• Title/Summary/Keyword: MMPs

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Proteinases and their Inhibitors in Cartilage and Synovial Fluid Acquired from a Canine Osteoarthritic Model (개 퇴행성 관절염 모델을 이용한 연골과 활액 내 단백질 분해 효소와 억제제의 작용 연구)

  • Seo, Jae-Won;Lee, Hae-Beom;Kim, Nam-Soo;Lee, Young-Hoon;Kang, Hyung-Sub;Kim, In-Shik;Park, Sang-Youel
    • Journal of Veterinary Clinics
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    • v.26 no.2
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    • pp.144-149
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    • 2009
  • Chondrocytes and synovial fluid derived markers are used to monitor for osteoarthritis(OA). Specific inhibitors, known as tissue inhibitors of metalloproteinases(TIMP), regulate the proteolytic activity of matrix metalloproteinases(MMP). This study investigated whether MMP and TIMP levels were altered in synovial fluid and cartilage following the experimental induction of OA in canines. Twenty mature beagle dogs underwent a unilateral surgical transection of the cranial cruciate ligament and the medial collateral ligament as well as a medial meniscectomy. Matrix metalloproteinase-2 and MMP-9 levels were assayed using Western blot and TIMP-2 levels were measured with enzyme-linked immunosorbent assays four weeks after OA induction. Increased MMP-2 expression was observed in chondrocytes isolated from cartilage following OA induction, but MMP-9 expression decreased. Matrix metalloproteinase-2 and MMP-9 levels in synovial fluid from the OA induced joint significantly increased compared to those of the sham group. Tissue inhibitors of metalloproteinase-2 concentrations were higher in chondrocytes from the OA cartilage, yet TIMP-2 remained lower in the synovial fluid of OA. This suggests the elevated release of MMP-9 over MMP-2 into the synovial fluid following the cartilage degradation-related death of chondrocytes after OA. Osteoarthritis can be further deteriorated by increased MMP activity in the synovial fluid because TIMP-2 exist low concentration into the extracellular matrix. As a result, MMP activity, particularly MMP-9 activity, can be useful as a biomarker in diagnosing and monitoring the early stages of canine OA.

The Anti-angiogenic Potential of a Phellodendron amurense Hot Water Extract in Vitro and ex Vivo (in Vitro와 ex vivo에서 황백 온수추출물의 신생혈관 억제효과)

  • Kim, Eok-Cheon;Kim, Seo Ho;Bae, Kiho;Kim, Han Sung;Gelinsky, Michael;Kim, Tack-Joong
    • Journal of Life Science
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    • v.25 no.6
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    • pp.693-702
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    • 2015
  • Blocking new blood-vessel formation (angiogenesis) is now recognized as a useful approach to the therapeutic treatment of many solid tumors. The best validated approach to date is to target the vascular endothelial growth-factor (VEGF) pathway, a key regulator of angiogenesis. Many natural products and extracts that contain a variety of chemopreventive compounds have been shown to suppress the development of malignancies through their anti-angiogenic properties. Phellodendron amurense, which is widely used in Korean traditional medicine, has been shown to possess antitumor, antimicrobial, and anti-inflammatory properties, among others. The present study investigated the effects of P. amurense hot-water extract (PAHWE) on angiogenesis, a key process in tumor growth, invasion, and metastasis. To investigate PAHWE’s anti-angiogenic properties, this study’s authors performed an analysis of angiogenesis and endothelial-cell proliferation, migration, invasion, and tube formation, as well as zymogram assays and the rat aortic ring-sprouting assay. PAHWE inhibited cell growth, mobility, and vessel formation in response to VEGF in vitro and ex vivo. Furthermore, it reduced VEGF-induced intracellular signaling events, such as the activation of matrix metalloproteinases (MMPs) -2 and -9. These results indicate that PAHWE’s anti-angiogenic properties might lead to the development of potential drugs for treating angiogenesis-associated diseases such as cancer.

Anti-aging Effects of L-Carnitine on Human Skin (L-카르니틴의 사람피부에 대한 항노화 효과)

  • Lee Bum-Chun;Choe Tae-Boo;Sim Gwan-Sub;Lee Geun-Soo;Park Sung-Min;Lee Chun-Il;Pyo Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.393-397
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    • 2004
  • L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

Antioxidant and Antiwrinkle Effects of Persimmon Leaves extract (시엽(Persimmon Leaves) 에탄올 추출물의 항산화와 항주름 효과)

  • Sung-Hee Kim;Dong-Hee Kim;Wi-Hye Yeon;Jin-Tae Lee;Young-Ah Jang
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.3
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    • pp.534-546
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    • 2023
  • In this study, we investigated the antioxidant and anti-winkle activity in human fibroblast cell (CCD-986sk) of Persimmon Leaves (PL) as a cosmetic ingredient. As a result of investigating antioxidant activity through electron-donating ability and ABTS+ radical scavenging assay, the PL showed concentration-dependent antioxidant activity similar to ascorbic acid, a control group, at a concentration of 1,000 ㎍/ml. As a result of investigating the anti-wrinkle effect through elastase inhibition and collagenase inhibition assay, the PL showed concentration-dependent antioxidant activity similar to epigallocatechin gallate, a control group, at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I and the inhibition rate of MMP-1 in UVB-induced CCD-986sk cells, the control group EGCG showed a 90.2% pro-collagen synthesis rate at 20 ㎍/ml and PL showed an 88.5% synthesis rate at 30 ㎍/ml. In addition, the inhibition rate of MMP-1 of 33.0% and 40.8% were confirmed in EGCG 20 ㎍/ml and PL 30 ㎍/ml, respectively. As a result of measuring the protein expression of pro-collagen type I and MMP-1 in the PL through western blot, it was confirmed that the protein expression of pro-collagen type I increased, and MMP-1 decreased when the PL was treated together compared to the UVB alone group. According to the above experimental results, it is expected to be used as a natural product material for cosmetics by confirming that the PL prevent photoaging caused by UVB stimulation and have antioxidant and anti-wrinkle effects.

Evaluation of Anti-wrinkle Effect of Peanut Shell Extract and Stability of the Extract in Cosmetic Products (땅콩 겉껍질 추출물의 주름개선 효능 및 화장품 제형에서 추출물의 안정성 평가)

  • Narae Han;Jin Young Lee;Mihyang Kim;Eun Young Choi;Bong-Jeun An;Yu-Young Lee;Moon Seok Kang;Hyun-Joo Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.203-212
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    • 2023
  • This study was conducted to investigate the possibility of peanut shell, a by-product of peanut, as a functional cosmetic ingredient. Peanut shell extract showed high antioxidant activity with IC50 values of 75.00, 46.33, and 472.83 ㎍/mL for DPPH and ABTS radical scavenging and SOD-like activity, respectively. Furthermore, peanut shell extract was efficiently decreased the MMP-1 and MMP-3 protein level in the UVB treated-HaCaT cell and maintained procollagen protein level similar to normal control. Similar to anti-wrinkle related protein expression assay, the IC50 value of elastase and collagnease inhibition in peanut shell extract was lower as 0.30 and 0.09 mg/mL, respectively, than that of the positive control. Additionally, eriodictyol and luteolin, which are isolated from peanut shell extract, showed 53.8 and 98.0% elastase inhibition rate, respectively, and 60.1 and 72.5% collagenase inhibition rate, respectively, at a concentration of 0.1 mg/mL. Thus, luteolin was assumed to be the effective ingredient for wrinkle inhibition in peanut shell extract. As a result of stability evaluation of lotion and cream formulations containing peanut shell extract, it was confirmed to be a stable formulation with no significant changes. Therefore, it is considered that peanut shell extract can be applied as a cosmetic ingredient for wrinkle inhibition.

Anti-wrinkle effects of solvent fractions from Jubak on CCD-986sk (CCD-986sk 세포 내 주박 분획물의 항주름 효능)

  • Young-Ah Jang;Hyejeong Lee
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.2
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    • pp.508-519
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    • 2024
  • In this study, in order to evaluate the possibility of using Jubak as a functional cosmetic material, evaluation of antioxidant activity according to fractions and anti-wrinkle efficacy in CCD-986sk cells, a human fibroblast, were conducted. As a result of confirming the antioxidant activity by measuring ABTS+ radical scavenging ability, Jubak's Ethyl Acetate fractions was found to be 75.5% at a concentration of 1,000 ㎍/ml, showing the highest antioxidant activity among the extraction solvents. The wrinkle improvement effect was confirmed by measuring the inhibitory activity of elastase and collagenase, and in both test results, Jubak's Ethyl Acetate fractions showed the highest efficacy at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I in CCD-986sk cells induced by UVB, Jubak showed the highest efficacy in the order of Ethyl Acetate, Water, Acetonitrile, and Hexan fractions at the same concentration of 20 ㎍/ml. As a result of measuring the inhibition rate of MMP-1, a collagen degrading enzyme, all four solvent fractions showed an efficacy of more than 70% at 20 ㎍/ml. As a result of measuring the mRNA expression levels of pro-collagen type I, MMP-1, and MMP-3 in a real-time PCR experiment, the protein expression level of pro-collagen type I increased when treated with Jubak fractions compared to the UVB group alone. The mRNA expression levels of MMP-1 and MMP-3 were confirmed to be decreased, and Ethyl Acetate fractions was the most effective in improving wrinkles after the control group (EGCG). As a result, it was confirmed that the Ethyl Acetate fractions among Jubak's solvent fractions has an anti-wrinkle effect against photoaging caused by UVB stimulation, and is expected to be used as a natural material for cosmetics.