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

국문 초록 (Abstract)

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.

다국어 초록 (Multilingual 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.

참고문헌 (Reference)

1 "and tissue inhibitor of metalloproteinase genes" Wincenti MP 121-148, 2001

2 "and their tissue inhibitors in gingiva and gingival crevicular fluid from periodon- titis affected patients" Cathepsin C 174-178, 2002

3 "activation and inhibition of dental plaque collagenase" 709-717, 1995

4 "Vascular dys- function in diabetes mellitus" -350, 1997

5 "Severe periodontitis and risk for poor glycemic control in patient with non- insulin-dependent diabetes mellitus" 1085-1093, 1996

6 "Role of matrix me- talloproteinases in human periodontal diseases" 64 : 474-484, 1993

7 "Prevalence of diabetes mel- litus in odontogenic infections and oral candidacies an analysis of neutrophil suppression" 168-194, 1993

8 "Position Paper : Diabetes and periodontal disease" 70 : 935-949, 1999

9 "Periodontal findings in elderly patient with non-insulin dependent diabetes mellitus" 962-966, 1998

10 "Periodontal disease in non-insulinde- pendent diabetes mellitus" 123-13, 1991

11 "Periodontal disease and NIDDM in Pima Indians" 836-840, 1990

12 "Perio- dontal disease experience in adult long duration insulin-dependent diabetics" 352-358, 1993

13 "Pathogenesis of perio- dontitis" 418-425, 1986

14 "Non-insulin dependent dia- betes mellitus and alveolar bone loss progression over 2 years" 76-83, 1998

15 "Non-enzymatic glycosyla- tion" 4 (4): 469-484, 1988

16 "Minocycline reduces gingival collageno- lytic activity during diabetes" 516-526, 1983

17 "Mecha- nisms of connective tissue matrix de- struction in periodontitis" 14 : 144-157, 2000

18 "Matrix metalloproteinases and their inhibitors in gingival crevicular fluid and saliva of periodontitis patients" 1127-1132, 1996

19 "Matrix metallopro- teinases-1 and -8 and TIMP-1 mRNA levels in normal and diseased human gingivae" ake (ake): 562-569, 1996

20 "Ma- trix metalloproteinases" 21491-21494, 1999

21 "Long- term control of diabetes mellitus and periodontitis" 20 : 431-435, 1993

22 "Immunolocalization of matrix metalloproteinases and TIMP-1 in hu- man gingival tissues from periodontal patients" 118-126, 1994

23 "Human neutrophil gelati- nase and associated lipokalin in adult and localized juvenile periodontitis" 1553-1563, 1996

24 "High glucose alters matrix metalloproteinase expression in two key vascular cells potential impact on atherosclerosis in diabetes" 263-269, 2003

25 "Gingival sulcus bleeding a leading symptom in initial gingivitis" 1971

26 "Expression of mRNA for matrix metalloproteinases and tissue inhibitors of metalloproteinases in periodontitisaf- fected human gingival tissue" 253-262, 1994

27 "Expression of TIMP-1 TIMP-2 and colla- genase mRNA in periodontitis-affected human gingival tissue" 354-362, 1993

28 "Expression and activity of human Na+/I- symporter in human glioma cells by adenovirus-mediated gene delivery" 740-749, 2000

29 "Evi- dence for a Matrix Metalloproteinase induction/activation system in arterial vasculature and decreased synthesis and activity in diabetes" 3063-51 3068, 2002

30 "Development of a classi- fication system for periodontal diseases and conditions" 4 : 1-6, 1999

31 "Dental and periodontal condi- tions in diabetes mellitus" 4 1-6 56, 1977

32 "Decreased contraction of glycated collagen lattices coincides with impaired matrix metal- loproteinase production" 488-492, 1999

33 "Decreased contraction of glycated collagen lattices coincides with impaired matrix metal- loproteinase production" 488-492, 1999

34 "Cyto- kines modulate routes of collagen break- down Review with special emphasis on mechanism of collagen degradation in the periodontium and the burst hypothesis of periodontal disease progression" 297-305, 1997

35 "Bidirectional interrelation- ships between diabetes and periodontal diseases" 99-112, 2001

36 "Assessment for risk of periodontal disease Risk indicators for attachment loss J Periodontol 65" 260-267, 1994

37 "Advanced glycation end products induce oxidant stress in gin- giva a potential mechanism underlying accelerated periodontal disease associat- ed with diabetes" 508-515, 1996

38 "Advanced glycation end products" 27 : 97-108, 1997

39 "Activation mechanisms of matrix metalloproteinases" 151-160, 1997

40 "A site- by-site follow-up study on the effect of controlled versus poorly controlled insulin-dependent diabetes mellitus" 161-165, 1994

41 "A longitudinal study on insulin-depen- dent diabetes mellitus and periodontal disease" 161-165, 1993