Effects of Jeopgolsan (JGS) Extract on Fracture Healing

접골산(接骨散)이 골절치유에 미치는 영향

  • Received : 2017.12.26
  • Accepted : 2018.01.12
  • Published : 2018.01.31


Objectives The purpose of this study was to evaluate the effect of Jeopgolsan (JGS) extract on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells and on factors related with fracture healing in skull fractured rat. Methods Experimental animals were divided into four groups: normal group without any treatment (Normal), contral group were treated orally with distilled water (Control), Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 200) and Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 400). Rats in each group except the normal group were induced fractures in the skull. The 1,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity were measured to evaluate antioxidant activity. The production of nitric oxide (NO), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to evaluate anti-inflammatory activity. The production of osteocalcin calcitonin, carboxy-terminal telepeptides of type II collagen (CTX II), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2), Insulin and alkaline phosphatase (ALP) in serum of rats were measured to evaluate the effects of fracture healing at 0, 2, 4, and 6th week. X-rays were taken every 3 week from 0 to 6th week to evaluate fracture healing effect. Results 1. No cytotoxicity was observed. 2. DPPH and ABTS radical scavenging activity were increased in a concentration dependent manner, indicating anti-oxidant effect. 3. NO, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ were not significantly changed, indicating no anti-inflammatory effect. 4. Osteocalcin, Calcitonin, $TGF-{\beta}$ and ALP were significantly increased in the experimental groups. 5. CTX II, insulin were significantly decreased in the expermental groups. 6. Radiologic examination showed that union of fracture was promoted. Conclusions From above results, JGS showed significant results in factors related with fracture healing and radiologic examination. Threfore, JGS is expected to be effective in the treatment of fracture.


Jeopgolsan (JGS);Anti-oxidant;Anti-inflammation;Fracture


  1. Helvering LM, Sharp RL, Ou X, Geiser AG. Regulation of the promoters for the human bone morphogenetic protein 2 and 4 genes. Gene. 2000;256(1-2):123-38.
  2. Sandberg MM, Aro HT, Vuorio EI. Gene expression during bone repair. Clinical Orthopaedics and Related Research. 1993;289:292-312.
  3. Minina E, Wenzel HM, Kreschel C, Karp S, Gaffield W, McMahon AP, Vortkamp A. BMP and Ihh/PTHrP signaling interact to coordinate chondrocyte proliferation and differentiation. Development. 2001;128(22):4523-34.
  4. Kawai S, Sugigura T. Characterization of human bone morphogenetic protein(BMP)-4 and -7 gene promoters, activation BMP promotes by Gli, a sonic hedgehog mediator. Bone. 2001;29(1):54-61.
  5. Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahachi N, Ikeda T, Rosen V, Wozney JM, Fujisawa-Sehara A, Suda T. Bone morphogenetic protein-2 converts the diffenrentiation pathway of C2C12 myoblasts into the osteoblast lineage. Journal of Cell Biology. 1994;127:1755-66.
  6. Sampath TK, Muthukumaran N and Reddi AH. Isolation of osteogenin, an extracellular matrix associated, bone inductive protein, by heparin affinity chromatography. Proceedings of the National Academy of Sciences. 1987;84:7109-13.
  7. Urist MR, DeLange RJ, Finerman GAM. Bone cell differentiation and growth factors. Science. 1983;220:680-6.
  8. Lee KH, Lee YI, Cho KC, Oh IS, Lee JY, Kim SJ. Expression of TGF-${\beta}$ I and II ligands and receptors at epiphyseal plate and fracture callus. Journal of Korean Orthopedic. 1998;33(2):458-65.
  9. Nanes MS, Rubin J, Titus L, Hendy GN, Catherwood B. Tumor necrosis factor-alpha inhibits 1,25-dihydroxyvitamin D3-stimulated bone Gla protein synthesis in rat osteosarcoma cells (ROS 17/2.8) by a pretranslational mechanism. Endocrinology. 1991;128(5):2577-82.
  10. Felix R, Fleisch H, Elford PR. Bone-resorbing cytokines enhance release of macrophage colony-stimulating activity by the osteoblastic cell MC3T3-E1. Calcified Tissue International. 1989;44(5):356-60.
  11. Pfeilschifter J, Chenu C, Bird A, Mundy GR, Roodman GD. Interleukin-1 and tumor necrosis factor stimulate the formation of human osteoclastlike cells in vitro. Journal of Bone and Mineral Research. 1989;4(1):113-8.
  12. Thomson BM, Saklatvala J, Chambers TJ. Osteoblasts mediate interleukin 1 stimulation of bone resorption by rat osteoclasts. Journal of Experimental Medicine. 1986; 164(1):104-12.
  13. Ishimi Y, Miyaura C, Jin CH, Akatsu T, Abe E, Nakamura Y, Yamaguchi A, Yoshiki S, Matsuda T, Hirano T. IL-6 is produced by osteoblasts and induces bone resorption. Journal of Immunology. 1990;145(10): 3297-303.
  14. Song KY. Seoul:Korea medical book publishing company. 1998:772.
  15. Seong SC, Choi MH, Lee MC Park SC, Song KY. Expression of osteocalcin and transglutaminase C during fracture healing and distraction osteogenesis in rat's tibia. The Journal of the Korean Orthopaedic Association. 1994;29(5):1312.
  16. Kim DY. Biochemical markers of bone turnover. Korean Journal Nuclear Medicine. 1999;33(4):341-51.
  17. Ross MH, Romred LJ, Kaye GI. Histology a text and atlas 3rd edition. Williams and Wilkins publisher. 1995: 150-87.
  18. Lee MS, Kim SY, Lee MC, Cho BY, Lee HK, Koh CS, Min HK. Negative correlation between the change in bone mineral density and serum osteocalcin in patients with hyperthyroidism, The Journal of Clinical Endocrinology and Metabolism. 1990;70:766-70.
  19. Song JH, Han KO, Kim DY, Woo JT, Kim SW, Yang IM, Kim JW, Kim YS, Choi YK. Markers of bone turnover and bone mineral density in hyperthyroidism and effects of treatment. The Korean Society of Bone Metabolism. 1994;1:177-85.
  20. Hauschka PV, Carr SA. Calcium-dependent ahelical structure in osteocalcin. Biochemistry. 1982;21:2538-47.
  21. Price PA, Parthermore JG, Doftos LJ. New biochemical marker for bone metabolism. Journal of Clinical Investigation. 1980;66:878-83.
  22. Kim SW. Clinical endocrinology of calcitonin. Journal of Korean Society of Endocrinology. 1991;6(2):117.
  23. Johason JS, Riis BJ, DElmas PD. Plasama BGP. An indicator of spontaneous bone loss and of effect of estrogen treatment in postmenopausal women. European Journal of Clinical Investigation. 1998;18:191-5.
  24. Joyce ME, Robert AB, Sporn MB, Bolander ME. Transforming growth factor-${\beta}$ in the regulation of fracture repair. The Orthopedic Clinics of North America. 1990; 21(1):199-209.
  25. Bondet V, Band-Williams W, Berset C. Kinetics and mechanism of antioxidant activity using the DPPH free radical method. LWT-Food Science and Technology. 1997;30:609-15.
  26. Fellegrini N, Ke R, Yang M, Rice-Evans C. Screening of dietary carotenoids and carotenoid-rich fruit extracts for antioxidant activities applying 2,2'-azinobis (3-ethylenebenzothiazoline- 6-sulfonic acid) radical cation decolorization assay. Methods in Enzymology. 1999;299:379-89.
  27. Dee Unglaub Silverthorn. Human physiology 6th edition. Seoul:Life Science. 2014:820.
  28. Vinay Kumar, Abul K. Abbas, Nelson Fausto. Pathologic basis of disease. Philadelphia:Elsevier. 2005:47-86.
  29. Storck M, Schilling M, Prestel R, Abendroth D, Burkhardt K, Prestel R, Hammer C. Production of proinflammatory cytokines and adhesion molecules in ex-vivo xenogeneic kidney perfusion. Transplant International. 1994;7(1):647-9.
  30. Moncada S, Palmer RM, Higgs EA. Nitric oxide:physiology, pathology and pharmacology. Pharmacological Reviews. 1991;43:109-42.
  31. David A. Winka, James B. Mitchell. Chemical biology of nitric oxide: insights into regulatory, cytotoxic, and cytoprotective mechanisms of nitric oxide. Free Radical Biology and Medicine. 1998;25(4-5):434-56.
  32. Kawamata H, Ochiai H, Mantani N, Terasawa K. Enhanced expression of inducible nitric oxide synthase byJuzen-taiho-to in LPS-activated RAW264.7 cells, a murine macrophage cell line. Americal Journal of Chinese Medicine. 2000;28(2):217-26.
  33. Lee BG, Kim SH, Zee OP, Lee KR, Lee HY, Han JW, Lee HW. Suppression of inducible nitric oxide synthase expression in RAW 264.7 macrophages by two-carboline alkaloids extracted from Melia azedarach. European Journal of Pharmacology. 2000;406(3):301-9.
  34. Seo WG, Pae HO, Oh GS, Chai KY, Yun YG, Kwon TO, Chung HT. Inhibitory effect of ethyl acetate fraction from Cudrania tricuspidata on the expression of nitric oxide synthase gene in RAW 264.7 macrophages stimulated with interferon-and lipopolysaccharide. General Pharmacology. 2000;35(1):21-8.
  35. Chiou WF, Chou CJ, Chen CF. Camptothecin suppresses nitric oxide biosynthesis in RAW 264.7 macrophages. Life Science. 2001;69:625-35.
  36. Seo WG, Pae HO, Oh GS, Kim NY, Kwon TO, Shin MK, Chai KY, Chung HT. The aqueous extract of Rhodiola sachalinensis root enhances the expression of inducible nitric oxide synthase gene in RAW264.7 macrophages. Journal of Ethnopharmacolgy. 2001;76(1):119-23.
  37. Kim CH, Kim DK, Park SI, Sohn KH, Kim GS. Effects of interleukin-6 on mRNA expression of alkaline phospahatase, osteocalcin, decorin and ${\alpha}$1(1)-collagen in human bone marrow stromal cell. Endocrinology and Metabolism. 1996;11(2):156-62.
  38. Centrella M, McCarthy TL, Canalis E. Tumor necrosis factor- alpha inhibits collagen synthesis and alkaline phosphatase activity independently of its effect on deoxyribonucleic acid synthesis in osteoblast-enriched bone cell cultures. Endocrinology. 1988;123(3):1442-8.
  39. Yim CB, Kim YJ, Oh MS. The oriental and western medical study of fracture. Daejeon University, Institute of Korean Medicine. 2007;16(1):157-66.
  40. The Korean Orthopaedic Association. Orthopaedics the 7th edition. Seoul:New medical publisher. 2013:87, 94-5, 1161-2, 1385-94.
  41. Yeom IH, Oh MS, Song TW. Helling effect of Gamigungguitang and Gamigungguit-angGaNokyong water extract on tibia fractured rats. Daejeon University, Institute of Kore-an Medicine. 1999;15:683.
  42. The Korean Fracture Society. Principles of Fracture Management. Seoul:Panmuneducation. 2013:3,10,40.
  43. The Society of Korean Medicine Rehabilitation. Korean medicine rehabilitation. 2nd edition. Seoul:Koonja publisher 2008:201.
  44. Bae KJ, Jeong JW, Jung MY, Kim SJ. Reviewing research on the treatment and study of fracture in Korean journals objective-Focus on domestic thesis. Journal of Korean Medicine Rahabilitation. 2015;25(3):27-8.
  45. Heo Joon. Donguibogam. Seoul:Hanmi publisher. 2001: 934-5.
  46. Laufer S, Greim C, Bertsche T. An in-vitro screening assay for the detection of inhibitors of proinflammatory cytokine synthesis: a useful tool for the development of new antiarthritic and disease modifying drugs. Osteoarthritis and Cartilage. 2002;10:960-7.
  47. John W, Hole J. Human anatomy physiology Wm. C Brown publisher Oxford. 1993:170-227.
  48. Hulth A. Current concepts of fracture healing. Clinical Orthopaedics & Related Research. 1989;249:265-84.
  49. Forst HM. The biology of fracture healing: An overview for clinicians. Part I. Clinical Orthopaedics & Related Research. 1989;248:283-93.
  50. Korean health industry development institute. Survey of utilization of Korean medical services and consumption of herbal medicine. 2014:129.
  51. Korean medical herbalogy professors. Herbalogy. Seoul:Younglimsa. 2000:316, 411-2, 436, 492, 614.
  52. Kim SH, Choi HJ, Oh HT, Cheng BC, Ham SS. Cytoprotective effect by antioxidant activity of Codonopsis lanceolata and Platycodon grandiflorum ethyl acetate fraction in human HepG2 cells. Korean Journal of Food Science and Technology. 2008;40(6):696-701.
  53. Videla LA, Fermandez V. Biochemical aspects of cellular oxidative stress. Archivos de Biologia y Medicina Experimentales. 1998;21(1):85-92.
  54. Kondo T, Hirose M, Kageyama K. Roles of oxidative stress and redox regulation in atherosclerosis. Journal of Atherosclerosis and Thrombosis. 2009;16(5):532-8.
  55. Fridovich I. Superoxide dismutases. An adaptation to a paramagnetic gas. The Journal of Biological Chemistry. 1989;264(14)7761-4.
  56. Shon MS, Song JH, Kim JS, Jang HD, Kim Gn. Antioxidant activity of oil extracted from Korean red Ginseng and its moisturizing function. Korean Journal of Aesthet Cosmetol. 2013;11(3):489-94.
  57. Sheweita SA, Khoshhal KI. Calcium metabolism and oxidative stress in bone fractures: Role of antioxidants. Current Drug Metabolism. 2007;8(5):519-25.
  58. Health Insurance Review & Assessment Service. Fracture patient statistics 2011 Jan-2015 Dec. [cited 2016 Nov 22]. Available from:URL:
  59. Paglia D, Wey A, Breitbart EA, Faiwiszewski J, Mehta SK, Al-Zube L, Vaidya S, Cottrell JA, Graves D, Benevenia J, O'Connor JP, Lin S. Effects of local insulin delivery on subperiosteal angiogenesis and mineralized tissue formation during fracture healing. Journal of Orthopaedic Research. 2013;31:783-91.
  60. Kim DY. Biochemical markers of bone formation. Korean Journal of Bone Metabolism. 1994;1(1):233-9.
  61. Yoo J, Johnstone B. The role of osteochondral progenitor cells in fracture repair. Clinical Orthopaedics and Related Research. 1998;335:73-81.
  62. Lee HS, Lee CS, Jang JS, Lee JD, Um SM. Changes of serum alkaline phophatase and osteocalcin during fracture healing. The Journal of the Korean Orthopaedic Association. 2002;37(3):411-5.