Expression of Matrix metalloproteinase-1 between Simple Chronic Periodontitis and Type 2 Diabetes associated Chronic Periodontitis on Protein level

단순만성치주염환자와 2형 당뇨환자의 만성치주염에서 Matrix metalloproteinase-1의 발현양상

  • Lee, Jae-Mok (Department of Periodontology, College of Dentistry, Kyungpook National University)
  • 이재목 (경북대학교 치과대학 치주과학교실)
  • Published : 2005.09.30

Abstract

The purpose of this study was to quantify and compare the level of MMP-1 in the healthy or inflamed gingival tissue of patients with or without type 2 diabetic mellitus. We investigated whether mean amount of MMP-1 was changed by chronic periodontitis and type 2 DM. Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was divided into the three group. Group 1(n=8) was clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2(n=8) was inflamed gingiva from patients with chronic periodontitis. Group 3(n=8) was inflamed gingiva from patients with chronic periodontitis and type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantitative analysis of MMP-1 was performed using a densitometer and statistically analyzed by ANOVA. MMP-1 was expressed in all samples and an increased MMP-1 level was observed in group 2 compared to group 1 and decreased MMP-1 level was found group 3 compared to group 2, but the differences among 3 groups were not statistically significant. In conclusion, this study demonstrated that MMP-1 levels of inflamed gingiva of systemically healthy patient(group 2) were higher than normal gingiva of systemically health patients and although the severity of gingival inflammation in group 2 and 3 were similar, MMP-1 expression was decreased in diabetic patients than systemically healthy periodontal patients.

본 연구의 목적은 전신적으로 건강한 치주질환자를 대조군으로 하여 제 2형 당뇨병을 동반한 치주질환자의 치은조직에서 MMP-1의 발현양상을 관찰, 비교하는 것으로 당뇨병을 동반한 경우 MMP-1의 발현양상이 변화되는지의 여부를 연구하였다. 경북대학교 병원 치주과에 내원한 환자 중 검사 및 수술에 동의한 환자로 전선 질환이 없고 부착 소설이 없거나 안정되어 있으며 치은 염증 소견이 없는 환자를 정상조직군, 임상적 치주낭 깊이가 5 mm 이상이고 방사선 사진상 치조골 소실이 분명한 환자를 만성 치주염 환자군, 심각한 전신적 합병증, 감염등의 위험요인이 없고 2형 당뇨병으로 진단받은 환자로서 만성 치주염으로 진단된 환지군 각 8명을 대상으로 하였다. 만성 치주염 환자와 당뇨병을 가진 만성 치주염 환자에서 치은 염증 소견을 보이는 치은조직을 채득하고 액화질소에 넣어 급속 동결고정시킨 후 MMP-1의 발현 양상을 western blot analysis를 통해 관찰하였고, densitometer를 이용하여 상대적 발현을 정량, 각 조직의 ${\beta}-actin$을 이용하여 표준화하여 각 군의 평균치를 비교하였다. 각 군 간의 차이를 one way ANOVA test로 분석하였다. 모든 군에서 분자량 53 kDa의 MMP-1에 상응하는 띠가 나타났으며 정량결과 전신적으로 건강한 치주염 환자군에서 MMP-1의 발현이 당뇨병을 동반한 치주염 환자군과 정상조직군의 치은조직에서보다 높게 나타났으나 통계적으로 유의한 차이는 나타나지 않았다. 치은염증의 존재시 MMP-1의 발현이 다소 증가됨을 관찰하였으나 통계적으로 유의한 수준은 아니였으며, 당뇨병을 동반한 치주염 환자군에서 전신적으보 건강한 치주염 환자군에서 보다 MMP-1의 발현이 감소되는 경향을 보였다.

Keywords

References

  1. Listgarten MA. : Pathogenesis of periodontitis. J Clin Periodontol 13:418-425. 1986 https://doi.org/10.1111/j.1600-051X.1986.tb01485.x
  2. Birkedal-Hansen H. : Role of matrix metalloproteinases in human periodontal diseases. J Periodontol 64:474-484. 1993
  3. Reynolds JJ, and Meikle MC. : Mechanisms of connective tissue matrix destruction in periodontitis. Periodontol 2000 14:144-157. 1997 https://doi.org/10.1111/j.1600-0757.1997.tb00195.x
  4. Westerlund D, Ingman T and Lukinmmaa P-L : Human neutrophil gelatinase and associated lipokalin in adult and localized juvenile periodontitis. J Dent Res 75: 1553-1563. 1996 https://doi.org/10.1177/00220345960750080601
  5. Nagase H and Woessner JF Jr. : Matrix metalloproteinases. J Biol Chem 274:21491-21494. 1999 https://doi.org/10.1074/jbc.274.31.21491
  6. Wincenti MP. : The matrix metal-lopro-teinase(MMP) and tissue inhibitor of metalloproteinase genes. Methods Mol Biol 151:121-148. 2001
  7. Nagase H : Activation mechanisms of matrix metalloproteinases. J Biol Chem 378:151-160. 1997
  8. Aiba T, Akeno N, Kawane T, Okamoto H and Horiuchi N. : Matrix metalloproteinases-1 and -8 and TIMP-1 mRNA levels in normal and diseased human gingivae. Eur J Oral Sci 104:562-569. 1996 https://doi.org/10.1111/j.1600-0722.1996.tb00142.x
  9. Kubota T, Nomura T, Takahashi T, and Kara K. : Expression of mRNA for matrix metalloproteinases and tissue inhibitors of metalloproteinases in periodontitisaffected human gingival tissue. Arch Oral Biol 41:253-262. 1994 https://doi.org/10.1016/0003-9969(95)00126-3
  10. Ingman T. Tervahartiala T and Ding Y : Matrix metalloproteinases and their inhibitors in gingival crevicular fluid and saliva of periodontitis patients. J Clin Periodontol 23: 1127-1132. 1996 https://doi.org/10.1111/j.1600-051X.1996.tb01814.x
  11. Soell M. Elkaim R. and Tenenbaum H. : Cathepsin C. matrix metalloproteinases. and their tissue inhibitors in gingiva and gingival crevicular fluid from periodontitis affected patients. J Dent Res 81: 174-178. 2002 https://doi.org/10.1177/154405910208100306
  12. Zee E. Everts V and Beertsen W. Cytokines modulate routes of collagen breakdown. Review with special emphasis on mechanism of collagen degradation in the periodontium and the burst hypothesis of periodontal disease progression. J Clin Periodontol 24:297-305. 1997 https://doi.org/10.1111/j.1600-051X.1997.tb00761.x
  13. Nelson RG, Shlossman M. and Budding LM. : Periodontal disease and NIDDM in Pima Indians. Diabetes Care 13:836-840. 1990 https://doi.org/10.2337/diacare.13.8.836
  14. Emrich LJ, Shlossman M and Genco RJ. : Periodontal disease in non-insulindependent diabetes mellitus. J Periodontol 62: 123-13. 1991 https://doi.org/10.1902/jop.1991.62.2.123
  15. Thorstensson H and Hugoson A. : Periodontal disease experience in adult long duration insulin-dependent diabetics. J Clin Periodontol 20:352-358. 1993 https://doi.org/10.1111/j.1600-051X.1993.tb00372.x
  16. Taylor GW, Burt BA. Becker MP. Genco RJ, Shlossman M. Knowler WC and Pettitt DJ. : Non-insulin dependent diabetes mellitus and alveolar bone loss progression over 2 years. J Periodontol 69: 76-83. 1998 https://doi.org/10.1902/jop.1998.69.1.76
  17. Collin HL, Uusitupa M and Niskanen L. : Periodontal findings in elderly patient with non-insulin dependent diabetes mellitus. J Periodontol. 69:962-966. 1998 https://doi.org/10.1902/jop.1998.69.9.962
  18. Wolf J. : Dental and periodontal conditions in diabetes mellitus. A clinical and radiographic study. Proc Finn Dent Soc 73(4-6 Suppl.): 1-56. 1977
  19. Seppala B, Seppala M and Ainamo J. : A longitudinal study on insulin-dependent diabetes mellitus and periodontal disease. J Clin Periodontol 20: 161-165. 1993 https://doi.org/10.1111/j.1600-051X.1993.tb00338.x
  20. Seppala B and Ainamo J. : A siteby-site follow-up study on the effect of controlled versus poorly controlled insulin-dependent diabetes mellitus. J Clin Periodontol 21: 161-165. 1994 https://doi.org/10.1111/j.1600-051X.1994.tb00297.x
  21. Taylor GW, Burt BA and Becker MP : Severe periodontitis and risk for poor glycemic control in patient with noninsulin-dependent diabetes mellitus. J Periodontol. 67: 1085-1093. 1996 https://doi.org/10.1902/jop.1996.67.10s.1085
  22. Taylor GW. : Bidirectional interrelationships between diabetes and periodontal diseases : An Epidomiologic Perspective. Ann Periodontol 6:99-112. 2001 https://doi.org/10.1902/annals.2001.6.1.99
  23. Cohen MP. : Non-enzymatic glvcosylation. Diabetes Annual 4:469-484. 1988
  24. Golub LM, Lee HM, and Lehrer G. : Minocycline reduces gingival collagenolytic activity during diabetes. J Perio Res 18:516-526. 1983 https://doi.org/10.1111/j.1600-0765.1983.tb00388.x
  25. AAP. Position Paper : Diabetes and periodontal disease. J Periodontol 70: 935-949. 1999 https://doi.org/10.1902/jop.1999.70.8.935
  26. Armitage GC. : Development of a classification system for periodontal diseases and conditions. Ann Periodontol 4: 1-6. 1999 https://doi.org/10.1902/annals.1999.4.1.1
  27. Armitage GC. : Periodontal findings in elderly patient with non-insulin dependent diabetes mellitus. J Periodontol. 69:962-966. 1998 https://doi.org/10.1902/jop.1998.69.9.962
  28. Muhlemann HR. and Son S. : Gingival sulcus bleeding a leading symptom in initial gingivitis. Helv Odontol Acta 15: 107. 1971
  29. Cho J-Y. Xing S. Liu X. Buckwalter TLF, Hwa L. Sferra TJ. Chiu IM and Jhiang SM. : Expression and activity of human $Na^+/I^-$ symporter in human glioma cells by adenovirus-mediated gene delivery. Gene Therapy 7:740-749. 2000
  30. Tervonen T. and Oliver RC. : Long-term control of diabetes mellitus and periodontitis. J Clin Periodontol 20:431-435. 1993 https://doi.org/10.1111/j.1600-051X.1993.tb00384.x
  31. Grossi SG. Zambon JJ. Ho AW, Koch G. Dunford RG. Machtei EE. Norderyd OM and Genco RJ. : Assessment for risk of periodontal disease. I. Risk indicators for attachment loss. J Periodontol 65: 260-267. 1994 https://doi.org/10.1902/jop.1994.65.3.260
  32. Ueta E. Osaki T. Yoneda K and Yamamoto T. : Prevalence of diabetes mellitus in odontogenic infections and oral candidacies: an analysis of neutrophil suppression. J Oral Patho Med 22: 168-194. 1993 https://doi.org/10.1111/j.1600-0714.1993.tb01051.x
  33. Chappey O. Dosquet C. Wautier MP and Wautier JL. : Advanced glycation end products. oxidant stress and vascular lesions. Eur J Clin Invest 27:97-108. 1997 https://doi.org/10.1046/j.1365-2362.1997.710624.x
  34. Feener EP and King GL. : Vascular dysfunction in diabetes mellitus. Lnacet 350: S19-S113. 1997
  35. Schmidt AM. Weidman E. Lalla E. Yan SD. Hori O. Cao R, Brett JG and Lamster IB. : Advanced glycation end products induce oxidant stress in gingiva: a potential mechanism underlying accelerated periodontal disease associated with diabetes. J Periodont Res 31: 508-515. 1996 https://doi.org/10.1111/j.1600-0765.1996.tb01417.x
  36. Sorsa T. Ding Y and Ingman T. : Cellular source, activation and inhibition of dental plaque collagenase. J Clin Periodontol 22:709-717. 1995 https://doi.org/10.1111/j.1600-051X.1995.tb00831.x
  37. Nomura T, Takahashi T and Hara K. : Expression of TIMP-1, TIMP-2 and collagenase mRNA in periodontitis-affected human gingival tissue. J Perio Res 28:354-362. 1993 https://doi.org/10.1111/j.1600-0765.1993.tb01079.x
  38. Meikle MC, Hembry RM. Holley J, Horton C, McFarlane CG and Reynolds JJ. : Immunolocalization of matrix metalloproteinases and TIMP-1 in human gingival tissues from periodontal patients. J Periodont Res 29: 118-126. 1994 https://doi.org/10.1111/j.1600-0765.1994.tb01100.x
  39. Death AK. Fisher EJ, McGrath KCY and Yue DK. : High glucose alters matrix metalloproteinase expression in two key vascular cells: potential impact on atherosclerosis in diabetes. Athero-sclerosis 168: 263-269. 2003 https://doi.org/10.1016/S0021-9150(03)00140-0
  40. 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 51: 3063-3068. 2002 https://doi.org/10.2337/diabetes.51.10.3063
  41. Rittie L, Berton A. Monboisse JC, Hornebeck W and Gillery P. : Decreased contraction of glycated collagen lattices coincides with impaired matrix metalloproteinase production. Biochemical and Biophysical Research Communications 264:488-492. 1999 https://doi.org/10.1006/bbrc.1999.1519