• Title/Summary/Keyword: collagenase activity

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The Isoflavonoid Constituents and Biological Active of Astragalus Radix by Fermentation of β-glucosidase Strains (β-glucosidase 활성 균주 발효에 의한 황기 Isoflavonoid 성분변화 및 생리활성)

  • Kim, Chul Joong;Choi, Jae Hoo;Seong, Eun Soo;Lim, Jung Dae;Choi, Seon Kang;Yu, Chang Yeon;Lee, Jae Geun
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.5
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    • pp.371-378
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    • 2020
  • Background: In this study, the radix of Astragalus membranaceus Bunge extract fermented by Saccharomyces cerevisiae, Weissella cibaria, and Pediococcus pentosaceus to increase the levels of isoflavonoid aglycone contents. Methods and Results: In order to change the in isoflavonoids, we fermented the radix of A. membranaceus extracts with microorganisms that have β-glucosidase activity. Besed on the β-glucosidase activity, we selected three strains, Weissella cibaria, Pediococcus pentosaceus, and Saccharomyces cerevisiae. HPLC analysis revealed that the levels of isoflavonoid aglycones were increased in all fermentation cases, and the extracts fermented by S. cerevisiae showed the highest levels of isoflavonoid aglycones. We evaluated the antioxidant activity, anti-wrinkle effects and whitening effects of the S. cerevisiae-fermented extracts using the DPPH assay, tyrosinase inhibition activity assay, and collagenase inhibition activity assay. We confirmed higher activity in S. cerevisiae-fermented extracts than in control, with the half maximal inhibitory concentration (IC50) value of 565.1 ± 59.1 ㎍/㎖ in DPPH radical scavenging activity, tyrosinase inhibition rate of 78.4 ± 0.9%, and collagenase inhibition rate of 83.8 ± 1.1%. Conclusions: We selected three stains of microorganisms showing high β-glucosidase activity, W. cibaria, P. pentosaceus and S. cerevisiae. Isoflavonoid glycones in the radix of A. membranaceus were converted to isoflavonoid aglycones by fermentation. In addition, the fermented radix of A. membranaceus exhibited antioxidant activity, anti-wrinkle effect, whitening effect and radical scavenging activity.

The Analgesic Effect and the Mechanism of Electroacupuncture on Thermal Hyperalgesia in the Rat Model of Collagenase-induced Arthritis: Mediation by Adrenergic Receptors (Collagenase-induced Arthritis Rat Model에서 Thermal Hyperalgesia에 대한 전침(電鍼)의 진통효과(鎭痛效果) 및 기전연구: Adrenergic Mechanism에 대(對)한 연구(硏究))

  • Seo, Byung-Kwan;Park, Dong-Suk;Baek, Yong-Hyeon
    • Journal of Acupuncture Research
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    • v.28 no.2
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    • pp.57-67
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    • 2011
  • 목적 : Collagenase-induced osteoarthritis(OA) 동물 모델에서 전침의 adrenergic mechanism을 연구하고자 한다. 방법 : Collagenase-induced arthritis(CIA)를 유발하기 위하여 5주령의 male Sprague-Dawley rat의 뒷다리 좌측 무릎 관절에 0.05ml의 4mg/ml collagenase solution을 intra-articular 주입하고, 다시 4일 후에 같은 부위에 같은 농도의 collagenase solution을 intra-articular boosting injection 시행한 뒤, gross, histopathological features 및 biomarker activity 변화를 관찰하였다. 예비실험을 통하여 CIA rat model에서 진통효과를 발휘하는 것으로 확인한, 족삼리(足三里) ($ST_{36}$)에 대한 저빈도 train pulse EA stimulation (2Hz, 0.07 mA, 0.3ms)을 침치료 방법으로 적용하였다. 전침의 진통기전을 확인하기 위하여, ${\alpha}1$-adrenergic antagonist (prazosin, 1 mg/kg, i.p.), ${\alpha}2$-adrenergic receptor antagonist (yohimbine, 2mg/kg, i.p.), ${\alpha}1$-adrenergic receptor agonist(phenylephrine, 2mg/kg, i.p.), ${\alpha}2$-adrenergic receptor agonist(clonidine, $40{\mu}g$/kg, i.p.)을 전침시행 20분 전에 복강 내로 전처치하였다. Tail flick unit(Ugo Basile Model 7360)을 이용하여 열자극에 대한 통증역치를 측정하였다. 결과 : 퇴행성관절염 징후(gross, histopathological features)와 통증역치의 변화가 최대값을 나타내는 CIA 유발 4주차에 저빈도 전침자극(train pulse, 2Hz, 0.07mA, 0.3ms)을 족삼리($ST_{36}$)에 적용하였으며, 족삼리 전침의 진통효과는 ${\alpha}2$-adrenergic receptor antagonist(yohimbine, 2mg/kg, i.p.)전처치에 의해 억제되었으나, ${\alpha}1$-adrenergic antagonist(prazosin, 1 mg/kg, i.p.)전처치에는 억제되지 않았다. 또 ${\alpha}2$-adrenergic receptor agonist(clonidine, $40{\mu}g$/kg, i.p.)의 전처치를 통하여 유의한 synergistic analgesic effect가 관찰되었으나, ${\alpha}1$-adrenergic receptor agonist(phenylephrine, 2mg/kg, i.p.)의 전처치는 전침의 진통효과에 synergistic effect를 미치지 않는 것으로 나타났다. 결론 : 저빈도 족삼리 전침은 CIA로 유발된 염증성 통증에 대하여 진통효과를 발휘하며, 이는 ${\alpha}2$-adrenergic receptor에 의하여 매개되는 것으로 보이며 ${\alpha}1$-adrenergic receptor는 영향을 미치지 않는 것으로 사료된다.

EFFECT OF VARIOUS CYTOKINES ON THE PRODUCTION OF PROSTAGLANDIN $E_2$, LEUKOTRIENE $B_4$ AND COLLAGENASE IN HUMAN PERIODONTAL LIGAMENT FIBROBLASTS IN VITRO (수종의 cytokine이 사람 치주인대 섬유아세포의 prostaglandine $E_2$, leukotriene $B_4$ 및 collagenase 생산에 미치는 영향)

  • Kim, Jung-Ho;Suhr, Cheong-Hoon
    • The korean journal of orthodontics
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    • v.24 no.4 s.47
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    • pp.871-883
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    • 1994
  • This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

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Purification and Characterization of a Collagenase from the Mackerel, Scomber japonicus

  • Park, Pyo-Jam;Lee, Sang-Hoon;Byun, Hee-Guk;Kim, Soo-Hyun;Kim, Se-Kwon
    • BMB Reports
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    • v.35 no.6
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    • pp.576-582
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    • 2002
  • Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.

Analysis on Antioxidant Activity and Agronomic Characteristics of Extract from Smilacis Chinae Radix

  • Hyeon Mi Jo;Sin Park;Eun Bi Choi;In-Ho Choi
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.313-313
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    • 2022
  • The Smilacis chinae Radix refers to the root of Smilax chinae L distributed in mountain and filed of Korea, and it is a vine shrub in the Lilaceae family, called Berchemia berchemiaefolia, and is referred to as Smilacis chinae Radix in it's a natural medicine name. Antibacterial, inflammatory, and antioxidant activity were studied in Smilacis chinae Radix. In this study, biological activities such as antioxidant (DPPH, ABTs, TPC), cytotoxicity, wrinkle improvement, and whitening improvement to increase the utilization value of Smilacis chinae Radix and identify the botanical value. Therefore, we tried to explore the applicability of Smilacis chinae Radix as a functional cosmetic material. Smilacis chinae Radix (SCR) was dried and extracted with ethanol. In order to measure the biological activity of the SCR, antioxidant activity, inhibition activities of collagenase, tyrosinase and cell viability were measured. The DPPH (1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity in the extract with a concentration of 400㎍/mL is 91.22% ± 0.41%%. ABTs (2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity in the extract with a concentration of 400㎍/mL is 99.60% ± 0.03%. Total polyphenol contents (TPC) are 0.203 ± 0.05 mg GAE/mg Ext when SCR was lmg/mL. And the Cell viability for HaCaT derived human keratinocyte and Raw264.7, a mouse-derived macrophage was determined using the MTT assay. When cell was treated with 100㎍/mL of SCR, HaCaT cell showed cell viability of 78.09 ± 0.1% and Raw264.7 cell showed cell viability of 91.88 ± 0.42%. From the above results, we have shown the possibility that the CSR have antioxidant ability, inhibition activity of collagenase and tyrosinase and cell safety ability which can be useful in a functional cosmetic material.

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The Coffee Sliver Skin Extracts from Coffee Beans Exhibited Cosmetic Properties with Antioxiant Activity and Inhibitory Effects for Elastase, Collagenase and Tyrosinase (커피 은피 추출물의 항산화 효과와 엘라스타제, 콜라게나제 및 티로시나제 저해효과)

  • Lee, Kyung Eun;Son, Sang Hyeok;Kang, Sang Gu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.39-48
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    • 2018
  • The coffee silver skin is a part of coffee beans. We report that the coffee sliver skin extracts exhibited cosmetic properties of antioxidant, anti-winkle and whitening effects. The ethanol extracts of silver skin showed free radical scavenging activity up to 92.26% in $50{\mu}g/mL$, especially against DPPH radical and ABTS radical cation. The silver skin extracts showed inhibitory effects for tyrosinase activity and DOPA oxidation in a dose-dependent manner, suggesting the extracts retain for the whitening property in cosmetics. The coffee silver skin extracts effectively inhibited the elastase and collagenase. Cytotoxicity of the coffee silver skin extracts was measured by the colorimetric MTS assay. The viability of the human keratinocytes (HaCaT) treated with the coffee silver skin extracts was same as that of untreated cells, indicating the extracts are safe to human cells. Here, we suggest that the silver skin extracts of coffee bean could be a potential natural substance for anti-winkle, whitening, antioxidant properties for cosmetics.

Anti-Wrinkle Effects of Enzymatic Oyster Hydrolysate and Its Fractions on Human Fibroblasts (굴 효소 가수분해물과 분자량 분획물의 섬유아세포에 대한 주름 개선 효과)

  • Kim, Hyeon-Ah;Park, Si-Hyang;Lee, Su-Seon;Choi, Yeung Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1645-1652
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    • 2015
  • We investigated the anti-wrinkle efficacy of hydrolysate from oyster protein by Protamex and Neutrase for the purpose of finding materials to assist skin health originating from marine organisms. There were about 7.9% free amino acids in the oyster hydrolysate, and contents of urea, taurine, alanine, and glycine were high. Oyster hydrolysate also showed collagenase inhibitory activity and was not toxic to CCD986sk human fibroblast cells. Yield of the fractions according to the molecular weight of oyster hydrolysate was 40% for less than 1,000 Da and 60.4% for less than 5,000 Da, respectively. Antioxidative effect, procollagen production, and matrix metalloproteinase-1 inhibitory activity were highest in 1,000~3,000 Da fractions. We observed that oyster hydrolysate and its less than 5,000 Da fraction are potential functional compounds for skin health and for improving wrinkles.

α-Asarone Modulates Activity of Matrix Metalloproteinase as well as Antioxidant Activity (α-Asarone이 항산화 활성 및 기질금속단백질 분해효소 활성 조절에 미치는 영향)

  • Park, Hye-Jung;Kim, Moon-Moo
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1000-1006
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    • 2015
  • α-Asarone is the main component of Acorus gramineus, which is a widely used oriental traditional medicine. A. gramineus is known to have a variety of medicinal effects, such as anti-gastric ulcer, antiallergy and antioxidant activity. It is also known to inhibit the release of histamine. However, the mechanism of its action remains unclear in humans. In this study, the effects of α-asarone on matrix metalloproteinase (MMP) and its antioxidant effect in a cell-free system were examined in HT1080 cells. In an MTT assay, the effect of α-asarone on cell viability showed no cytotoxicity below 16 μM. In an antioxidant assay, α-asarone increased reducing power in a dose-dependent manner but not the scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. In addition, α-asarone exhibited the protective effect against DNA oxidation induced by hydroxyl radicals produced by the Fenton reaction. Furthermore, in a gelatin disk assay, α-asarone enhanced collagenase activity. It also increased the activities of MMP-2 and MMP-9 stimulated by phorbol 12-myristate 13-acetate (PMA) in a gelatin zymography. On the other hand, the activity of MMP-9 stimulated by phenazine methosulfate (PMS) but not that of MMP-2 was increased in the presence of α-asarone. These findings suggest that α-asarone could be a candidate for the prevention and treatment of pathological diseases related to oxidative stress and MMPs.

Effect of Ssanghwa-tang Extract on Antioxidant and Anti-aging Enzyme Activities (쌍화탕 추출물이 항산화효소 및 항노화관련 효소 활성에 미치는 영향)

  • Park, Ji-Young;Hwang, Jae-Gyu;Yun, Jong-Kuk;Han, Kil-Hwan;Do, Eun-Ju;Kim, Sung-Ok;Kim, Mi-Ryeo
    • The Korea Journal of Herbology
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    • v.27 no.3
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    • pp.67-74
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    • 2012
  • Objectives : The present study was designed to investigate effects of Ssanghwa-tang (Shu$\bar{a}$nghu$\bar{a}$-g$\bar{e}$ng) on oxidation/reduction reaction-related and aging-related enzymes $in$ $vitro$. Methods : We performed MTT assay, collagenase inhibition assay, elastase inhibition assay, tyrosinase inhibition assay, DPPH free radical scavenging assay, SOD-like activity and xanthine oxidase (XO) inhibition assay. Results : The 50% ethanol (EtOH) extract of Ssanghwa-tang (SHT) showed 55% inhibition of collagenase activity, and 42% inhibition of elastase activity at 1 mg/ml concentration. Also it's treatment showed 18% inhibition of tyrosinase activity, to relate whitening effect, at the same dose of 50% ethanol extract of SHT. Antioxidant activities were determined by DPPH radical scavenging, XO inhibiting activity and SOD-like activity. These scavenging, XO-inhibiting and SOD-like activities were measured in 80%, 75%, and 28% inhibitions, respectively, at a 1 mg/ml treated dose, compared to those of control. The inhibitory effects of 50% EtOH extract on aging and oxidation-related enzyme activities were higher than those of water extract and 95% EtOH extract. Conclusions : Taken together, our findings suggest that the SHT has potential and applicable benefits for development of cosmetics to have anti-aging (anti-wrinkle and whitening) and anti-oxidation functions.

Antioxidant and Antiaging Assays of Hibiscus sabdariffa Extract and Its Compounds

  • Widowati, Wahyu;Rani, Andani Puspita;Hamzah, R. Amir;Arumwardana, Seila;Afifah, Ervi;Kusuma, Hanna Sari W.;Rihibiha, Dwi Davidson;Nufus, Hayatun;Amalia, Annisa
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.192-200
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    • 2017
  • Skin aging is a complex biological process due to intrinsic and extrinsic factors. Free radical oxidative is one of extrinsic factors that induce activation of collagenase, elastase and hyaluronidase. Natural product from plants has been used as antioxidant and antiaging. This study aimed to evaluate antioxidant and antiaging properties of Hibiscus sabdariffa extract (HSE) and its compounds including myricetin, ascorbic acid, and ${\beta}$ carotene. The phytochemical of H. sabdariffa was determined using modified Farnsworth method and presence of phenols, flavonoids and tannins were in moderate content, whereas triterpenoids and alkaloids were in low content. Total phenolic content performed using Folin-Ciocalteu method, was $23.85{\mu}gGAE/mg$. Quantitative analysis of myricetin, ${\beta}-carotene$, and ascorbic acid of HSE was performed with Ultra-High Performance Liquid Chromatography (UHPLC) that shows $78.23{\mu}g/mg$ myricetin, $0.034{\mu}g/mg$ ${\beta}-carotene$, whilst ascorbic acid was not detected. HSE has lower activity on DPPH ($IC_{50}=195.73{\mu}g/mL$) compared to ${\beta}-carotene$, the lowest in ABTS assay ($IC_{50}=74.58{\mu}g/mL$) and low activity in FRAP assay ($46.24{\mu}MFe(II)/{\mu}g\;$) compared to myricetin, ${\beta}-carotene$. Antiaging was measured through inhibitory activity of collagenase, elastase, and hyaluronidase. HSE had weakest collagenase inhibitory activity ($IC_{50}=750.33{\mu}g/mL$), elastase inhibitory activity ($103.83{\mu}g/mL$), hyaluronidase inhibitory activity ($IC_{50}=619.43{\mu}g/mL$) compared to myricetin, ${\beta}-carotene$, and ascorbic acid. HSE contain higher myricetin compared to ${\beta}-carotene$. HSE has moderate antioxidants and lowest antiaging activities. Myricetin is the most active both antioxidant and antiaging activities.