental medical clinician. Seoul: Uiwondang. 1977:113-4.
  25. Wang JS. Question and answer of Chinese medicine basic theory. Hyupseo:Hyupseo Scientific Technique publishing. 1981:115-7.
  26. Shanghai Chinese Medical School. The foundation of Chinese medicine. Hongkong:Sangmuinseoguan. 1979:39, 43, 116.
  27. Yoon GY. A study on the methodology of oriental medicine. Seoul:Sungbosa. 1983:103.
  28. Yeo NH, Lee HY. The effect of Dangkwisoo-San on blood enzyme activity in brusied rats. The Korean Journal of Physical Education. 2002;41(3):333-40.
  29. Bak JW, Sim BY, Kim DH. The effects of Danggwisusan on restoration ability in wound induced animal models. The Korea Journal of Herbology. 2014;29(5):55-63. https://doi.org/10.6116/kjh.2014.29.5.55
  30. Yoon YG. Dongui antisepticology and prescription description. Seoul:Uiseongdang. 2011:679-80.
  31. Kim HJ, Oh WM, Koh JT. Effects of angiopoietin-1 on bone morphogenetic protein-2 induced osteoblast differentiation. Jeonnam Dental Collection of Dissertations. 2007;19(1):182-99.
  32. Heldin CH, Miyazono K, Dijke PT. TGF-β signaling from cell membrane to nucleus through SMAD proteins. Nature. 1997;390:465-71. https://doi.org/10.1038/37284
  33. Groszmann RJ. Hyperdynamic state in chronic liver diseases. J Hepatol. 1993;17(2):S38-40. https://doi.org/10.1016/S0168-8278(05)80454-4
  34. Rothkotter HJ, Pabst R, Bailey M. Lymphocyte migration in the intestinal mucosa: entry, transit and emigration of lymphoid cells and the influence of antigen. Vetrinary Immuno Immunopath. 1999;72:157-65. https://doi.org/10.1016/S0165-2427(99)00128-2
  35. Tsujii M, Kawano S, Tsuji S. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 1998;93(5):705-16. https://doi.org/10.1016/S0092-8674(00)81433-6
  36. Andrew BS, Yuehuei HA. Is there an inhibitory effect of COX-2 inhibitor on bone healing. Pharmacological Research. 2004;50(2):151-6. https://doi.org/10.1016/j.phrs.2003.12.017
  37. Christian C. General principles of fracture treatment. In: Canale ST ed. Camphell's operative orthopaedics. St. Louis:Mobsby. 1988:1993-2041.
  38. Brian HM, Spencer TC, Paul SW. Effect of COX-2 inhibitors and non-steroidal anti-inflammatory drugs on a mouse fracture model. Injury. 2006;37(9):827-37. https://doi.org/10.1016/j.injury.2005.12.018
  39. Spranger J. Bone dysplasia 'families'. Pathology and Immunopathology Research. 1988;7(1-2):76-80. https://doi.org/10.1159/000157098
  40. Cole WG. Collagen genes: mutations affecting collagen structure and expression. Progress in Nucleic Acid Research and Molecular Biology. 1994;47:29-80. https://doi.org/10.1016/S0079-6603(08)60249-4
  41. Ng LJ, Wheatly S, Muscat G. SOX9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse. Developmental Biology. 1996;138(1):108-21.
  42. Na SH. Localization of runx2 and osterix in the developing periodontal tissues of mouse [dissertation]. Seoul: Kyung Hee University; 2010:1-2.
  43. Nakajima F, Ogasawara A, Goto K, Moriya H. Spatial and temporal gene expression in chondrogenesis during fracture healing and the effects of basic fibroblast growth factor. Journal of Orthopaedic Research. 2001;19(5): 935-44. https://doi.org/10.1016/S0736-0266(01)00024-9
  44. Kim DY. Biochemical markers of bone turnover. Korean Society of Nuclear Meidicine. 1999;33(2):341-51.
  45. Price PA, Parthermore JG, Doftos LJ. New biochemical marker for bone metabolism. J Clin Invest. 1980;66:878-83. https://doi.org/10.1172/JCI109954
  46. Calvo MS, Eyre DR, Gundberg CM. Molecular basis and clinical application of biological markers of bone turnover. Endocr Rev. 1996;17:333-68. https://doi.org/10.1210/edrv-17-4-333
  47. Kim HS. Effect of dexamethasone and 1,25(OH)2D3 on proliferation and osteogenic differentiation of cultured human bone marrow stromal cells. Endocrinology and Metabolism. 2002;17(2):206-17.
  48. Beeton CA, Charfield D, Brooks RA. Circulating levels of interleukin-6 and its soluble receptor in patients with head injury and fracture. J Bone Joint Surg Br. 2004; 86:912-7.
  49. Einhorn TA, Majeska RJ, Rush EB, Levine PM, Horowitz MC. The expression of cytokine acitivity by fracture callus. J Bone Miner Res. 1995;10:1272-81. https://doi.org/10.1002/jbmr.5650100818
  50. Kimbke RB, Bain S, Pacifici R. The functional block of TNF but not IL-6 prevents bone loss in ovariectomized mice. J Bone Miner Res. 1997;12:935-41. https://doi.org/10.1359/jbmr.1997.12.6.935
  51. Kon T, Cho TJ, Aizawa T. Expression of osteoprotegerin, receptor activator of NF-kappaB ligand (osteoprotegerin ligand) and related proinflammatory cytokines during fracture healing. J Bone Miner Res. 2001;16:1004-14. https://doi.org/10.1359/jbmr.2001.16.6.1004