• Journal of Applied Biological Chemistry
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• 제62권1호
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• pp.101-107
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• 2019

The objectives of this study were to evaluate the anti-aging effects and investigate the effect of simulated gastrointestinal (GI) digestion on the anti-aging properties and intestinal permeation of the potential fish collagen hydrolysates (FCH). Therefore, procollagen synthesis, matrix metalloproteinase-1 (MMP-1) production, and Caco-2 cell permeability were analyzed before and after in vitro digestion for FCHs, low-molecular weight fractions (<1 kDa), and high molecular weight fractions (>1 kDa). After being subjected to GI digestion, the level of MMP-1 inhibition was maintained, although the procollagen production was significantly (>20%) lower with all samples. Also, the digested FCHs and their <1 kDa fraction yielded 9.1 and 13.8% increased peptide transport, respectively, compared to undigested samples. Based on the effective intestinal permeation and high digestive enzyme stability, the <1 kDa fraction of FCHs is a potential bioactive material suitable for anti-aging applications in the food and cosmetics industries.

• Radiation Oncology Journal
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• 제21권1호
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• pp.66-74
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• 2003

목적 : 금속단백효소(MMPs)는 세포외 기질의 분해가 주 기능이며 금속단백효소와 억제자(TIMPS)가 상처 치유 과정과 자외선 조사에 의한 피부노화에 관여한다는 것이 알려져 있다. 그러나 방사선조사 후 금속단백효소와 억제자의 발현에 관한 연구는 보고된 바 없다. 본 연구는 흰쥐의 소장에 방사선을 조사하여 금속단백효소와 억제자의 발현을 시간적으로 관찰하여 방사선에 의한 소장 점막 손상 및 재생과정과 금속단백효소와 억제자의 관계를 밝히고자하였다. 대상 및 방법 : 실험동물로 암, 수 구별 없이 생후 4~5개월, 체중 250~300 gm의 흰쥐(Spraque-Dawley) 30마리를 대상으로 하여 실험군으로 전복부에 8 Gy의 방사선을 조사한 후 1일, 2일, 3일, 5일, 7일, 14일에 각각 5마리씩 희생시켜 소장을 적출하여 사용하였고, 정상대조군은 각 시기별로 1마리씩 사용하였다. H&E 염색을 하여 조직병리검사를 하였고, MMP와 TIMP끌의 발현에 대해서는 면역조직화학염색, 면역블로팅, ELISA법으로 확인하였다. 결과 : 조직병리 소견상 방사선조사에 의한 소장 조직손상은 1일부터 관찰되어 2일과 3일에 현저하였고 재생은 3일에 관찰되어 5일에 현저하였다 면역조직화학염색 결과 MMP는 1일에 발현되어 3일, 5일에 가장 강하게 나타났으며 TIMP를는 1일에 나타나기 시작하여 5일에 가장 강하게 나타났다. 면역블로팅 결과 MMP◎는 3일과 5일에강하게 나타났으며 TIMP끌는 1, 3, 5, 7일에 같은 정도의 양성반응을 보였다. 혈액에서의 MMP와 TIMP의 ELISA 검사법에서는 대조군에 비해 방사선조사군에서 통계적으로 유의한 증가가 있었고 방사선조사 후 1, 2, 3, 5, 7, 14일 각각에서 유의한 차이가 있었으며 5일에 가장 높은 수치를 보였다. 결론 : 방사선조사 후 조직병리학적 검사에서 손상의 척도인 염증 반응과 재생의 척도인 유사분열 수의 변화가 면역조직화학염색, 면역블로팅, ELISA검사에 의한 MMP-2, TIMP의 증가와 일치하는 것으로 나타나 이들이 방사선 조사 후 조직 손상과 재생 기전에 역할을 하고 있음을 보여주었다.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.590-600
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• 2003

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.68.3-69
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• 2003

Matrix metalloproteinases (MMPs) play an important role in tumor invasion and metastasis by extracellular matrix degradation. To analyze the effect of DA-125, a anthracyclin derivative, on the invasion or metastasis of cancer cells the expression of matrix metalloproteases (MMPs) was investigated in human fibrosarcoma HTl080 cells by RT-PCR or gelatin zymographic methods. As result, DA-125 suppressed the expression of MMP-2 and 9 as well as tissue inhibitor of metalloproteinase-1 (TIMP-1) TIMP-2 and MT1-MMP with a time- and dose-dependent manner. Inaddition, DA-125 inhibited cancer cell migration and colony formation, and also exhibited the inhibitory activities of invasion and motility with a matrigel and type I collagen assay. (omitted)

• Journal of Ginseng Research
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• 제45권1호
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• pp.119-125
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• 2021

Background: Korean Red Ginseng (KRG) is a natural product with antiinflammatory and anticarcinogenic effects. We have previously reported that the endocrine-disrupting compound bisphenol A (BPA)-induced cyclooxygenase-2 (COX-2) via nuclear translocation of nuclear factor-kappa B (NF-κB) and activation of mitogen-activated protein kinase and promoted the migration of A549. Here, in this study, we assessed the protective effect of KRG on the BPA-induced reactive oxygen species (ROS) and expression of COX-2 and matrix metalloproteinase-9 (MMP-9) in A549 cells. Methods: The effects of KRG on the upregulation of ROS production and COX-2 and MMP-9 expression by BPA were evaluated by fluorescence-activated cell sorting (FACs) analysis, quantitative reverse transcription polymerase chain reaction, and western blotting. Antimigration ability by KRG was evaluated by migration assay in A549 cells. Results: KRG significantly suppressed the BPA-induced COX-2, the activity of NF-κB, the production of ROS, and the migration of A549 cells. These effects led to the downregulation of the expression of MMP-9. Conclusions: Overall, our results suggest that KRG exerts an antiinflammatory effect on BPA-treated A549 cells via the suppression of ROS and downregulation of NF-κB activation and COX-2 expression which leads to a decrease in cellular migration and MMP-9 expression. These results provide a new possible therapeutic application of KRG to protect BPA-induced possible inflammatory disorders.

• 대한본초학회지
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• 제37권6호
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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.

• 대한본초학회지
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• 제37권6호
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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.

• The Korean Journal of Pain
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• 제35권3호
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• pp.291-302
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• 2022

Background: Spinal cord injury (SCI) is one of the most debilitating disorders throughout the world, causing persistent sensory-motor dysfunction, with no effective treatment. Oxidative stress and inflammatory responses play key roles in the secondary phase of SCI. Naringenin (NAR) is a natural flavonoid with known anti-inflammatory and antioxidative properties. This study aims at evaluating the effects of intrathecal NAR administration on sensory-motor disability after SCI. Methods: Animals underwent a severe compression injury using an aneurysm clip. About 30 minutes after surgery, NAR was injected intrathecally at the doses of 5, 10, and 15 mM in 20 µL volumes. For the assessment of neuropathic pain and locomotor function, acetone drop, hot plate, inclined plane, and Basso, Beattie, Bresnahan tests were carried out weekly till day 28 post-SCI. Effects of NAR on matrix metalloproteinase (MMP)-2 and MMP-9 activity was appraised by gelatin zymography. Also, histopathological analyses and serum levels of glutathione (GSH), catalase and nitrite were measured in different groups. Results: NAR reduced neuropathic pain, improved locomotor function, and also attenuated SCI-induced weight loss weekly till day 28 post-SCI. Zymography analysis showed that NAR suppressed MMP-9 activity, whereas it increased that of MMP-2, indicating its anti-neuroinflammatory effects. Also, intrathecal NAR modified oxidative stress related markers GSH, catalase, and nitrite levels. Besides, the neuroprotective effect of NAR was corroborated through increased survival of sensory and motor neurons after SCI. Conclusions: These results suggest intrathecal NAR as a promising candidate for medical therapeutics for SCI-induced sensory and motor dysfunction.

• 대한본초학회지
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• 제38권4호
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• pp.45-52
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• 2023

Objectives : Fine dust caused by environmental pollution cause oxidative damage and skin aging. In this study, The possibility of using the Codium fragile extract (CFE) as an anti-aging product for skin improvement was evaluated by confirming the protective effect of skin cells from PM10 (particulate matter 10) through inhibition of ROS and MMPs. Methods : In this study, elastase and collagenase inhibitory activities were evaluated. Cell viability was evaluated by treating keratinocytes (HaCaT cell line) with CFE at various concentrations. The cytoprotective effect from PM10 in keratinocyteswas evaluated using the 3-[4,5-dimethylthiazol]-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay. ROS (reactive oxygen species) was measured in keratinocytes damaged by PM10 using DCF-DA (2′,7′-dichlorofluorescin diacetate) fluorescence staining. As an anti-aging effect of CFE, MMP-1 (matrix metalloproteinase-1) and MMP-1 (matrix metalloproteinase-9) inhibitory activities were evaluated. Results : As a result, CFE decreased the activity of elastase and collagenase. As a result of evaluating the toxicity of CFE, it is non-toxic at a concentration of 10 to 80 ㎍/㎖. Although cell viability of HaCaT cells treated with PM10 decreased, cell viability increased by 38% when treated with CFE 80 ㎍/㎖. Also, ROS decreased by 8.4%, and MMP-1 and MMP-9 decreased at CFE 80 ㎍/㎖. Conclusions : CFE showed excellent cell protection effect, and it is considered that it can be used in anti-aging products for skin improvement by effectively inhibiting ROS and MMPs from keratinocyte damage caused by fine dust.

• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.334-342
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• 2013

Proteases and protease inhibitors play key roles in most physiological processes, including cell migration, cell signaling, and cell surface and tissue remodeling. Among these, the matrix metalloproteinase (MMPs) pathway is one of the most efficient biosynthetic pathways for controlling the activation of enzymes responsible for protein degradation. This also indicates the association of MMPs with the maturation of spermatozoa. In an attempt to investigate the effect of MMP activation and inhibitors in cultures with various hormones during sperm capacitation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3), as well as their expression profiles. Matured spermatozoa were collected from cultures with follicle-stimulating hormone (FSH), luteinizing hormone (LH), and Lutalyse at 1 h, 6 h, 18 h, and 24 h. ELISA detected the expression of MMP-2, MMP-9, TIMP-2, and TIMP-3 in all culture media, regardless of medium type (FSH-supplemented fertilization Brackett-Oliphant medium (FFBO), LH-supplemented FBO (LFBO), or Lutalyse-supplemented FBO (LuFBO)). TIMP-2 and TIMP-3 expression patterns decreased in LFBO and LuFBO. MMP-2 and MMP-9 activity in FBO and FFBO progressively increased from 1 h to 24 h but was not detected in LFBO and LuFBO. The localization and expression of TIMP-2 and TIMP-3 in sperm heads was also measured by immunofluorescence analysis. However, MMPs were not detected in the sperm heads. MMP and TIMP expression patterns differed according to the effect of various hormones. These findings suggest that MMPs have a role in sperm viability during capacitation. In conjunction with hormones, MMPs play a role in maintaining capacitation and fertilization by controlling extracellular matrix inhibitors of sperm.