The effects of indomethacin on distribution and expression of COX-2 and IGF-I in the mandibular condyle of growing dogs

인도메타신투여가 개의 하악두에서 COX-2와 IGF-I의 발현과 분포에 미치는 영향

  • Nam, Jong-Hyun (Department of Dentistry, School of Dentistry, Kyung Hee University) ;
  • Lee, Ki-Soo (Department of Dentistry, School of Dentistry, Kyung Hee University) ;
  • Kang, Yoon-Goo (Department of Dentistry, School of Dentistry, Kyung Hee University)
  • 남종현 (경희대학교 치과대학 교정학교실) ;
  • 이기수 (경희대학교 치과대학 교정학교실) ;
  • 강윤구 (경희대학교 치과대학 교정학교실)
  • Published : 2005.10.01

Abstract

This study aimed to investigate the effects of indomethacin on distribution and expression of COX-2 and IGF-1 in the mandibular condyle ofi growing dogs and to examine the number of chondroclasts around the mineralization zone indomethacin inhibits prostatlandin $E_2$ production in the tissue by inhibiting synthesis of cyclooxygenase 2. Prostaglandin $E_2$ stimulates insulin-like growth factor synthesis. Insulin-like growth factor stimulates growth of mandibular condylar cartilage. Eight mongrel dogs. aged 13-14 weeks, were divided into 4 groups. Group 1 and group 2 were administered indomethacin 2 mg/Kg/day orally two times a day for 7 days and 14 days respectively. Group 3 were administered indomethacin 8mg/Kg/day orally 2 times a day for 14 days, and 4he control group were administered a placebo. The mandibular condyle heads were sectioned in $5{\mu}m$ thickness The specimens were stained with H-E staining. COX-2 immunohistochemical staining and IGF-1 immunohistochemical staining and examined under microscope. After TRAP staining, the number of chondroclasts were calculated The observed results were as follows: Indomethacin inhibited expression and distribution of COX-2 and IGF-1 on the proliferative zone of condylar cartillage. Indomethacin decreased the number of chondroclastes on the mineralization zone by a time-dependent manner (P<0.05). Indomethacin inhibited expression and distribution of IGF-I by a dose and time-dependent manner. These results show that indomethacin inhibited expression and distribution of COX-2 and IGF-1 on the proliferative zone of condylar cartilage and decreased the number of chondroclasts and suggests that when indomethacin is administered for a long time, condyle growth could be delayed.

이 연구는 $PGE_2$ 생합성 억제제인 인도메타신의 투여 시 성장기 개의 하악두 연골에 나타나는 cyclooxygenase-2 (GOX-2)와 insulin-like growth factor 1(IGF-1)의 발현과 분포를 관찰하여 인도메타신이 하악두 연골 성장에 미치는 영향을 규명하기 위하여 시행하였다 생후 12- 3주된 잡견 8마리를 4군으로 구분하였다 통상적 복용량인 인도메타신 2mg/Kg/day을 각각 7일과 14일간 투여한 군 과량인 8mg/Kg/day을 14일간 투여한 군과 무처치군인 대조군으로 구분하였으며. 하악두를 연구대상으로 하였다. 연구대상 하악두는 $5{\mu}m$ 두께의 절편을 만들어, H-E 중염색, COX-2 면역염색 IGF-1 면역염색을 시행하여 광학 현미경으로 검경하였으며, tartrate resistant acid phosphatase(TRAP) 염색 후 파연골세포의 수를 측정하여 다음과 같은 결과를 얻었다. 인도메타신은 하악두 연골의 증식대에서 COX-2와 IGF-1의 발현과 분포를 억제시켰으며, 인도메타신의 투여기간에 비례해서 파연골세포 수는 유의성 있게 감소하였다 (p<0.01) IGF-1의 발현과 분포는 인도메타신의 투여 양과 기간에 비례하여 억제되었다 이상의 결과에 의하면 인도메타신의 투여는 하악두 연골에서 COX-2와 IGF-1의 발현과 분포를 억제하고 파연골세포의 수를 감소시켜 하악두 성장을 억제할 가능성이 있음을 시사한다.

Keywords

References

  1. Copray JC, Jansen HW, Duterloo HS. Growth and growth pressure of mandibular condylar and some primary cartilages of the rat in vitro. Am J Ortbod Dentofacial Orthop 1986;90:19-28 https://doi.org/10.1016/0889-5406(86)90023-5
  2. Mizoguchi I, Takahashi I, Nakamura M, Sasano Y, Sato S, Kagayama M, et al. An immunohistochemical study of regional differences in the distribution of type I and type II collagens in rat mandibular condylar cartilage. Arch Oral BioI 1996:41:863-9 https://doi.org/10.1016/S0003-9969(96)00021-0
  3. Delatte M, Von den hoff JW, Maltha JC. Growth stimulation of mandibular condyles and femoral heads of newborn rats by IGF-1. Arch Oral Biol 2004:49:165-75 https://doi.org/10.1016/j.archoralbio.2003.09.007
  4. Lerner U. Indomethacin inhibits bone resorption in vitro without affecting bone collagen synthesis. Agents Actions 1982; 12:466-70 https://doi.org/10.1007/BF01965928
  5. O' Keefe RJ, Crabb ID, Puzas JE, Rosier RN. Influence of prostaglandins on DNA and matrix synthesis in growth plate chondrocytes. J Bone Miner Res 1992;7:397-404 https://doi.org/10.1002/jbmr.5650070407
  6. Klein DC, Raisz LG. Prostaglandin: stimulation of bone resorption in tissue culture. Endocrinology 1970;86: 1436-40 https://doi.org/10.1210/endo-86-6-1436
  7. Santoro MG, Jaffe BM, Simmons DJ, Bone resorption in vitro and in vivo in PGE-treated mice. Proc Soc Exp Biol Med 1977;156:373-7 https://doi.org/10.3181/00379727-156-39939
  8. Park YG, Chung KR, Lee YJ, Noh SH. Prostaglandin $E_2$ regulates the gene expression of bone matrix metalloproteinase-I in mouse osteoblasts. 경희치대논문집 2000;22:49-60
  9. Schwartz Z, Gilley RM, Sylvia VL., Dean DD, Boyan BD. The effect of prostaglandin $E_2$ on costochondral chondrocyte differentiation is mediated by cAMP and protein kinase C. Endocrinology 1998;139: 1825-34 https://doi.org/10.1210/en.139.4.1825
  10. Sylvia VL, Del Toro F Jr, Hardin RR, Dean DD, Boyan BD, Schwartz Z. Characterization of $PGE_2$ receptors (EP) and their role as mediators of 1, $25-(OH)_2D_3$ effects on growth zone chondrocytes. J Steroid Biochem Mol Biol 2001;78:261-74 https://doi.org/10.1016/S0960-0760(01)00099-1
  11. Suzuki S, Itoh K, Ohyama K. Local administration of IGF-I stimulates the growth of mandibular condyle in mature rats. J Orthod 2004;31: 138-43 https://doi.org/10.1179/146531204225020436
  12. Maor G, Laron Z, Eshet R, Silbermann M. The early postnatal development of the murine mandibular condyle is regulated by endogenous insulin-like growth factor-I. J Endocrinol 1993;137:21-6 https://doi.org/10.1677/joe.0.1370021
  13. Peacey SR. Shalet SM. Insulin-like growth factor I measurement in diagnosis and management of acromegaly. Ann Clin Biochem 2001;38: 297-303 https://doi.org/10.1258/0004563011900678
  14. DiBattista JA, Dare S, Morin N, Abribat T. Prostaglandin $E_2$ upregulates insulin-like growth factor binding protein- 3 expression and synthesis in human articular chondrocytes by a c- AMP-independent pathway: Role of calcium and protein kinase A and C. J Cell Biochem 1996:63:320- 33 https://doi.org/10.1002/(SICI)1097-4644(19961201)63:3<320::AID-JCB7>3.0.CO;2-Z
  15. Blanco FJ, Guitian R, Moreno J, de Toro FJ, Galdo F. Effect of antiinflammatory drugs on COX-1 and COX-2 activity in human articular chondrocytes. J Rheumatol 1999:26:1366-73
  16. Visnapuu V, Peltomaki T, Ronning O, Syrjanen S. Distribution of insulin-like growth factor-1 mRNA in the mandibular condyle and rib cartilage of the rat during growth. Arch Oral Biol 2002;47:791-8 https://doi.org/10.1016/S0003-9969(02)00115-2
  17. Miyamoto M, Mukai S, Kobayashi T, Yamamoto H, Kobayashi M, Maruyama T, et al. Simultaneous stimulation of EP2 and EP4 is essential to the effect of prostaglandin $E_2in$ chondrocyte differentiation. Osteoarthritis Cartilage 2003:11:644-52 https://doi.org/10.1016/S1063-4584(03)00118-3
  18. Banovac K, Williams JM, Patrick LD, Haniff YM. Prevention of heterotopic ossification after spinal cord injury with indomethacin. Spinal Cord 2001;39:370-4 https://doi.org/10.1038/sj.sc.3101166
  19. Maor G, Hochberg Z, Silbermann M. Insulin-like growth factor I accelerates proliferation and differentiation of cartilage progenitor cells in cultures of neonatal mandibular condyles. Acta Endocrinol 1993:128:56-64
  20. Lin WW, Oberauer AM. Spatiotempcral expression of alternatively spliced IGF - I mRNA in the rat costochondral growth plate. J Endocrinol 1999:160:461-7 https://doi.org/10.1677/joe.0.1600461
  21. Jee WS, Ueno K, Deng YP, Woodbury DM. The effects of prostaglandin $E_2$ in growing rats: Increased metaphyseal hard tissue and cortico-endostcal bone formation. Calcif Tissue Int 1985:37:148-57 https://doi.org/10.1007/BF02554834
  22. Hakada Y, Yoshino T, Nakatani Y, Kurihara N, Maeda N, Kumegawa M. Prostaglandin $E_2$ stimulates DNA synthesis by a cAMP-independent pathway in osteoblastic clone MC3T3-E1 cells. J Cell Physiol 1986:128:155-61 https://doi.org/10.1002/jcp.1041280204
  23. Hock JM, Centrella M, Canalis E. Insulin-like growth factor I has independent effects on bone matrix formation and cell replication. Endocrinology 1988:122:254-60 https://doi.org/10.1210/endo-122-1-254
  24. McCarthy TL, Centrella M, Raisz LG, Canalis E. Prostaglandin $E_2$ stimulates insulin-like growth tactor-1 synthesis in osteoblatenriched cultures from fetal rat bone. Endocrinology 1991;128:2895-900 https://doi.org/10.1210/endo-128-6-2895
  25. Soekanto A. Inhibition of osteoclast-like cell formation by sodium salicylate and indomethacin in mouse bone marrow culture. Jpn J Phamacol 1994:65:27-34
  26. Zhou D, Hughes B, King GJ, Histomorphornetnc and biochemical study of osteoclasts at orthodontic compression sites in the rat during indomethacin inhibition. Arch Oral BioI 1997;42:717-26 https://doi.org/10.1016/S0003-9969(97)00070-8
  27. Leroux P, Saffar JL. Dose-effect and evidence of escape of inhibition after indomethacin treatment in a synchronized model of bone resorption. Agents Actions 1993:38:290-4 https://doi.org/10.1007/BF01976223
  28. 신강섭, 이기수. 인도메타신이 개의 유치 치근흡수에 미치는 영향. 경희대학교 대학원 치의학과 박사논문, 2003