• Title/Summary/Keyword: collagen assay

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Anti-osteoarthritis effects of Pomegranate, Eucommiae cortex and Achyranthis radix extracts on the primary cultured rat articular chondrocytes

  • Choi, Beom-Rak;Ku, Sae-Kwang;Kang, Su-Jin;Park, Hye-Rim;Sung, Mi-Sun;Lee, Young-Joon;Park, Ki-Moon
    • Journal of Society of Preventive Korean Medicine
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    • v.21 no.3
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    • pp.87-98
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    • 2017
  • Objectives : The objective of present study is to evaluate anti-arthritic effects of dried pomegranate concentrate powders (PCP), Eucommiae Cortex aqueous (EC) and ethanolic (ECe) extracts, Achyranthis Radix aqueous (AR) and ethanolic (ARe) extracts on the primary cultured rat articular chondrocytes. Methods : MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay was performed cytotoxic effect of test substances. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2\;(PGE_2)$ production and 5-lipoxygenase (LPO) activities, and inhibitory effects on metalloproteinase (MMP)-2 and MMP-9 activities were observed on the recombinant human interleukin $(rhIL)-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions - collagen type II, SOX9 and aggrecan. Results : As results, ECe and ARe showed obvious cytotoxicity against primary cultured rat articular chondrocytes at a dose level of 10 mg/ml, respectively. However, no obvious cytotoxic effects of PCP, EC and AR were demonstrated at a dose level of 10 mg/ml, on the primary cultured rat articular chondrocytes. In addition, treatment of LPS $50{\mu}g/ml$ induced significant increases of $PGE_2$ contents and 5-LPO activities indicating inflammatory responses of the primary cultured rat articular chondrocytes, and also decreases of cell viabilities, increases of MMP-2 and MMP-9 activities with decreases of extracellular matrix (ECM) related collagen type II, SOX9 and aggrecan mRNA expressions were observed by treatment of $rhIL-1{\alpha}$ 50 ng/ml, suggesting damages on the primary cultured rat articular chondrocytes and related ECM degradations. However, these inflammatory responses and related ECM degradations were inhibited by pretreatment of all test substances, in order of PCP > ECe > ARe > EC > AR, and $rhIL-1{\alpha}$ induced chondrocytes deaths are inhibited by treatment in order of PCP > EC > AR > ECe > ARe. Conclusions : Taken together, it is expected that mixed formulation of PCP as main components with appropriate proportion of EC and AR as additional components will be achieved a potent alternative medicinal food for osteoarthritis.

An Experimental Study of Effect on ECV 304 Cells, Platelet Rich Plasma and Rats treated with L-NAME by Boonsimgieum extract (분심기음(分心氣飮)이 고혈압 백서와 인간유래 혈관내피세포주(ECV 304)에 미치는 영향에 대한 연구)

  • Jeon, Yeon-Yi;Park, Chang-Gook;Lee, So-Yeon;Yoon, Hyeon-Deok;Shin, Wo-Cheol;Park, Chi-Sang
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.182-198
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    • 2005
  • Object : This study was designed to research whether the protection and inhibitory effects of cardiovascular diseases in L-NAME induced rat or ECV 304 cell lines through the Cell morphological pattern, Tunel assay, LDH activity, heart rate, blood pressure and immunohistochemistric analysis by Boonsimgieum water extract Methods : Nitric oxide(NO) play an important role in normal and pathophysiological cells including as a messenger molecule, neurotransmitter, microbiocidal agent, or dilator of blood vessels and artheriosclerosis, hypertension, myocardial infarction, respectively. Endothelial cell products can modulate the magnitude of a response to a vasoconstrictor, as evinced by the greater constriction after endothelium removal or NO synthesis blockade. To investigate that Boonsimgieum in the potential contribution of the levels of nitric oxide generated by endothelial nitric oxide synthase (eNOS) and the mechanisms of protection against NG-nitro-L-arginine methyl ester (L-NAME), human ECV 304 cells, which normally do not express eNOS, were expressed by L-NAME. L-NAME stimulated rat or cells were found to be resistant to injury and delayed death following the Boonsimgieum. Inhibition of nitric oxide synthesis abolished the protective effect against L-NAME, thrombin and collagen exposure. Interestingly, such effects have been observed during stimulation with agents such as phenylephrine and KCl on L-NAME mediate rats, were damaged by the NOS inhibitor L-NAME. Result : As the result of this study, In group, the anti-apoptosis and necrosis in the cardiovascular system have a potential capacity for prevented, protected and treating the diseases of cardiovascular system, against the necrosis of rat and ECV 304 cells with Caspase 3 and calpain expression by L-NAME is promoted. Conclusion : these results demonstrate neuroprotective and memory enhancing effects of ZIBU, suggesting its beneficial actions for the treatment of AD.

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Inhibitory Effects of Type IV Collagenase by Disulfiram (Disulfiram에 의한 type IV collagenase 저해효과)

  • Sin, Doo-Il;Park, Jae-Bok;Park, Kwan-Kyu;Cho, Chang-Ho;Oh, Hoon-Kyu;Choi, Chang-Hyuk;Cho, Hyun-Ji;Chang, Young-Chae
    • Journal of Life Science
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    • v.16 no.6
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    • pp.964-971
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    • 2006
  • The cancer cells, characterized by local invasion and distant metastasis, are very dependant on extracellular matrix. The expression of matrix metalloproteinases (MMPs) has been implicated in the invasion and metastasis of cancer cells. Among the human MMPs, matirx metalloproteinase-2 (MMP-2) and matrix metalloproteinse-9 (MMP-9) are key enzymes that degrade type IV collagen of the matrix. Here, we studied the effect of disulfiram, an anti-tumor compound, on the suppression of the tumor invasion and the activity of MMP-2, MMP-9 in human osteosarcoma cells (U2OS). Disulfiram had the type IV collagenase inhibitory activity, the effect of inhibition of gene and protein expression, and these inhibitions were responsible for blocking invasion through cell mediated and non-cell mediated pathways. In conclusion, disulfiram inhibited expression of MMP-2 and MMP-9, and regulated the invasion of U2OS, Caki-1 and Caski. These observations raise the possibility of clinical therapeutic applications for disulfiram used as a potential inhibitor of cancer invasion.

Preparation and Characterization of Silk Fibroin/Gelatin Hybrid Scaffolds (실크 피브로인/젤라틴 하이브리드 지지체의 제조 및 특성분석)

  • Kim, Hye-Lin;Hong, Min-Sung;Kim, Su-Jin;Jo, Han-Su;Yoo, Il-Sou;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.5
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    • pp.378-384
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    • 2011
  • Silk fibroin is a biocompatible and slowly biodegradable natural polymer. This natural polymer has excellent mechanical properties, non-toxicity, and non-immunogenic properties and has been demonstrated to support tissue regeneration. Also, gelatin is a natural material derived from collagen by hydrolysis and has an almost identical composition as that of collagen. Silk fibroin/gelatin scaffolds have been fabricated by using the freeze-drying method. To establish the scaffold manufacturing condition for silk fibroin and gelatin, we made scaffolds with various compositions of gelatin, glutaldehyde and silk fibroin. The silk fibroin/gelatin scaffolds were characterized using SEM, DSC, and water absorption ability tests. The cellular proliferation was evaluated by WST assay. These results suggested that a scaffold containing 8% of gelatin, 1% of glutaldehyde and 0.3 g of silk fibroin provided suitable characterstics for cell adhesion and proliferation. In conclusion, the silk fibroin/gelatin scaffold may serve as a potential cell delivery vehicle and a structural basis for tissue engineering.

Effects of Watercress Containing Rutin and Rutin Alone on the Proliferation and Osteogenic Differentiation of Human Osteoblast-like MG-63 Cells

  • Hyun, Hanbit;Park, Heajin;Jeong, Jaehoon;Kim, Jihye;Kim, Haesung;Oh, Hyun Il;Hwang, Hye Seong;Kim, Ha Hyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.4
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    • pp.347-352
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    • 2014
  • Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to $125.0{\pm}4.0%$ ($mean{\pm}SD$), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to $222.3{\pm}33.8%$. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to $149.2{\pm}2.8%$, and increased the degree of mineralization, a marker of the late process of differentiation, to $122.9{\pm}3.9%$. Rutin alone also increased the activity of ALP (to $154.4{\pm}12.2%$), the expression of collagen type I (to $126.6{\pm}6.2%$), and the degree of mineralization (to $112.3{\pm}5.0%$). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.

Effect of Extracellular Matrix on the Growth Behavior of Corneal Endothelial Cells to Poly(lactic-co-glycolic acid) Film (각막 내피세포 성장 거동에 대한 락타이드 글리콜라이드 공중합체 필름과 세포외 기질의 효과)

  • Kim, Eun Young;Kim, Hye Min;Song, Jeong Eun;Lee, Hyun Soo;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.702-707
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    • 2014
  • Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

A Study on Anti-oxidant and Anti-wrinkle Effect of Supercritical Fluid Extraction of Black Carrot as a Functional Cosmetic Material (기능성화장품소재로서 자색당근 초임계추출물의 항산화 및 항주름 효능 연구)

  • Kim, Ji-Su;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.11 no.12
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    • pp.236-243
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    • 2021
  • The aim of this study was to evaluate the anti-oxidant and anti-wrinkle effect of the black carrot(BC) extracted by supercritical dioxide(SC-CO2). DPPH/ABTS radical scavenging and total polyphenol contents were measured to investigate the anti-oxidant activity of the BC supercritical extract. Collagen production and MMP-1 expression were assessed in normal human dermal fibroblasts(NHDF) for anti-wrinkle activity, The black carrot extract showed the highest total phenolic content(9.037±1.123 mg GAE/g extract) and the best antioxidant properties as shown by DPPH and ABTS assay. The supercritical fluid extracts of black carrot exhibited low toxicity to NHDF cells. In addition, the supercritical fluid extracts showed MMP-1 inhibition and type I pro-collagen synthesis inducing activities in a dose-dependent manner, respectively. Therefore, these results suggest that the black carrot is a potential source of high anti-oxidative, solvent-free and anti-aging material with promising applications in cosmetic, food, and beauty-food industries.

Antiaging Activity of Mixed Extracts from Korean Medicinal Herbs on HS68 Skin Fibroblast (한약재복합 추출물의 인간피부섬유아세포 HS68에 대한 항노화 효과)

  • Shin, Dong-Chul;Kim, Gwui-Cheol;Song, Si-Young;Kim, Hee-Jin;Yang, Jae-Chan;Lee, Yong-Hwa;Kim, Bo-Ae
    • The Korea Journal of Herbology
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    • v.29 no.2
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    • pp.39-45
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    • 2014
  • Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

Anti-aging Effect of Akebia quinata Decaisne Ethanol Extract (으름덩굴 에탄올 추출물의 항노화 효과)

  • Yu Jin Kim;Soon Hyun Kwon;Ji Hyun Song;So Mi Lee;Yong Min Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.67-75
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    • 2024
  • Skin aging progresses due to external factors such as ultraviolet rays and infections. These factors cause skin fibroblasts to secrete proteolytic enzymes, matrix metalloproteinases (MMPs). MMPs induce the degradation of collagen located in the extracellular matrix, directly influencing aging. The stems of Akebia quinata Decaisne have been reported to have antioxidant and anti-inflammatory effects. However, the anti-aging effect of Akebia quinata Decaisne stem ethanol extract (AQSEE) is not known. Therefore, we studied the TNF-α-induced MMP-1 inhibitory effect in human fibroblasts. When the cell viability of AQSEE was confirmed through MTT asaay, it showed no toxicity up to 400 ㎍/mL. The inhibition of MMP-1 mRNA and protein secretion was confirmed through RT-qPCR and ELISA, and results showed a significant decrease at concentrations of 100, 200, 400 ㎍/mL. We also confirmed by Western blotting that phosphorylation of MAPKs signaling pathway and transcription factors was reduced. As a result, phosphorylation of p38, c-Jun, p65 was significantly decreased at all concentrations. DPPH and ABTS assays were performed to confirm the radical scavenging ability of AQSEE, and the results showed a significant decrease at all concentrations. The results of this study confirmed the MMP-1 inhibitory effect and radical scavenging ability, which suggests that it can be used as an anti-aging substance.

Protective Effect of Green Tea Extract, Catechin on UVB-Induced Skin Damage (녹차추출물 성분 catechin이 자외선에 의해 손상된 피부에 미치는 영향)

  • 이은희;이종권;홍진태;정경미;김용규;이선희;정수연;이용욱
    • Journal of Food Hygiene and Safety
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    • v.16 no.2
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    • pp.117-124
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    • 2001
  • The main constituent of green tea, catechins have been reported to have numberous biological anti-vites including antimutagenic, antibacterial, hypocholesterolemic, antioxidant and antitumor properties. In the present study, we examined the protective effect of catechin on UVB-induced skin damage. Catechin (3 mg/mouse) was topically treated to dorsal area of SHK-1 hairless mouse daily for 2 weeks. UVB (100 mJ/$\textrm{cm}^2$) was also treated soon after application of catechin alone or with catechin for 2 weeks. Catechin reduced UVB-induced infiltration of inflammatory cells, fibrosis of cells and collagen-fiber formation. In addition, catechin also prevented UVB-induced DNA fragmentation and apoptosis cell number, but not changed p53 level. Furthermore catechin inhibited UVB-induced cell proliferation. There results showed that catechin have preventive effect aganinst UVB-induced skin damages. and these effects could contribute to the antitumor promoters activity.

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