• Title/Summary/Keyword: Collagen peptide

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Effects of Polygoni Multiflori Radix on the Elastase, and Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts

  • Kim, Myung-Gyou;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.29 no.1
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    • pp.7-12
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    • 2014
  • Objectives : Polygoni Multiflori Radix (PMR), the roots of Polygonum multiflorum Thunberg, is used to nourish the blood and yin and used for preventing premature greying of the hair. There are some articles on its preventing effects on the melanogenesis. However, there is no report about its effects on the collagen and elastin. The present study was designed to investigate its effects on collagen metabolism and elastase activity. Methods : The effects of PMR on type I procollagen production and collagenase activity in human normal fibroblasts Hs68 after UVB (312 nm) irradiation were measured by ELISA method. Cells were pretreated with the PMR for 24 hours prior to UVB irradiation. After UVB irradiation, cells were retreated with the sample and incubated for additional 24 hours. The amount of collagen type I was measured with a procollagen type I C-peptide assay kit. The activity of collagenase was measured with a MMP-1 human biotrak ELISA system. The elastase activities after treatment of PMR were measured as well. Results : In the present study, the collagen production was not increased. However, the increased collagenase activity after UVB damage was significantly recovered to $50.2{\pm}14.5%$, $8.2{\pm}3.1%$, and $10.0{\pm}3.3%$ (10, 30, and $100{\mu}g/ml$). The elastase activities (10, 100, and $1000{\mu}g/ml$) significantly reduced to $75.2{\pm}5.2%$, $40.3{\pm}1.2%$, and $27.0{\pm}1.9%$, respectively. Conclusion : PMR showed the inhibitory effects on collagenase and elastase activity. These results suggest that PMR may have potential as an anti-aging ingredient in cosmetic herbal treatment.

Anti-Wrinkle Effect of Schizandra chinensis Baillon Fermented with Lactobacillus plantarum (오미자 유산균 발효물의 주름개선 효과)

  • Lee, Jung Hee;Kim, Jong Im;Choi, Hwa Jung;Lee, Jung Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.4
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    • pp.365-371
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    • 2014
  • To identify new active anti-wrinkle ingredients, this study investigated the anti-wrinkle effects of Schizandra chinensis Baillon fermented with Lactobacillus plantarum (SCF) by assessment of cytotoxicity of human dermal fibroblast, collagen biosynthesis, matrix metalloproteinase-I (MMP-1) inhibition and elastase inhibition. S. chinensis was fermented with L. rhamnosus for 1 day at $37^{\circ}C$. The cytotoxicity of SCF was evaluated by a cytopathic effect reduction method. Effects on collagen biosynthesis and matrix metalloproteinase-I (MMP-1) of SCF were evaluated by previous reported method using procollagen type-IC peptide EIA kit and Matrix Metalloproteinase-1 Biotrack activity Assay Kit, respectively. Elastase inhibition assay was conducted by reaction of enzyme and substrate using N-Suc-$(Ala)_3$-nitroanilide as the substrate. As the results, SCF didn't show cytotoxicity against human dermal fibroblast at concentration of $100{\mu}g/mL$. Also, SCF was increased collagen synthesis and showed inhibitory effect of MMP-1 (p < 0.05). In the elastase inhibition assay, the $IC_{50}$ of SCF was $36.4{\mu}g/mL$. Therefore, our results indicated that SCF possesses anti-wrinkle effects and can be used practically for anti-wrinkle care of skin.

Application as a Functional Cosmetic Ingredient of Carrot Glycoprotein (당근 당 단백질의 기능성 화장품 소재로서의 응용성)

  • Lee, Mi-Jin;Jang, Boo-Sik;Jeong, Noh-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.2
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    • pp.257-267
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    • 2012
  • The present studies were undertaken to compared application as cosmetic ingredients of carrot glycoprotein(CG) manufactured by carrot and it's application for raw material of beauty ingredient with those of scale collagen peptide(SCP). CG and SCP apply functionality of each cream did not have fading, smell change, creaming effect and cohesion, that means the CG's properties turned out to be very stable in $5^{\circ}C$, $25^{\circ}C$. Trans epidermal water loss content was significantly lower in CG and amount of water contained in skin was significantly higher in CG. These results suggest that cream containing CG turned to be very effective in improving wrinkles excellent humid-protection as well as SCP to skin.

Histone deacetylase inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats

  • Lee, Eunjo;Song, Min-ji;Lee, Hae-Ahm;Kang, Seol-Hee;Kim, Mina;Yang, Eun Kyoung;Lee, Do Young;Ro, Seonggu;Cho, Joong Myung;Kim, Inkyeom
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.5
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    • pp.477-485
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    • 2016
  • CG200745 is a novel inhibitor of histone deacetylases (HDACs), initially developed for treatment of various hematological and solid cancers. Because it is water-soluble, it can be administered orally. We hypothesized that the HDAC inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in deoxycorticosterone acetate (DOCA)-induced hypertensive rats. For establishment of hypertension, 40 mg/kg of DOCA was subcutaneously injected four times weekly into Sprague-Dawley rats. All the rats used in this study including those in the sham group had been unilaterally nephrectomized and allowed free access to drinking water containing 1% NaCl. Systolic blood pressure was measured by the tail-cuff method. Blood chemistry including sodium, potassium, glucose, triglyceride, and cholesterol levels was analyzed. Sections of the heart were visualized after trichrome and hematoxylin and eosin stain. The expression of hypertrophic genes such as atrial natriuretic peptide A (Nppa) and atrial natriuretic peptide B (Nppb) in addition to fibrotic genes such as Collagen-1, Collagen-3, connective tissue growth factor (Ctgf), and Fibronectin were measured by quantitative real-time PCR (qRT-PCR). Injection of DOCA increased systolic blood pressure, heart weight, and cardiac fibrosis, which was attenuated by CG200745. Neither DOCA nor CG200745 affected body weight, vascular contraction and relaxation responses, and blood chemistry. Injection of DOCA increased expression of both hypertrophic and fibrotic genes, which was abrogated by CG200745. These results indicate that CG200745 attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats.

Inhibitory Effect of Artemisiae Capillaris Herba on Fibrogenesis in Rat Hepatic Stellate Cells (인진(茵蔯)이 간성상세포의 섬유화 억제에 미치는 영향)

  • Kim, Seong-A;Woo, Hong-Jung;Kim, Young-Chul;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.177-188
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    • 2008
  • Objectives : This study was performed to investigate the anti-fibrogenic effect of Artemisiae Capillaris Herba on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Artemisiae Capillaris Herba extract for 24 hours. The extraction was done either with distilled water or 50% EtOH. After the treatment, cell viability, proliferation, procollagen levels and the mRNA of the collagen type 1a2 and ASMA were measured by using MTT assay, BrdU assay, RT-PCR, and Procollagen Type I C-peptide EIA Kit. Results : The viability and proliferation of the hepatic stellate cells were decreased as the concentration increased. The mRNA expression decreased consistently with the volume of the secreted procollagen with the extraction made with distilled water, which indicates the herb has inhibitory effect on fibrogenesis of the liver by regulating one of the fibrosis associated genes in transcription. However, it increased in 50% EtOH extraction, which shows that a more stable reaction is expected of the extraction made with distilled water than the extraction made with 50% EtOH. The production of procollagen was decreased by a low-concentration treatment with Artemisiae Capillaris Herba, but increased by a high concentration. It seemed that the cells were responding to Artemisiae Capillaris Herba in low- concentrations, thus producing small amounts of collagen. When the drug was administered at high enough concentration to give direct toxicity to cells, the ability of cells to produce collagen was activated, and the overproduction of collagen was observed as an undesirable results. Conclusion : These results suggest that Artemisiae Capillaris Herba is beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis when extracted with water in the proper concentrations.

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Isolation and Characterization of Recombinant Vibrio parahaemolyticus Collagenase (재조합 Vibrio parahaemolyticus 콜라겐분해효소의 분리 및 특성 분석)

  • 차재호;김수광;전인준;이재원
    • Journal of Life Science
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    • v.13 no.3
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    • pp.308-313
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    • 2003
  • The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

Markers of Collagen Change in Chronic Secondary Renal Disease Model in Rat (만성 속발성 신질환 모델동물에서 콜라젠 변화의 지표)

  • 남정석;김기영;이영순
    • Toxicological Research
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    • v.12 no.2
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    • pp.213-221
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    • 1996
  • In order to develop a suitable secondary renal disease model and diagnostic markers of renal disease in the rat, the change of PIIIP (aminoterminal procollagen III peptide) in serum and hydroxyproline levels in the renal tissue that reflect the synthesis of extracellular matrix (ECM) during development of experimental renal diseases were observed. Two types of experimental primary diseases, diabetes mellitus administrated by streptozotocin (STZ, 75 mg/kg, i.p.) and liver cirrhosis produced by bile duct ligation/scission (BDL/s) operation, were induced. The hydroxyproline level increased according to the high PIIIP and NCl(carboxyterminal procollagen IV peptide) in Western blot analysis as early as 1 week in the STZ treated-rat kidney. Increased renal ECM was observed at 15 weeks in STZ and BDL/s model under the microscopic examination. High PAS positive reaction was found in capillary basement membrane in STZ treated-rats and mesangium in BDL/s operated rats at this time, showing the histological characteristics of diabetic nephropathy and cirrhotic glomerulonephritis in human, respectively. Such secondary renal failure were supported by additional tests including urinalysis and renal function test. The serum PIIIP detected by ELISA was a useful parameter to estimate synthesis rate of renal ECM during development of renal disease without extrarenal fibrosis i.e. liver cirrhosis in rats. This study is proposed that STZ treatment or BDL/s operation may be a suitable experimental animal model for the induction and development of chronic secondary renal diseases. Morover, it was found that hydroxyproline level in renal tissues was a good parameter of the change of renal ECM at the early stage of the diseases without apparent histological changes. Especially, serum PIIIP could be a choice as a diagnostic or prognostic marker during the development of renal diseases in rats.

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Metformin Inhibits Isoproterenol-induced Cardiac Hypertrophy in Mice

  • Cha, Hye-Na;Choi, Jung-Hyun;Kim, Yong-Woon;Kim, Jong-Yeon;Ahn, Myun-Whan;Park, So-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.377-384
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    • 2010
  • The present study examined whether metformin treatment prevents isoporterenol-induced cardiac hypertrophy in mice. Chronic subcutaneous infusion of isoproterenol (15 mg/kg/24 h) for 1 week using an osmotic minipump induced cardiac hypertrophy measured by the heart-to-body weight ratio and left ventricular posterior wall thickness. Cardiac hypertrophy was accompanied with increased interleukin-6 (IL-6), transforming growth factor (TGF)-${\beta}$, atrial natriuretic peptide (ANP), collagen I and III, and matrix metallopeptidase 2 (MMP-2). Coinfusion of metformin (150 mg/kg/24 h) with isoproterenol partially inhibited cardiac hypertrophy that was followed by reduced IL-6, TGF-${\beta}$, ANP, collagen I and III, and MMP-2. Chronic subcutaneous infusion of metformin did not increase AMP-activated protein kinase (AMPK) activity in heart, although acute intraperitoneal injection of metformin (10 mg/kg) increased AMPK activity. Isoproterenol increased nitrotyrosine levels and mRNA expression of antioxidant enzyme glutathione peroxidase and metformin treatment normalized these changes. These results suggest that metformin inhibits cardiac hypertrophy through attenuating oxidative stress.

Inhibition of Prolyl 4-Hydroxylase by Oxaproline Tetrapeptides In Vitro and Mass Analysis for the Enzymatic Reaction Products

  • Moon Hong-sik;Begley Tedhg P.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.61-64
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    • 2000
  • A series of 5-oxaproline peptide derivatives was synthesized and evaluated for its ability to inhibit the prolyl 4-hydroxylase in vitro. Structure-activity studies show that the 5-oxaproline sequences, prepared by the 1,3-dipolar cycloaddition of the C-methoxycarbonyl-N-mannosyl nitrone in the presence of the ethylene, are more active than the corresponding proline derivatives. Prolyl 4-hydroxylase belongs to a family of $Fe^{2+}-dependent$ dioxygenase, which catalyzes the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in -Gly-Xaa-Pro-Gly- of procollagen chains. In this paper we discover the more selective N-Cbz-Gly-Phe-Pro-Gly-OEt $(K_m\;=\;520\;{\mu}M)$ sequences which are showed stronger binding than others in vitro. Therefore, we set out to investigate constrained tetrapeptide that was designed to mimic the proline structure of pep tides for the development of prolyl 4-hydroxylase inhibitor. From this result, we found that the most potent inhibitor is N-Dansyl-Gly-Phe-5-oxaPro-Gly-OEt $(K_i\;=\;1.6\;{\mu}M)$. This has prompted attempts to develop drugs which inhibit collagen synthesis. Prolyl 4-hydroxylase would seem a particularly suitable target for antifibrotic therapy.

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