• Title/Summary/Keyword: matrix metalloproteinases

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Cloning of a matrix metalloproteinase cDNA from Scylliorhinus torazame (두툽상어 matrix metalloproteinase 유전자 cDNA의 클로닝)

  • Kim, Jon Won;Cho, Won Jin;Chun, Kwang Ho;Kim, Kyu-Won;Kim, Yung-Jin;Lee, Sang-Jun;Shin, Hae-Ja;Lim, Woon Ki
    • Journal of Life Science
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    • v.8 no.3
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    • pp.235-240
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    • 1998
  • Matrix metalloproteinases(MMPs) are a group of zinc enzymes responsible for degradation of the matrix components such as collagen and proteoglycans in normal embryogenesis and remodeling and in many disease processes such as arthritis, cancer, periodontitis, and osteprocess. Genetically distince MMPs have been characterized and their genes have been cloned thus far from a variaty of species but not from fishes. In this stydy, a mmp cDNA was cloned by using RT-PCR(reverse transcriptase dependent polymerase chain reaction) from Scylliorhinus toraxzame(shark), agroup of cartilaginous fish, abundant in the coast of Pusan, Korea. It has 74% base homologue with membrane type matrix matalloproteinase-3 genes(mt3-mmps) from human, rat and chick, and also shows more than 90% residue homologue with them. In addition, it has cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif, which are present in MMPs. This result indicates that cDNA fragment cloned here may be mt3-mmp or its analogous gejne cDNA fragment of Scylliorhinus torzame.

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Peptide Hydrolysates from Astragalus membranaceus Bunge Inhibit the Expression of Matrix Metalloproteinases in Human Dermal Fibroblasts

  • Park, Sun Ki;Van Hien, Pham;Van Luong, Hoang;Yan, Shao-Wei;Byun, Sang Yo
    • KSBB Journal
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    • v.29 no.5
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    • pp.380-384
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    • 2014
  • Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

Protective Effects of Prunus persica Flesh Extract (PPFE) on UV-Induced Oxidative Stress and Matrix Metalloproteinases Expression in Human Skin Cells

  • Park, Hyen-Joo;Park, Kwang-Kyun;Hwang, Jae-Kwan;Chung, Won-Yoon;Kim, Gi-Dae;Lee, Min-Ai;Lee, Sang-Kook
    • Natural Product Sciences
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    • v.18 no.1
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    • pp.52-59
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    • 2012
  • In our continuous efforts to procure the active materials from natural products in the protective effects of oxidative stress or UV damage to skin cells we found the Prunus persica flesh extract (PPFE) is considerable to meet the demand to protect the skin damage. PPFE attenuated cell damage induced by hypoxanthine-xanthine oxidase in cultured human keratinocytes, indicating that PPFE has the potential of the scavenging effect of reactive oxygen species (ROS) in human skin cell. Moreover, PPFE significantly suppressed UVA-induced ROS production determined by the oxidation of 2,7-dichlorodihydrofluorescein diacetate (DCFH) using FACS analysis. Additional study revealed that UVA irradiation of HaCaT human keratinocytes increased the gelatinolytic activities of matrix metalloproteinase-2, and -9 (MMP-2, -9) and mRNA expression of MMP-9 analyzing by a real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and these events were significantly suppressed by the treatment with PPFE. These results suggest that PPFE might be applicable as natural ingredients for skin antiaging agents via UV-induced ROS scavenging activity and suppression of MMP expression in the skin cells.

Inhibition of the expression on MMP-2, 9 and morphological changes via human fibrosarcoma cell line by 6,6'-bieckol from marine alga Ecklonia cava

  • Zhang, Chen;Li, Yong;Shi, Xiujuan;Kim, Se-Kwon
    • BMB Reports
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    • v.43 no.1
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    • pp.62-68
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    • 2010
  • Matrix Metalloproteinases (MMPs) are a family of zinc-endopeptidases which can degrade extracellular matrix (ECM) components and play important roles in a variety of biological and pathological processes. 6,6'-bieckol isolated and characterized from an edible marine brown alga Ecklonia cava (EC), according to the comprehensive spectral analysis of MS and NMR data. Here the influence of 6,6'-bieckol on expressions of MMPs was examined by zymography and western blot analysis via human fibrosarcoma cell line (HT1080). It is shown that 6,6'-bieckol significantly down regulated the expressions of MMP-2 and -9 in dose-dependent manner. The influence of 6,6'-bieckol on the cell viability and cell behavior of HT1080 cells were also investigated, our dates shown that it suppressed the migration and 3D culture in HT1080 cells. Meanwhile, we explored several signal pathways which may contribute to this process, and found the suppressing of MMPs expressions in HT1080 cells might be due to the suppression of NF-${\kappa}B$ signal pathway.

Identification of the active components inhibiting the expression of matrix metallopeptidase-9 by TNFα in ethyl acetate extract of Euphorbia humifusa Willd

  • Ahn, Seunghyun;Jung, Hyeryoung;Jung, Yearam;Lee, Junho;Shin, Soon Young;Lim, Yoongho;Lee, Young Han
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.367-374
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    • 2019
  • Euphorbia humifusa Willd (EuH), called Ttang-Bin-Dae in Korea, is a traditional medicinal plant widely used for its anti-inflammatory and antiviral activity. Ethyl acetate (EA) extracts of EuH (EA/EuH) inhibit invasion and metastasis by inhibiting tumor necrosis factor TNFá-induced matrix metalloproteinases (MMP)-9 expression in human breast cancer cells. However, the bioactive components of EA/EuH mediating the inhibition of MMP-9 expression have not been identified. In the present study, three bioactive constituents of EA/EuH were isolated using high-performance liquid chromatography. Nuclear magnetic resonance spectroscopy revealed isoquercetin, avicularin, and astragalin as the bioactive compounds responsible for preventing TNFα-induced MMP-9 mRNA expression in breast cancer cells. These findings suggest that isoquercetin, avicularin, and astragalin could be used as valuable anti-metastatic agents against metastatic cancers.

Secretory Differentiation of Hamster Tracheal Epithelial Cells Increases Activation of Matrix Metalloproteinase-2

  • Shin, Chan-Young;Lee, Woo-Jong;Park, Kyu-Hwan;Ryu, Jae-Ryun;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.12 no.1
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    • pp.1-8
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    • 2004
  • In chronic airway inflammatory diseases such as asthma and chronic bronchitis, it has been suggested that matrix metalloproteinases secreted from infiltrating neutrophil contribute the pathogenesis of the disease and have been a focus of intense investigation. We report here that hamster tracheal surface epithelial goblet cells (HTSE cells) produce matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2). Matrix metalloproteinase activities were investigated using [$^3H$]collagen-digestion assay and gelatin zymography. The subtype of matrix metalloproteinases expressed from HTSE cells was MMP-2 (gelatinase A), which was determined by Western blot with various subtype selective anti-matrix metalloproteinase antibodies. The MMP-2 and TIMP-2 cDNAs from HTSE cells were partially cloned by RT-PCR and they reveal more than 90% of sequence homology with those from human, rat and mouse. The collagenolytic activity was increased with the secretory differentiation of the HTSE cell and it was found that zymogen activation was responsible for the increased MMP-2 activity in HTSE cells. The results from the present study suggest that the metaplastic secretory differentiation of airway goblet cells may affect chronic airway inflammatory process by augmenting the zymogen activation of MMP-2.

Roles of Matrix Metalloproteinases on Intracellular Staphylococcus aureus Growth in Bronchial Epithelial Cell (황색포도알균의 감염에 따른 세포 내에서의 균의 증식과 Matrix Metalloproteinase (MMP)의 역할)

  • Min, Bo Ram;Lee, Young Mi;Park, Jae Seok;Choi, Won-Il;Kwon, Kun Young
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.1
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    • pp.22-27
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    • 2008
  • Background: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity. Methods: In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor. Results: The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased. Conclusion: MMPs may mediate the invasion of S. aureus into epithelial cells.

Fructose 1.6-diphosphate Prevents Cyclooxygenase-2 and Matrix Metalloproteinases Expression by Inhibition of UVB-induced Signaling Cascades in HaCaT Keratinocytes (인체각질형성세포에서 Fructose 1,6-diphosphate의 자외선에 의해 유도되는 Cyclooxygenase-2 and Matrix Metalloproteinases의 발현억제기전)

  • Soo Mi, Ahn;Ji Hyun, Kim;Byeong Gon, Lee;Soo Hwan, Lee;Ih Seoup, Chang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.247-251
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    • 2004
  • UV radiation exerts various influences in the skin, including photoaging and inflammation (1). The MMPs (Matrix metalloproteinases), which are induced by UV irradiation, can degrade matrix proteins, and these results in a collagen deficiency in photodamaged skin that leads to skin wrinkling. It has been known that the production of PGE$_2$ stimulates MMPs expression, and inhibits procollagen (2). Thus, it is possible that the induction of MMPs and the inhibition of matrix protein synthesis by UV -induced PGE$_2$ may play some role in UV-induced collagen deficiency in photoaged skin. Fructose-1,6-diphosphate (FDP), a glycolytic metabolite, is reported to have cytoprotective effects against ischemia and postischemic reperfusion injury of brain and heart, presumably by augmenting anaerobic carbohydrate metabolism (3). And also, FDP significantly prevent skin aging by decreasing facial winkle compared with vehicle alone after 6 months of use. We studied the mechanism of anti-aging effect of FDP on UVB-irradiated HaCaT keratinocyte model. FDP has protective role in UVB injured keratinocyte by attenuating prostaglandin E$_2$ (PGE$_2$) production and COX-2 expression. And FDP also suppressed UVB-induced MMP-2 expression. Further, to delineate the inhibition of UVB-induced COX-2 and MMPs expression with cell signaling pathways, treatment of FDP to HaCaT keratinocytes resulted in marked inhibition of UVB-induced phosphorylation of ERK1/2, JNK. It also prevents UV induced NFB translocation, which are activated by cellular inflammatory signal. Our results indicate that FDP has protecting effects in UV-injured skin aging by decreasing UVB-induced COX-2 and MMPs expression, which are possibly through blocking UVB-induced signal cascades.

Antioxidant and Metalloproteinase Inhibitory Activities of Ethanol Extracts from Lespedeza cuneata G. Don (야관문 에탄올 추출물의 항산화 및 Metalloproteinase 저해 활성)

  • Shin, Yong Ha;Song, Chang-Khil
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.263-268
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    • 2017
  • BACKGROUND: Lespedeza cuneata G. Don is a well-known medicinal plant. In this study, the biological activities of L. cuneata extracts were investigated. METHODS AND RESULTS: L. cuneata shoot was extracted with 30% ethanol and further fractionated with organic solvents. Total phenolic and flavonoid content, antioxidant activity, and matrix metalloproteinase inhibition effect of the extract and fractions were measured. Among the tested extract and fractions, the highest contents of total phenolic and flavonoids were found in ethyl acetate fraction (117.8 mg GAE/g and 35.9 mg QE/g, respectively). Ethyl acetate fraction showed the highest DPPH and ABTS radical scavenging activity, and the antioxidant activity of the other fractions followed the order n-hexane fraction>ethanol extract>methyl chloroform>n-butanol fraction. Inhibitory effect on the expression of matrix metalloproteinases (MMP1 and MMP3) was highest in the fraction of ethyl acetate, and n-butanol fraction also significantly inhibited the expression of MMP3. Antioxidant activities of L. cuneata extracts were significantly positively related to their phenolic and flavonoid content. CONCLUSION: Ethyl acetate fraction of L. cuneata ethanol extract showed potent antioxidant and matrix metalloproteinases inhibitory activities. Those activities might be related to the high total phenolic and flavonoid content of the extract.

Analysis of Matrix Metalloproteinase-9 Expression in Renal Cell Carcinoma (신세포암종에서 Matrix Metalloproteinase-9 발현의 분석)

  • Kim, Ji-Yoon;Park, Tong-Choon
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.82-89
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    • 2006
  • Background: Matrix metalloproteinases (MMPs) are involved in the degradation of the extracellular matrix, which is an important step in tumor invasion and metastasis. A positive correlation between the expression of MMP-9 and aggressive behavior of renal cell carcinomas (RCCs) has been reported. MMP-9 expression in RCCs and adjacent normal kidney tissues were examined in this study. Materials and Methods: Twenty-five patients pathologically diagnosed as clear cell RCCs, from specimens obtained at radical nephrectomy, between May 2003 and December 2004 were enrolled in this study. MMP-9 activity was estimated using gelatin zymography, and quantified using a laser densitometer. The results were compared with clinicopathological characteristics. Results: The expression of MMP-9 was significantly elevated in the RCC compared with non-tumor kidney specimens (p<0.01). The levels of MMP-9 expression in the RCC patients with large tumors (>4 cm) or vascular invasion were significantly higher than in those without these clinical manifestations (p<0.01). There were also significant differences in the expression of MMP-9 among T stages (p<0.01). The tissue MMP-9 level was the highest in nuclear grade 4, but there was no statistical significance between the histological grades (p=0.17). Conclusions: These results suggest that enhanced MMP-9 expression contributes to carcinogenesis and tumor progression in the later stages of RCC.

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