• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.29-36
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• 2012

Osteosarcoma is the most common primary malignancy of bone, and patients often develop pulmonary metastasis. The mechanisms underlying osteosarcoma metastasis remain to be elucidated. Recently, anti-inflammatory agents were shown to be useful in the treatment of tumor progression. We previously isolated a natural anti-inflammatory peptide from the seahorse Hippocampus kuda bleeler. Here, we examined the antitumor metastatic activity of this peptide and investigated its mechanism. The peptide significantly inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced invasive migration of human osteosarcoma MG-63 cells. Its inhibitory effect on invasive migration was associated with reduced expression of matrix metalloproteinases (MMP1 and MMP2). In addition, TPA stimulation increased intracellular reactive oxygen species (ROS) generation and small GTPase Rac1 expression, whereas the peptide decreased ROS generation and Rac1 activation. Taken together, these results suggest that the peptide inhibits invasive migration of MG-63 osteosarcoma cells by inhibiting MMP1 and MMP2 expression through downregulation of Rac1-ROS signaling.

• International Journal of Molecular Medicine
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• v.44 no.5
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• pp.1979-1987
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• 2019

Sargassum thunbergii is a brown alga from which various bioactive compounds can be extracted. Among these, the activities of indole derivatives, particularly as potential inhibitors of matrix metalloproteinases (MMPs), and their underlying mechanisms have been rarely investigated. Therefore, we evaluated the inhibitory effects of indole-6-carboxaldehyde (I6CA) on MMP-9 by gelatin zymography and western blot anlaysis. We used phorbol 12-myristate 13-acetate (PMA), which is known to induce MMP-9 expression and secretion, to stimulate HT1080 cells. Our results revealed that I6CA significantly inhibited MMP-9 expression and secretion, without significantly affecting the viability of PMA-stimulated HT1080 cells. Our mechanistic studies indicated that I6CA suppressed the phosphorylation and activation of two mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (ERK). Furthermore, I6CA inhibited the phosphorylation of inhibitor of κBα (IκBα) in response to PMA stimulation, which suppressed nuclear factor-κB (NF-κB) p65 subunit nuclear translocation. Collectively, I6CA was determined to suppress MMP-9 expression and secretion, and effects were proposed to be mediated via the inhibition of the MAPK and NF-κB p65 pathways. Therefore, we suggested I6CA to be a potential therapeutic agent for MMP-9-related processes, including tumor invasion and metastasis; however, further investigation is required.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.3
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• pp.611-621
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• 2016

This study examined the effects of collagen peptide intake extracted from the tilapia fish (TFCP) on UVB-induced skin in hairless mice. To cause photoaging, the hairless mice were exposed to UVB irradiation three times a week and TFCP was administered with 545 and 1090 mg/kg once a day for 12 weeks. The level of wrinkle formation, thickness of the skin, acute erythema, skin hydration, amount of hydroxyproline, expression of MMPs and filaggrin were measured. Wrinkle formation, thickness of the skin, and acute erythema were significantly lower and skin hydration was higher in the TFCP intake group compared to the UVB-control mice group. In addition, the amount of the collagen and hydroxyproline of the collagen peptide intake group were much higher than the UVB-control group. Oral administration of TFCP inhibited the expression of the MMP-3 and MMP-13 mRNA level as well as activated MMP-2 and MMP-9 induced by UV irradiation, and increased the protein expression of filaggrin in hairless mice skin. In summary, the oral administration of TFCP can inhibit wrinkle formation and protect the skin from UV-damage. Therefore, this study suggests that TFCP is a potential candidate material for skin functional foods.

• The Korea Journal of Herbology
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• v.37 no.6
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• pp.37-44
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• 2022

Objectives : From this study, we sight to identify chondro-protective and anti-inflammatory effects of Sorbi Commixtae Fructus extract and its compound, chlorogenic acid. Methods : Sorbi Commixtae Fructus were extracted by 50% ethanol. And chlorogenic acid in Sorbi Commixtae Fructus 50% extract was quantified by high performance liquid chromatography (HPLC). To investigate chondro-protective effects, we treated Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid in TNF𝛼-activated ATDC5 murine chondrogenic cells. After 24 hours, protein level of matrix metalloproteinase-3 (MMP3) and mRNA level of matrix metalloproteinase-13 (MMP13) were measured by using ELISA or reverse transcription PCR, respectively. To examine anti-inflammatory effects, we treated Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid in LPS-induced RAW 264.7 murine macrophages. We measured the level of inflammatory mediators, such as Prostaglandin E2 (PGE2), Interleukin-6 (IL6) by ELISA and nitric oxide (NO) by Griess reagent assay. Results : A concentration of chlorogenic acid in Sorbi Commixtae Fructus 50% ethanol extract was 3.9 mg/g. Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid attenuated protein level of MMP3 and mRNA level of MMP13 in TNF𝛼-activated ATDC5 cells. Sorbi commixtae Fructus 50% ethanol extract inhibited the level of PGE2, IL6 and NO in LPS-activated RAW 264.7 cells in dose dependent manner, but chlorogenic acid has no anti-inflammatory effects. Conclusions : These findings demonstrated that Sorbi Commixtae Fructus 50% ethanol extract has chondro-protective and anti-inflammatory effects showing possible therapeutics to ease the symptoms related with osteoarthritis.

• The Korea Journal of Herbology
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• v.37 no.6
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• pp.1-8
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• 2022

Objectives : This research aimed to investigate chondro-protective and anti-inflammatory effects of Caraganae Sinicae Flos 50% ethanol extract and its compound, tilianin. Methods : Caraganae Sinicae Flos was extracted with 50% ethanol. Tilianin in Caraganae Sinicae Flos 50% ethanol extract was quantified by HPLC analysis method. To investigate chondro-protective effects of Caraganae Sinicae Flos 50% ethanol extract, ATDC5 chondrogenic cells were co-treated with Caraganae Sinicae Flos 50% ethanol extract (or tilianin) and tumor necrosis factor-𝛼 (TNF𝛼) for 24 hours. After treatement for 24 hours, media supernatant was used for quantifying protein level of matrix metalloproteinase-3 (MMP3) by ELISA and harvested cells were used for analyzing mRNA expression level of matrix metalloproteinase-13 (MMP13) by reverse transcription PCR. To identify anti-inflammatory effects of Caraganae Sinicae Flos 50% ethanol extract, RAW 264.7 macrophage cells were co-treated with Caraganae Sinicae Flos 50% ethanol extract (or tilianin) and lipopolysaccharide (LPS) for 24 hours. media was used for quantifying the level of prostaglandin E2 (PGE2), interleukin-6 (IL6) by ELISA and nitric oxide by Griess reagent asssay. Results : Caraganae Sinicae Flos 50% ethanol extract and tilianin attenuated protein level of MMP3 and mRNA expression level of MMP13 in TNF𝛼-activated ATDC5 cells. Caraganae Sinicae Flos 50% ethanol extract inhibited the level of PGE2, IL6 and NO in LPS-activated RAW 264.7 cells in dose dependent manner, though tilianin inhibited PGE2 only. Conclusions : These results presented that Caraganae Sinicae Flos 50% ethanol extract could be used as natural medicines for osteoarthritis.

• Tuberculosis and Respiratory Diseases
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• v.56 no.1
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• pp.51-66
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• 2004

Objectives : The matrix metalloproteinases (MMPs) that participate in the extracellular matrix metabolism play a important role in the progression of pulmonary fibrosis. The effects of the MMPs are regulated by several factors including Th-1 cytokines, $interferon-{\gamma}$ ($IFN-{\gamma}$). Up to now, $IFN-{\gamma}$ is known to inhibit pulmonary fibrosis, but little is known regarding the exact effect of $IFN-{\gamma}$ on the regulation of the MMPs. This study investigated the effects of $interferon-{\gamma}$ on the pulmonary fibrosis and the expression of the lung MMP-2,-9, TIMP-1,-2, and Th-2 cytokines in aa rat model of bleomycin induced pulmonary fibrosis. Materials and methods : Male, specific pathogen-free Sprague-Dawley rats were subjected to an intratracheal bleomycin instillation. The rats were randomized to a saline control, a bleomycin treated, and a bleomycin+$IFN-{\gamma}$ treated group. The bleomycin+$IFN-{\gamma}$ treated group was subjected to an intramuscular injection of $IFN-{\gamma}$ for 14 days. At 3, 7, 14, and 28 days after the bleomycin instillation, the rats were sacrificed and the lungs were harvested. In order to evaluate the effects of the $IFN-{\gamma}$ on lung fibrosis and inflammation, the lung hydroxyproline content, inflammation and fibrosis score were measured. Western blotting, zymography and reverse zymography were performed at 3, 7, 14, 28 days after bleomycin instillation in order to evaluate the MMP-2,-9, and TIMP-1,-2 expression level. ELISA was performed to determine the IL-4 and IL-13 level in a lung homogenate. Results : 1. 7 days after bleomycin instillation, inflammatory changes were more severe in the bleomycin+$IFN-{\gamma}$ group than the bleomycin group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$2.08{\pm}0.15:2.74{\pm}0.29$, P<0.05), but 28 days after bleomycin instillation, lung fibrosis was significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$3.94{\pm}0.43:2.64{\pm}0.13$, P<0.05). 2. 28 days after bleomycin instillation, the lung hydroxyproline content was significantly reduced as a result of $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$294.04{\pm}31.73{\mu}g/g:194.92{\pm}15.51{\mu}g/g$, P<0.05). 3. Western blotting showed that the MMP-2 level was increased as a result of the bleomycin instillation and highest in the 14 days after bleomycin instillation. 4. In zymography, the active forms of MMP-2 were significantly increased as a result of the $IFN-{\gamma}$ treatment 3 days after the bleomycin instillation, bleomycin+$IFN-{\gamma}$ group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$209.63{\pm}7.60%:407.66{\pm}85.34%$, P<0.05), but 14 days after the bleomycin instillation, the active forms of MMP-2 were significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$159.36{\pm}20.93%:97.23{\pm}12.50%$, P<0.05). 5. The IL-4 levels were lower in the bleomycin and bleomycin+$IFN-{\gamma}$ groups but this was not significant, and the IL-13 levels showed no difference between the experiment groups. Conclusion : The author found that lung inflammation was increased in the early period but the pulmonary fibrosis was inhibited in the late stage as a result of $IFN-{\gamma}$. The inhibition of pulmonary fibrosis by $IFN-{\gamma}$ appeared to be associated with the inhibition of MMP-2 activation by $IFN-{\gamma}$. Further studies on the mechanism of the regulation of MMP-2 activation and the effects of MMP-2 activation on pulmonary fibrosis is warranted in the future.

• Clinics in Shoulder and Elbow
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• v.25 no.4
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• pp.296-303
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• 2022

Background: A previous study reported that hyperlipidemia increases the incidence of tears in the rotator cuff tendon and affects healing after repair. The aim of our study was to compare the gene and protein expression of torn rotator cuff tendons in patients both with and without hypercholesterolemia. Methods: Thirty patients who provided rotator cuff tendon samples were classified into either a non-hypercholesterolemia group (n=19, serum total cholesterol [TC] <200 mg/dL) and hypercholesterolemia group (n=11, serum TC ≥240 mg/dL) based on their concentrations of serum TC. The expression of various genes of interest, including COL1A1, IGF1, IL-6, MMP2, MMP3, MMP9, MMP13, TNMD, and TP53, was analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, Western blot analysis was performed on the proteins encoded by interleukin (IL)-6 and TP53 that showed significantly different expression levels in real-time qRT-PCR. Results: Except for IGF1, the gene expression levels of IL-6, MMP2, MMP9, and TP53 were significantly higher in the hypercholesterolemic group than in the non-hypercholesterolemia group. Western blot analysis confirmed significantly higher protein levels of IL-6 and TP53 in the hypercholesterolemic group (p<0.05). Conclusions: We observed an increase in inflammatory cytokine and matrix metalloproteinase (MMP) levels in hypercholesterolemic patients with rotator cuff tears. Increased levels of IL-6 and TP53 were observed at both the mRNA and protein levels. We suggest that the overexpression of IL-6 and TP53 may be a specific feature in rotator cuff disease patients with hypercholesterolemia.

• Journal of Ginseng Research
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• v.39 no.1
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• pp.54-60
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• 2015

Background: The present study was designed to prepare and find the optimum active preparation or fraction from Korea Red Ginseng inhibiting matrix metalloproteinase-13 (MMP-13) expression, because MMP-13 is a pivotal enzyme to degrade the collagen matrix of the joint cartilage. Methods: From total red ginseng ethanol extract, n-BuOH fraction (total ginsenoside-enriched fraction), ginsenoside diol-type-enriched fraction (GDF), and ginsenoside triol-type-enriched fraction (GTF) were prepared, and ginsenoside diol type-/F4-enriched fraction (GDF/F4) was obtained from Panax ginseng leaf extract. Results: The n-BuOH fraction, GDF, and GDF/F4 clearly inhibited MMP-13 expression compared to interleukin-$1{\beta}$-treated SW1353 cells (human chondrosarcoma), whereas the total extract and ginsenoside diol-type-enriched fraction did not. In particular, GDF/F4, the most effective inhibitor, blocked the activation of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun-activated protein kinase (JNK), and signal transducer and activator of transcription-1/2 (STAT-1/2) among the signal transcription pathways involved. Further, GDF/F4 also inhibited the glycosaminoglycan release from interleukin-$1{\alpha}$-treated rabbit cartilage culture (30.6% inhibition at $30{\mu}g/mL$). Conclusion: Some preparations from Korean Red Ginseng and ginseng leaves, particularly GDF/F4, may possess the protective activity against cartilage degradation in joint disorders, and may have potential as new therapeutic agents.

• Analytical Science and Technology
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• v.18 no.2
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• pp.104-111
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• 2005

Several solvents were used to fractionate an extract obtained from the chapped root bark of Ulmus davidiana Planchon. The each fractionary part was condensed under reduced pressure and then examined to investigate the inhibitory effect on MMPs by modified gelatin zymography, where EA fraction showed the inhibition effect on the activity of MMPs. A compound showing inhibition effect on the MMPs was isolated and purified from EA fraction. Under IR, $^1H$- and $^{13}C$- NMR analyses it is very close to a catethin. This substance showed 48% inhibition effect on measurement of MMP-9 activity at 5 mM and 43% at 10 mM. To verify the effect of this substance on cells, human hepatoma, SK-Hep-1 cells as a cancer model, and Chang liver cells as a normal model were selected. MTT assay was performed to examine the cell viability by treatment of $1{\mu}L/mL$ of the purified substance on cells. The purified substance showed negligible toxicity on human liver cell line.

• CELLMED
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• v.8 no.3
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• pp.13.1-13.8
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• 2018

In the field of osteoporosis, there has been growing interest in anabolic agents that enhance bone formation. The purpose of this study was to examine the effects of NNMBS 246 osteoblastic differentiation with associated signaling pathways. NNMBS 246 markedly increased alkaline phosphatase (ALP) activity and calcium nodule formation. Stimulation with NNMBS 246 not only increased the differentiation markers (ALP, OPN, OCN) level and transcription markers (RUNX2, Osterix) mRNA expression but also upregulated the ECM molecules and OPG mRNA expression. Treatments of NNMBS 246 downregulated MMPs (MMP-1, MMP-2, MMP-9), but RANKL mRNA expression. Furthermore, NNMBS 246 activated osteoblastic differentiation markers and formed calcium nodules in human periodontal ligament cells (hPDLCs) and cementoblast cells. NNMBS 246 induced phosphorylation of MAPKs, Akt, nuclear p65 and IkB-${\alpha}$. BMP-2/Smad and ${\beta}$-catenin signaling pathways were activated by NNMBS 246. Sirtinol (SIRT1 inhibitor) inhibited NNMBS 246-induced osteoblastic differentiation markers mRNA expression. These results suggested that NNMBS 246 has the potential to enhance osteoblastogenesis probably through the activation of BMP/Smad and ${\beta}$-catenin signal pathways, and SIRT1 plays as critical mediator in bone anabolic effect of NNMBS 246.