단순 만성 치주염 환자 및 2형 당뇨병을 가진 만성 치주염 환자의 치은조직에서 Matrix Metalloproteinase와 $TNF-{\alpha}$ 의 발현 양상 비교

Interrelationship of Matrix Metalloproteinase and $TNF-{\alpha}$ in Human Gingiva with Chronic Periodontitis associated to Type 2 Diabetes Mellitus

  • 김도훈 (경북대학교 치의학대학원) ;
  • 박의균 (경북대학교 치의학대학원) ;
  • 신홍인 (경북대학교 치의학대학원) ;
  • 조제열 (경북대학교 치의학대학원) ;
  • 서조영 (경북대학교 치의학대학원) ;
  • 이재목 (경북대학교 치의학대학원)
  • Kim, Doe-Heun (School of Dentistry, Kyungpook National University) ;
  • Park, Eei-Kyun (School of Dentistry, Kyungpook National University) ;
  • Shin, Hong-In (School of Dentistry, Kyungpook National University) ;
  • Cho, Je-Yeol (School of Dentistry, Kyungpook National University) ;
  • Suh, Jo-Young (School of Dentistry, Kyungpook National University) ;
  • Lee, Jae-Mok (School of Dentistry, Kyungpook National University)
  • 발행 : 2006.06.30

초록

치주질환의 병원균은 세포벽의 항원에 의하여 조직내 존재하는 mononuclear phagocytes가 활성화되어 cytokine들이 생성됨 으로써 치주 결체조직의 파괴를 진행시킨다. 이런 관련된 cytokine들은 순차적으로 상주하는 치은세포 및 대식세포가 Matrix metalloproteinase 합성을 하도록 유도하여 조직파괴를 시작한다. 이들 Matrix metalloproteinase중 MMP-2, MMP-9 (Gelatinase A,B)는 type IV collagen 및 변성된 interstitial collagen을 파괴하며 치주환자의 치은 열구액, 치은조직, 타액 네에서 높게 보고 되어왔다. 당뇨병은 치주질환의 위험요소중 하나로 달뇨 환자에서는 치주질환의 유병율이 일반인에 비해 높고 치주질환의 중증도도 더 심하여 진행도 빠르다고 알려져 있다. 그 병리 기전 중 하나로는 당뇨 환자에서는 치은 열구액 내 중성구 유래의 Matrix metalloproteinase의 활성 증가 및$TNF-{\alpha}$ 의 활성 증가가 추정되고 있다. 본 실험에서는 제2형 당뇨병 환자와 비당뇨 환자들에서 만성 치주염 부위의 치은 및 건강한 치은에서 염증매개체 중 하나인 MMP-2, MMP-9 및 $TNF-{\alpha}$ 의 발현에 대해 상호 비교 분석함으로서 염증, 혈당이 미치는 영향을 밝히고 제 2형 당뇨병 환자에서 심한 치주조직 파괴의 기전을 연구하고자 하였다. 경북대학교병원 치주과 내원환자 중 제2형 당뇨병 환자와 비당뇨 환자들 및 치주질환이 없는 건강인 대조군을 대상으로 여러 가지 환자요소, 임상 치주상태를 기록하고, 전신적으로 건강한 환자의 건강한 부위(n=8,Group 1), 전신적으로 건강한 환자으 만성 치주염 부위(n=8, Group 2), 제2형 당뇨병 환자의 만성 치주염 부위 (n=8,Group 3)에서 각각 변연치은을 채득하고 액화질소에 급속 동결하였다. Western blotting을 이용하여 각 조직 내 MMP-2, MMP-9 및 $TNF-{\alpha}$ 의 발현을 관찰, densitometer를 이용하여 상대적 발현을 정량, 각 조직의${\beta}-actin$을 이용하여 표준화하여 실험군과 대조군들의 평균치를 비교하였다. 비당뇨 환자들의 만성 치주염 부위 및 제2형 당뇨병 환자의 만성 치주염 부위에서 모두 건강 대조군에 비해 MMP-2와 MMP-9 의 발현이 증가되었다. 또한 MMP-2와 MMP-9는 2형 당뇨 환자의 만성 치주염 부위가 비당뇨 환자의 만성 치주염 부위보다 증가된 발현양상을 보였으며, $TNF-{\alpha}$ 발현 비교시 각 군간 유의성 있는 변화는 없었으나 2형 당뇨환자군에서 MMP-2 및 MMP-9의 증가와 함께 다소 증가 양상을 보였다. 결론적으로 본 실험에서 MMP-2 및 MMP-9의 증가가 만성 치주염 및 2형 당뇨 환자에서의 만성치주염에서 비당뇨환자 보다 MMP-2, MMP-9의 증가양상을 보여 주었으며 $TNF-{\alpha}$ 가 2형 당뇨환자의 만성치주염 진행과정에 기여인자로써 역할을 하는 것으로 생각된다.

키워드

참고문헌

  1. Yoshihio I, Fusanori N, Masatsugu N et al. The Effect of Antimicrobial Periodontal Treatment on Circulating Tumor Necrosis Factor-Alpha and Glycated Hemoglobin Level in Patients With Type 2 Diabetes. J Periodontol. 2001;72:774-778 https://doi.org/10.1902/jop.2001.72.6.774
  2. Ditto VJ, Overall CM and McCulloch C Proteolytic host cell enzymes in gingival-crevice fluid. Periodentology 2000 2003;31:77-104 https://doi.org/10.1034/j.1600-0757.2003.03106.x
  3. Ejeil AL, Igondjo-Tchen S, Ghomrasseni S et al. Expression of Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Metalloproteinases (TIMPs) in Healthy and Diseased Human Gingiva. Periodontol. 2003;74:188-195 https://doi.org/10.1902/jop.2003.74.2.188
  4. Ryan EM, Golub LM Modulation of Matrix metalloproteinase activities in periodontitis as a treatment strategy. Periodontology 2000. 2000:24:226-238 https://doi.org/10.1034/j.1600-0757.2000.2240111.x
  5. Souza AP, Gerlach RF and Line SRP Inhibition of human gingival gelatinases (MMP-2 and MMP-9) by metal salts. Dental Materials 2000;16:103-108 https://doi.org/10.1016/S0109-5641(99)00084-6
  6. Masanori N, Yoshinori Y, Kazusada Y and Yukikazu S Effects of $TNF-{\alpha}$ and prostaglandin E2 on the expression of MMPs in human periodontal ligament fibroblasts. Periodont Res. 2002;37: 167-176 https://doi.org/10.1034/j.1600-0765.2002.00656.x
  7. Meikle MC, Hembry RM, Holley J et al. Immunolocalization of matrix metalloproteinases and TIMP-1 (tissue inhibitor of metalloproteinases) in human gingival tissues from periodontitis patients. Periodont Res. 1994;29:118-126 https://doi.org/10.1111/j.1600-0765.1994.tb01100.x
  8. Chang Y-C, Yang S-F, Lai C-C, Liu J-Y and Hsieh Y-S Regulation of matrix metalloproteinase production by cytokines, pharmacological agents and periodontal pathogens in human periodontal ligament fibroblast cultures. Periodont Res. 2002;37:196-203 https://doi.org/10.1034/j.1600-0765.2002.00663.x
  9. Kiili M, Cox SW, Chen HW et al. Collagenase-2 (MMP-8) and collagenase-3(MMP-13) in adult periodontitis : Molecular forms and levels in gingival crevicular fluid and immunolocalisation in gingival tissue. J Clin Periodontol. 2002;29:224-232 https://doi.org/10.1034/j.1600-051x.2002.290308.x
  10. Desfaits A-C, Serri O and Renier G Normalization of Plasma Lipid Peroxides, Monocyte Adhesion and Tumor Necrosis $Factor-{\alpha}$ Production in NIDDM Patients After Gliclazide Treatment. Diabetes Care. 1998;21 487-493 https://doi.org/10.2337/diacare.21.4.487
  11. Erwin van der Zee, Everts V and Beertsen W Cytokines modulate routes of collagen breakdown. J Clin Periodontol. 1997;24:297-305 https://doi.org/10.1111/j.1600-051X.1997.tb00761.x
  12. Collin HL, Sorsa T, Meurman JH et al. Salivary Matrix Metalloproteinase (MMP-8) levels and Gelatinase(MMP-9) activities in patients with type 2 diabetes mellitus. J Periodont Res. 2000;35:259-265 https://doi.org/10.1034/j.1600-0765.2000.035005259.x
  13. Wall SJ, Sampson MJ, Levell N and Murphy G Cutaneous Biology. Elevated matrix metalloproteinase-2 and -3 production from human diabetic dermal fibroblasts. British Journal of Dermatology. 2003;149:13-16 https://doi.org/10.1046/j.1365-2133.2003.05262.x
  14. Grayson R, Douglas CWI, Heath J et al Activation of human matrix metalloproteinase 2 by gingival crevicular fluid and Porphyromonas gingivalis. J Clin Periodontol. 2003;30:542-550 https://doi.org/10.1034/j.1600-051X.2003.00301.x
  15. Birkendal-Hansen H Role of Matrix Metalloproteinases In Human Periodontal Diseases. J Periodontol. 1993;64:474-484 https://doi.org/10.1902/jop.1993.64.5.474
  16. Uemura S, Matsushita H, Li W et al. Diabetes Mellitus Enhances Vascular Matrix Metalloproteinase Activity. Role of Oxidative Stress. Circulation Research 2001;88:1291-1298 https://doi.org/10.1161/hh1201.092042
  17. Portik-Dobos V, Anstadt MP, Hutchinson J, Bannan M and Ergul A Evidence for a Matrix Metalloproteinase Induction/Activation System in Arterial Vasculature and Decreased Synthesis and Activity in Diabetes. Diabetes 2002;51:3063-3068 https://doi.org/10.2337/diabetes.51.10.3063
  18. Emrich LJ, Shlosman M and Genco RJ : Periodontal disease in non-insulin dependent diabetes mellitus. J Periodontol.1991;62:123-12 https://doi.org/10.1902/jop.1991.62.2.123
  19. Taylor GW, Burt BA, Becker MP et al. Non-insulin dependent diabetes mellitus and alveolar bone loss progression over 2 years. J Periodontol. 1998;69:76-83 https://doi.org/10.1902/jop.1998.69.1.76
  20. Collin H-L, Uusitupa M and Niskanen L Periodontal findings in elderly patient with non-insulin depedent diabetes mellitus. J Periodontol. 1998;69:962-966 https://doi.org/10.1902/jop.1998.69.9.962
  21. Salvi GE, Yalda B, Collins JG et al. Inflammatory Mediator Response as a Potential Risk Marker for Periodontal Diseases in Insulin-Dependent Diabetes mellitus patients. J Periodontol. 1997;68:127-135 https://doi.org/10.1902/jop.1997.68.2.127
  22. AAP Position Paper : Diabetes and peridontal disease. J Periodontol. 1999;70:935-949 https://doi.org/10.1902/jop.1999.70.8.935
  23. Yalda B, Offenbacher S and Collins JG Diabetes as a modifier of periodontal disease expression. Periodontology 2000. 1994;6:37-49 https://doi.org/10.1111/j.1600-0757.1994.tb00025.x
  24. Bissada NF, Manoucher-Pour M, Haddow M and Spagnulo PJ Neutrophil functional activity in juvenile and adult onset diabetic patients with mild and severe periodontitis. J Periodont Res. 1982;17:500-502 https://doi.org/10.1111/j.1600-0765.1982.tb02038.x
  25. Armitage GC Development of a classification system for periodontal disease and conditions. Ann Periodontol 19994:1-6
  26. Fusanori N, Yoshihiro I, Junji M et al. Periodontal Disease and Diabetes Mellitus: The Role of Tumor Necrosis $Factor-{\alpha}$ in a 2-way Relationship. Periodontol. 2003;74:97-102 https://doi.org/10.1902/jop.2003.74.1.97
  27. Quintero JC, Piesco NP, Langkamp HH, Bowen L and Agarwal S : LPS responsiveness in periodontal ligament cells is regulated by tumor necrosis factor-alpha. J Dent Res. 1995;74(11):1802-1811 https://doi.org/10.1177/00220345950740111401
  28. Persson L, Bergstr m J and Gustafsson A Effect of Tobacco Smoking on Neutrophil Activity Following Periodontal Surgery. J Periodontol. 2003;74:1475-1482 https://doi.org/10.1902/jop.2003.74.10.1475
  29. M hlman HR and Son SH Gingival Bleeding-a leading symptom in initial gingivitis. Helvitica Odontologica Acta. 1971;15:107-113
  30. Cho J-Y, Xing S, Liu X et al. Expression and activity of human Na+/I- symporter in human glioma cells by adenovirus-mediate gene delivery. Gene Therapy. 2000;7:740-749
  31. M kel M, Jalo T, Uitto V-J and Larjava H: Matrix Metalloproteinase (MMP-2 & MMP-9) of the oral cavity. J Dent Res. 73:1397-1406. 1994 https://doi.org/10.1177/00220345940730080201
  32. Page RC and Schroeder HE Parhogenesis of inflammatory periodontal disease. A summary of current work. Lab Invest.1976;34:235-249
  33. Ingman T, sorsa T, Lindy O, Koski H and Konttinen YT Multiple forms of gelatinases/type IV collagenases in saliva and gingival crevicular fluid of periodontitis patients. J Clin Periodontol. 1994;21:26-31 https://doi.org/10.1111/j.1600-051X.1994.tb00272.x
  34. Golub LM, Lee HM and Lehrer G Minocycline reduces gingival collagenolytic activity during diabetes. Preliminary observations and a proposed new mechanism of action. J Periodont Res. 1983;18:516-526 https://doi.org/10.1111/j.1600-0765.1983.tb00388.x
  35. Lalla E, Lamster IB, Feit M et al. Blockade of RAGE suppresses peiodontitis-associated bone loss in diabetic mice. J. Clin. Invest. 2000;105:1117-1124 https://doi.org/10.1172/JCI8942
  36. Ide M, Jagdev D, Coward PY et al. The short-term effects of treatment of chronic periodontitis on circulating levels of endotoxin, C-reactive protein, tumor necrosis $factor-{\alpha}$ and interleukin-6. J Peridontol. 2004;75:420-428 https://doi.org/10.1902/jop.2004.75.3.420
  37. Wahlgren J, Maisi P, Sorsa T et al. Expression and induction of collagenases(MMP-8 and -13) in plasma cells association with bone-destructive lesions. Journal of Pathology 2001;194:217-224 https://doi.org/10.1002/path.854
  38. Manoucher-Pour M, Spagnuolo PJ and Bissada NF Impaired neutrophil chemotaxis in diabetic patients with severe periodontitis. J Dent Res. 1981;60:729-730 https://doi.org/10.1177/00220345810600031101
  39. Uematsu S, Mogi M and Deguchi T $Interleukin(IL)-1{\beta}$ IL-6, Tumor Necrosis $Factor-{\alpha}$, Epidermal Growth Factor, and ${\beta}$ 2-microglobulin levels are elevated in gingival crevicular fluid during human orthodontic tooth movement. J Dent Res 1996;75:562-567 https://doi.org/10.1177/00220345960750010801
  40. Noda K, Ishida S, Inoue M et al. Production and activation of matrix metalloproteinase-2 in proliferative diabetic retinopathy. Invest Ophthalmol Vis Sci. 2003;44:2163-70 https://doi.org/10.1167/iovs.02-0662
  41. Nagase H and Woessner JF Jr. Matrix metalloproteinases. J Biol Chem. 1994;274:21491-21494 https://doi.org/10.1074/jbc.274.31.21491
  42. McLennan SV, Martell SK and Yue DK Effects of mesangium glycation on matrix metalloproteinase activities : possible role in diabetic nephropathy. Diabetes. 2002;51:2612-8 https://doi.org/10.2337/diabetes.51.8.2612
  43. Diamant M, Hanemaaijer R, Berheijen JH et al. Elevated matrix metalloproteinase-2 and -9 in urine, but not in serum, are markers of type 1 diabetic nephropathy. Diabet Med. 2001;18:423-4
  44. Jin M, Kashiwagi K, Iizuka Y, Tanaka Y, Imai M and Tsukahara S : Matrix metalloproteinases in human diabetic and non-diabetic vitreous. Retina. 21(1) : 28-33. 2001 https://doi.org/10.1097/00006982-200102000-00005
  45. Katsuki A, Sumida Y, Murashima S et al. Serum Levels of Tumor Necrosis $Factor-{\alpha}$ Are Increased in Obese Patients with Noninsulin-Dependent Diabetes Mellitus. J Clinical Endoclinology and Metabolism. 1998;83:859-862 https://doi.org/10.1210/jc.83.3.859
  46. Death AK, Fisher EJ, McGrath KCY and Yue DK High glucose alters matrix metalloroteinase expression in two key vascular cells : potential impact on artherosclerosis in diabetes. Arthersclerosis 2003;168:263-269 https://doi.org/10.1016/S0021-9150(03)00140-0
  47. Sakuta T, Tokuda M, Tamura M et al. Dual regulatory effects of interferon-alpha, -beta and -gamma interleukin-8 gene expression by human gingival fibroblasts in culture upon stimulation with lipopolysaccharide from Prevotell intermedia, interleukin-1alpha, or tumor necrosis factor-alpha. J Dent Res. 1998;77:1597-1605 https://doi.org/10.1177/00220345980770080701
  48. Ryan ME, Ramamurthy NS, Sorsa T and Golub LM MMP-mediated events in diabetes. Ann. NY Acad Sci. 1999;873:311-314
  49. Choi DH, Moon IS, Choi BK et al. Effects of sub-antimic robial dose doxycycline therapy on crevicular fluid MMP-8 and gingival tissue MMP-9, TIMP-1 and IL-6 levels in chronic periodontitis. J Periodont Res. 2004;39:20-26 https://doi.org/10.1111/j.1600-0765.2004.00696.x
  50. Dahan M, Nawrocki B, Elkaim R et al. Expression of matrix metallopoteinases in healhty and diseases human gingiva. J. Clin Periodontol 2001;28:128-136 https://doi.org/10.1034/j.1600-051x.2001.028002128.x