• Restorative Dentistry and Endodontics
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• v.26 no.3
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• pp.191-199
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• 2001

목적 - 기질금속단백분해효소(matrix metalloproteinase)는 조직의 염증 및 치유과정에서 숙주세포에서 생성, 분비되어 세포외기질(extracellular matrix)의 분해에 작용한다. 다양한 염증반응에 기질금속단백분해효소가 중요한 역할을 하는것으로 보고되고 있으나 치수 및 치근단 질환에서의 그 역할은 거의 알려져 있지 않은 상태이다. 본 연구에서는 염증이 있는 사람의 치수 및 치근단 조직을 채취하여 Enzymeimmunoassay 및 면역조직화학적 검색을 통해 제1형, 2형, 3형 기질금속단백분해효소의 수준 및 그 분포를 측정하여 치수 및 치근단 병소에서 이 효소의 작용을 알아보는 것을 목적으로 한다. 방법 - 연구재료는 근관치료를 위해 서울대학교 병원 치과 진료부 보존과에 내원한 환자를 대상으로 34개의 치아에서 통상의 근관치료 중 발수한 치수조직과 치근단 수술중 얻은 치근단 병소(n=10)를 이용하였다. 치수는 발수 전에 임상진단을 통해 급성 치수염(n=12), 만성 치수염(n=12), 정상 치수(n=10)로 구분하고 정상치수로 진단된 것을 대조군으로 설정하였다. 채취된 표본은 둘로 나누어 절반은 30분 이내에 5$\mu\textrm{m}$ 두께로 동결절단을 시행하여 조직표본을 제작하였고 deep freezer에 보관하였다가 헤마톡실린-에오신 염색 및 면역조직화학적 검색을 시행하였다. 나머지 조직은 ELISA를 위해 액체 질소에 보관하였다. ELISA를 시행하기전 표본의 단백질 정량을 시행하여 모든 표본의 단백질 양을 50mg/$\mu\textrm{l}$로 일치시키고 Amersham사의 ELISA kit를 사용하여 제1형, 2형, 3형의 기질금속단백분해효소의 양을 측정하였으며 그 결과를 Mann-Whitney U test를 사용하여 각 군간의 통계학적 유의성을 검증하였다. 결과 1. ELISA의 결과 제1형 기질금속단백분해효소의 농도는 모든 실험군에서 대조군보다 유의성있게 높게 나타났다.(p<.05). 또한 급성치수염군의 제1형 기질금속단백분해효소의 농도가 다른 실험군보다 유의성있게 높았다(p<.05). 2. 제2형 기질금속단백분해효소의 경우 급성치수염군과 대조군에서만 유의성있는 차이를 보였다(p<.05). 3. 제3형 기질금속단백분해효소의 경우 급성치수염군에서 대조군이나 만성치수염군보다 유의성 있는 높은 수치를 보였다(p<.05). 4. 면역조직화학검색 결과 염증성 치수에 존재하는 급성 및 만성염증세포 주위로 기질금속단백분해효소에 대한 면역 반응이 존재하였으며 주로 제1형과 제3형 기질금속단백분해효소의 경우 대식세포 및 림파구 주위로 강한 발색제의 침윤양상이 관찰되었다. 5. 치근단병소의 면역조직화학적 검색 결과 만성염증 세포 주변으로 미약한 발색제의 침윤양상이 관찰되었다.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.579-590
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• 2006

1. 연구배경 교원질 분해작용을 하는 호중구의 세포질 효소인 기질금속단백분해효소-8은 치주질환, 류마티스 관절염, 그리고 궤양결장염과 같은 염증성 질환에서 농도가 증가한다고 알려져 있다. 최근에는 A. actinomycetemcomitans의 leukotoxin이 사람호중구에서 기질금속단백분해효소-8의 분비를 유도하는 것이 보고되었다. 이 연구의 목적은 선천면역 체계에서 세포표면 항원무리14, Toll-like 수용기, 그리고 $NF-{\kappa}$ B경로를 통하여 A. actinomycetemcomitans의 지질다당질로 유도된 기질금속단백분해효소-8의 분비 여부와 세포기전을 알아보고자 하였다. 2. 연구재료 및 방법 건강한 개인 제공자(남자 13명, 여자 3명)로부터 얻은 개개인의 20ml 말초혈액을 제조사의 지침에 따라 호중구를 추출한 후 항세포표면 항원무리14와 함께 $4^{\circ}C$에서 30분간 전배양 한 후, $37^{\circ}C$에서 9시간 동안 배양시켰다. 추출한 호중구에 Toll-like 수용기 억제제 또는 $NF-{\kappa}$ B억제제인 TPCK를 첨가한 후 $37^{\circ}C$에서 1시간 동안 전배양하고 $37^{\circ}C$에서 9시간 동안 배양시켰다. 호중구에 세포뼈대 억제제인 cholchicine, nocodazole, demecolcine, 그리고 cytochalasin B를 A. actinomycetemcomitans의 지질다당질과 함께 $37^{\circ}C$에서 9시간 동안 배양시켰다. 기질금속단백분해효소-8 분비량은 효소면역측정법을 통해 결정하였다. 통계처리는 일원배치 분산분석법을 이용하였다(p<0.05). 3. 결과 A. actinomycetemcomitans 지질다당질은 기질금속단백분해효소-8의 분비를 증가시켰다. 기질금속단백분해효소-8의 분비는 항세포표면 항원무리14에 의해서 억제되었지만, 항 Toll-like 수용기2, 항 Toll-like 수용기4 항체는 억제시키지 못했다. $NF-{\kappa}$ B 억제제는 A. actinomycetemcomitans의 지질다당질로 유도된 $NF-{\kappa}$ B 결합 활성도와 기질금속단백분해효소-8 분비를 억제하였다. 미세섬유 중합반응 억제제는 A. actinomycetemcomitans의 지질다당질로 유도된 기질금속단백분해효소-8의 분비를 억제시켰으나, 미세관 중합반응억제제는 억제시키지 못했다. 4. 결론 위의 연구결과를 종합하여 볼 때, 기질금속단백분해효소-8은 A. actinomycetemcomitans의 지질다당질로 유도되며, 세포표면 항원무리-$NF-{\kappa}$ B 경로를 통하여 분비되고, 이 분비 과정은 미세섬유 계통이 관여하는 것으로 보인다.

• Tuberculosis and Respiratory Diseases
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• v.59 no.5
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• pp.517-521
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• 2005

Background : The balances of the proteinases and antiproteinases system have been implicated in the pathogenesis of various exudative pleural effusions. The aim of this study was to examine the matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels in exudative pleural effusions. Methods : The study included 33 tuberculous effusions, 17 malignant, and 5 transudates. The pleural levels of MMP-1 and TIMP-1 were determined using a commercially available ELISA assay. Results : The group of tuberculous effusions showed higher pleural MMP-1 levels than the malignant and transudates. The pleural TIMP-1 levels of the tuberculous and malignant effusions were higher than the transudates. Conclusion : Elevated pleural MMP-1 and TIMP-1 levels were found in tuberculous effusions.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.231-235
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• 2005

Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-dependent enzymes responsible for proteolytic degradation of extracellular matrix components such as base membrane or interstitial stroma. MMPs play an important role in a variety of physiological and pathological tissue remodeling processes, including wound healing, embryo implantation, tumor invasion and metastasis. Since MMP-9 (gelatinase B) has unique ability to cleave type IV collagen, gene expression of MMP-9 has been focused on as a pharmacological target. Flavonoids are a class of compounds that are widely spread in plants. In the coures of screening for the suppressors of MMP-9 gene expression from natural products, Metasequoia glyptostroboides was selected. Six flavonoids, sciadopitysin, isoginkgetin, bilobetin, 2,3-dihydrohinokiflavone, luteolin and apigenin were purified as suppressors of MMP-9 gene expression from M. glyptostroboides. The suppressing activity of the isolated flavinoids on the MMP-9 gene expression was measured by gelatin zymography and Nothern blot analysis.

• Radiation Oncology Journal
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• v.21 no.1
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• pp.66-74
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• 2003

Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.974-979
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• 2009

Proliferation of vascular smooth muscle cell(VSMC) is one of the key features in onset of atherosclerosis and restenosis after vascular surgery such as stent implant. Atherosclerotic plaques are usually composed of collagen, elatsin and smooth muscle cells. Release of matrix metalloproteinases(MMPs) is considered to have correlation with development of atherosclerotic plaques. Based on the hypothesis that MMP inhibition would be helpful in the treatment of atherosclerosis, we investigated inhibition of MMP activity and migration of TNF-$\alpha$-induced human aortic smooth muscle cell(HASMC) by Cinnamomi Ramulus(CC). The result from gelatin zymography showed that CC inhibited MMP-9 activity in a dose-dependent manner. In addition, CC considerably inhibited the migration of HASMC induced by TNF-$\alpha$, while it showed little cytotoxic effect on HASMC. These results suggest that CC can be a potential anti-atherosclerotic agent through inhibition of MMP-9 activity and SMC migration.

• The korean journal of orthodontics
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• v.36 no.2 s.115
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• pp.91-102
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• 2006

Tooth eruption requires remodeling of surrounding tissues. This study was aimed to investigate the effect of indomethacin on the dental follicle and paradental tissues during tooth eruption by observing the distribution and expression of MMP by the immunohistochemical method. Ten mongrel dogs of ten to twelve weeks old were divided into 5 groups; four experimental groups administered indomethacin 2 mg/Kg/day and 8 mg/Kg/day orally 2 times a day for 14 days and 7 days respectively, and the control group was administered a placebo. Permanent teeth before eruption and their surrounding tissues were selected and excised. H&E staining and immunohistochemical stainings of MMP-3 and -9 were performed and examined under the light microscope. Osteoclasts, osteoblasts, periodontal ligament cells, ameloblasts and odontoblasts of the control group all expressed MMP-3 and -9. In the experimental group, osteoclasts, osteoblasts and periodontal ligament cells showed reduced expression of MMP-3 and -9. Magnitude of MMP reduction In the experimental group showed a time and dose of indomethacin administration dependent manner. These results show that indomethacin inhibited MMP-3 and -9 expression in the dental follicle and surrounding tissues and suggest that when indomethacin is administered for long periods, tooth eruption could be delayed.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.580-586
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• 1989

Penicillium sp. CB-20 was selected for strong polygalacturonase activity among various strains of molds found in soil. The optimum pH for the enzyme activity was 5.0 and optimum temperature was 4$0^{\circ}C$. The enzyme was relatively stable in acidic condition and unstable by heat treatment. The activation energy, Km and V$_{max}$ for the polygalacturonase were 2.499 Kcal/mol, 2.13$\times$10$^{-2}$mol/l, and 104.17 $\mu$mol/min. The activity of polygalacturonase was inhibited by Ag$^{+}$, Cu$^{++}$, Pb$^{++}$, Fe$^{+++}$, $Ca^{++}$, Na$^+$, Mn$^{++}$. The enzyme can be inactivated by the treatment ethylenediamintetra acetic acid, 2,4-dinitrophenol and $H_2O$$_2$. The results indicate the possible involvement of histidine, chelate and terminal amino group as active site. The enzyme was endo-type polygalacturonase.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.372-384
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• 2005

The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.71-79
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• 1988

For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.