• Title/Summary/Keyword: collagenase activity

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Effect of Cervi Pantotrichum Cornu Herbal acupuncture on protease activities, antioxidant in Rheumatoid arthritis rats (류마티스 관절염 실험용쥐의 활액에서 단백분해효소의 활성 및 항산화에 대한 녹용약침의 효과)

  • Park, Sang-Dong;Kim, Min-Jeong;Lee, A-Ram;Jang, Jun-Hyouk;Kim, Kyung-Ho
    • Journal of Acupuncture Research
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    • v.19 no.2
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    • pp.51-64
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    • 2002
  • We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

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Biological Activities of Extracts from Okkwang (Castanea crenata) Chestnut Bur (옥광(Castanea crenata) 밤송이 추출물의 생리활성 효과)

  • Lee, Eun Ho;Hong, Shin Hyub;Cho, Young Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.572-580
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    • 2017
  • The contents of phenolic compounds in water and 40% ethanol extracts from Okkwang (Castanea crenata) chestnut bur solid (OCS) were $11.24{\mu}g/50{\mu}g$ solid and $10.28{\mu}g/50{\mu}g$ solid, respectively. The 1,1-diphenyl-2-picrylhydrazyl free radical scavenging and 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulfonic acid) radical decolorization activities of water and ethanol extracts were 85% and 100% as well as 87% and 86% at a solid content of $50{\mu}g/mL$, respectively. The anti-oxidant protection factors (PFs) of water and ethanol extracts at a solid content of $200{\mu}g/mL$ were 1.22 PF and 1.45 PF, respectively. Thiobarbituric acid reactive substance were 83% in water extract and 73% in ethanol extract at a solid content of $200{\mu}g/mL$. The inhibitory activities against xanthine oxidase in water and ethanol extracts were 54% and 43% at a solid content of $200{\mu}g/mL$, respectively. The inhibitory activities against ${\alpha}$-glucosidase were 95% in water extract and 96% in ethanol extract at a solid content of $50{\mu}g/mL$. Tyrosinase inhibitory activity was 27% in ethanol extract at a solid content of $200{\mu}g/mL$. The collagenase and elastase inhibitory activities as anti-wrinkle effect were 93% and 11% in water extract as well as 94% and 56% in ethanol extract at a solid content of $200{\mu}g/mL$. Hyaluronidase inhibitory activity as anti-inflammatory effect of water and ethanol extracts were 96% and 52% at a solid content of $200{\mu}g/mL$, respectively. The results show that extracts from OCS can be used as a functional resource with antioxidant, anti-gout, carbohydrate degradation inhibitory, whitening, anti-wrinkle, and anti-inflammatory activities.

Protective Effects of Portulaca oleracea L. Extract against Matrix Metalloproteinase Production and Reactive Oxygen Species Generation Induced by Ultraviolet B Radiation in Human Keratinocytes (쇠비름 추출물의 UVB 자외선 조사에 의한 인간각질형성세포 손상에 대한 보호 효과)

  • Oh, Jung Hwan;Karadeniz, Fatih;Lee, Jung Im;Park, So Young;Seo, Youngwan;Kong, Chang-Suk
    • Journal of Life Science
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    • v.28 no.8
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    • pp.892-899
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    • 2018
  • Portulaca oleracea L. is an edible plant widely consumed in daily diet throughout Europe, Asia and America. In this study, protective effects of P. oleracea L. extracts against oxidative stress and matrix metalloproteinase (MMP) activity induced by ultraviolet B (UVB) radiation were investigated using HaCaT immortal human keratinocytes. In this context, the mRNA and protein productions of MMPs (MMP-1, -2, and -9) and type I procollagen, which are major markers of photoaging induced by UVB radiation in HaCaT keratinocytes, were evaluated. Furthermore, UVB-induced reactive oxygen species (ROS) generation and mRNA and protein expression levels of superoxide dismutase-1 (SOD-1), oxygenase-1 (OH-1), and nuclear factor-erythroid 2-related factor-2 (Nrf-2), all of which are associated with the antioxidant balance, were investigated. As shown by the results, UVB radiation induced ROS formation and led to increased production of MMPs and decreased collagen production in human keratinocytes, which resulted in skin photoaging or photodamage. The treatment with P. oleracea L. extracts downregulated MMP (MMP-1, -2, and -9) production and upregulated type I procollagen expression in UVB-induced HaCaT cells. Furthermore, treatment with the extracts decreased UVB-induced ROS generation and increased the expression of antioxidant enzymes, such as SOD-1 and OH-1, through the Nrf-2 pathway. Taken together, these results suggest that P. oleracea L. extracts could be a potential cosmeceutical agent for the prevention of skin photoaging or photodamage.

Disulfiram Suppresses Invasive Ability of Osteosarcoma Cells Via the Inhibition of MMP-2 and MMP-9 Expression

  • Cho, Hyun-Ji;Lee, Tae-Sung;Park, Jae-Bok;Park, Kwan-Kyu;Choe, Jung-Yoon;Sin, Doo-Il;Park, Yoon-Yub;Moon, Yong-Suk;Lee, Kwang-Gill;Yeo, Joo-Hong;Han, Sang-Mi;Cho, Young-Su;Choi, Myeong-Rak;Park, Nam-Gyu;Lee, Yun-Sik;Chang, Young-Chae
    • BMB Reports
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    • v.40 no.6
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    • pp.1069-1076
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    • 2007
  • Cancer cells, characterized by local invasion and distant metastasis, are very much dependant on the extracellular matrix. The expression of matrix metalloproteinases (MMPs) has been implicated in the invasion and metastasis of cancer cells. In this study, we reported the effects of disulfiram, a clinically used anti-alcoholism drug, on tumor invasion suppression, as well as its effects on the activity of MMP-2 and MMP-9 in human osteosarcoma cells (U2OS). Disulfiram has been used for alcohol aversion therapy. However, recent reports have shown that disulfiram may have potential in the treatment of human cancers. Herewith, we showed that the anti-tumor effects of disulfiram, in an invasion assay using U2OS cells and that disulfiram has a type IV collagenase inhibitory activity that inhibits expression of genes and proteins responsible for both cell and non-cell mediated invasion on pathways. In conclusion, disulfiram inhibited expression of MMP-2 and MMP-9 and it regulated the invasion of human osteosarcoma cells. These observations raise the possibility of disulfiram being used clinical for the inhibition of cancer invasion.

Effectiveness and Preparation of Microsome containing Fermented Squalene (발효 스쿠알렌을 함유한 마이크로좀의 제조 및 효능효과)

  • Kim, Ye-Jin;Kim, Tae-Hyun;Cho, Heui-Kyoung;Seong, Nak-Jun;Kim, In-Young;Yoo, Kwang-Ho;Kim, Young-Ho
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.5
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    • pp.1159-1170
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    • 2020
  • In this study, to improve the stability of fermented squalene developed using microorganisms, Microsome-SQ20 was prepared, and its physical behavior, properties, and efficacy were studied. The appearance of Microsome-SQ20 was a transparent liquid, no smell, and had a specific smell. The color was a transparent liquid, and the specific gravity was 0.928 and the pH was 5.82 (20% solution), forming a nano-emulsion suitable for use in cosmetics. It was confirmed that the content of the main component of squalene was 20.05%, which was stably sealed. The particle size measured by 0.1% aqueous solution of Microsome-SQ20 was 134.8 nm to obtain a bluish emulsified phase. The antioxidant effects of F-SQ and MF-SQ by DPPH radicals were 80.72% and 81.5%, respectively, showing superior effects compared to L-ascorbic acid. The cell viability of squalene (SQ), fermented squalene (F-SQ) and microsome squalene (MF-SQ) was at 10 ppm, respectively, showing 121.2%, 150.3%, and 129.9% cell viability. It was found that SQ, F-SQ, and MF-SQ had an elastase inhibitory ability of 8.7%, 10.33% and 8.7% at 10 ppm, respectively. In addition, the inhibitory ability of MMP-1 was 1.55%, 41.44%, 31.79% at 10 ppm for SQ, F-SQ, and MF-SQ, respectively, indicating that F-SQ significantly reduced the MMP-1 expression.

Purinergic Receptors Play Roles in Secretion of Rat von Ebner Salivary Gland

  • Kim, Sang-Hee;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • v.31 no.4
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    • pp.141-148
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    • 2006
  • The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

Antioxidation and Anti-photoaging Effects of White Taraxacum Coreanum Extract by Lactobacillus plantarum (Lactobacillus plantarum에 의한 흰 민들레 발효물의 항산화 및 광노화 억제 효과)

  • Hong, Ji Woo;Park, Ha Young;Kim, Jun Hee;Yeom, Suh Hee;Kim, Jin Woo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.4
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    • pp.554-562
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    • 2021
  • Lactobacillus plantarum SM4, a strain producing β-glucosidase, was isolated from kimchi and fermented with white Taraxacum coreanum to enhance the production of bioactive compounds. The total polyphenol content(TPC), total flavonoid content(TFC), radical scavenging activity(RSA), tyrosinase inhibitory activities(TIA), and collagenase inhibitory activities(CIA) were measured to evaluate the skin whitening and anti-wrinkle effects of the fermented product. The TPC of fermented white T. coreanum was 41.8±0.26 mg GAE/g DW, which was approximately two times higher than the hot-water extraction of 21.4±0.67 mg GAE/g DW. RSA, an indicator of antioxidant activity, was 65.6±4.7% in fermentation, which is four times higher than that of the hot-water extract. TAI and CAI, which are indicators of the whitening and anti-wrinkle effects, were 87.9±4.73% and 66.7±3.48%, respectively, which were 2.4 and 1.5 times higher than those of hot-water extraction. When comparing the UVA(320 nm) protection effects of fermented and hot-water extraction, the fermented white T. coreanum showed higher protection with an absorption rate of 64.7% and 15.2%, respectively. The white T. coreanum fermented product showed higher bioactive properties and improved skin whitening, anti-wrinkle, and UV protection effects through the production of β-glucosidase from L. plantarum SM4.

Functional Characterization of the Extracts from Nipa Palm, Molokhia, and Finger Root for Cosmetic Ingredients (니파팜, 몰로키아, 핑거루트 추출물의 화장품 소재로서의 기능적 특성 분석)

  • Jun, Yue Jin;Lee, Sohyun;Heo, Sojeong;Jin, Byung Suk
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.821-829
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    • 2019
  • This study was conducted to evaluate the applicability of the extracts from nipa palm, molokhia, and finger root in functional cosmetics as a natural active ingredient. The extracts were obtained through the processes of heating under reflux with ethanol, filtration, concentration, and freeze-drying. UV absorption and blocking effects of the extracts were examined by using the UV-vis spectrophotometer equipped with an integrating sphere. Antioxidant activity and its stability between the extracts were compared using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Also, total polyphenol content in the extracts was determined quantitatively using the Folin-Ciocalteu reagent, with gallic acid as the standard. Antibacterial activity of the extracts was investigated by the disc diffusion test against Staphylococcus aureus (gram-positive) and Escherichia coli (gram-negative). Finally, collagenase inhibitor assay was performed to examine the anti-wrinkle effect of the extracts. From the results of this study, the extract of nipa palm showed the potential for use in cosmetics as an antioxidant and anti-wrinkle agent, and the extract of finger root as a sunscreen and antibacterial agent.

THE STUDY ON EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-AA, BB ON THE CELLULAR ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Platelet-derived growth factor-AA, BB가 치주인대세포의 세포활성에 미치는 영향에 대한 연구)

  • Oh, Sang-Deok;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.303-320
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    • 1994
  • Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

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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.