• Title/Summary/Keyword: collagen assay

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ROS Scavenging Effect and Cell Viability of Opuntia humifusa Extract on Osteoblastic MC3T3-E1 Cells (천년초 추출물이 조골세포의 증식과 ROS소거능에 미치는 영향)

  • Hwang, Hyun-Jung;Jung, Bok-Mi;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1752-1760
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    • 2011
  • In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Effect of Hijikia fusiforme Fractions on Proliferation and Differentiation in Osteoblastic MC3T3-E1 Cells (톳 분획물이 조골세포의 증식 및 분화에 미치는 영향)

  • Jeon, Min-Hee;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.21 no.2
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    • pp.300-308
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    • 2011
  • Osteoporosis is a disease involving a decrease in bone mineral density and increased risk of fractures. Osteoblast and osteoclast activities are important for bone formation. The MC3T3-E1 osteoblastic cell line is a well-accepted model of osteogellsis in vitro. Hijikia fusiforme is a kind of edible brown seaweed that grows mainly in the Northwest Pacific region, including the countries of Korea, Japan and China, and it has been widely used as a medicinal and health food in Korea. In this study, by using osteoblasts, the effects of Hijikia fusiforme fractions on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and mineralization of cells were investigated. Hijikia fusiforme were subjected to fractionation by using hexane, methanol, butanol and aqueous. Proliferation of the MC3T3-E1 osteoblastic cells that were treated with Hijikia fusiforme fractions increased by approximately 120%. Regarding effects of Hijikia fusiforme fractions on ALP activity, 1 ${\mu}g$/ml butanol fraction showed the highest activity. The synthesis of collagen increased significantly in response to treatment with Hijikia fusiforme fractions, with the exception of the hexane fraction. Moreover, mineralization in the MC3T3-E1 cells that were treated with 100 ${\mu}g$/ml butanol fraction increased by 281%. Also, when 100 ${\mu}g$/ml aqueous fraction was added, mineralization increased by 240%. These results indicate that Hijikia fusiforme fractions have anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Clinical Use of PFA®-100 in Pre-surgical Screening for Platelet Function Test (수술 전 혈소판 기능 검사를 위한 PFA®-100의 임상적 이용)

  • Kim, Sung-Man;Yang, Seung-Bae;Lee, Jehoon
    • Korean Journal of Clinical Laboratory Science
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    • v.41 no.1
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    • pp.1-5
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    • 2009
  • The Platelet Function Analyzer (PFA)$^{(R)}$-100 measures the ability of platelets activated in a high-shear environment to occlude an aperture in a membrane treated with collagen and epinephrine (CEPI) or collagen and ADP (CADP). The time taken for the flow across the membrane to stop (closure time, CT) is recorded. The aim of this study was to assess the potential of the PFA$^{(R)}$-100 as a primary clinical screening tool using the wide spectrum of clinical samples assessed for platelet function as well as to perform the optimal algorithm for the use of PFA$^{(R)}$-100. We established the reference interval in 460 hospital inpatients defined as having normal platelet function based on classical laboratory tests. The reference interval by using the range $5^{th}$ and $95^{th}$ percentile was 84~251 seconds for males CEPI-CT and 85~249 seconds for females CEPI-CT. A total of 1,200 inpatients were enrolled to identify impaired hemostasis before surgical interventions. The abnormal group showing prolonged CEPI-CT was 303 cases (18.9%). Only 3 cases had both abnormal CEPI-CT and CADP-CT. Several factors including sample errors, drugs, hematologic abnoralities were contributed to unexpected prolonged CEPI-CT for screening test. The von Willebrand factor (vWF:Ag) assay was performed only in one patient to verify the algorithm for the use of PFA$^{(R)}$-100. The PFA$^{(R)}$-100 was sensitive and rapid method for primary screening test of platelet dysfunction, so we can substitute it for the bleeding time in routine clinical practice.

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Antifibrotic Effect of Curcumin in TGF-β1-Induced Myofibroblasts from Human Oral Mucosa

  • Zhang, Shan-Shan;Gong, Zhao-Jian;Li, Wen-Hui;Wang, Xiao;Ling, Tian-You
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.1
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    • pp.289-294
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    • 2012
  • Background: Myofibroblasts play an important role in the development of oral submucous fibrosis (OSF). In the current study, we investigate the effect of curcumin on growth and apoptosis of myofibroblasts derived from human oral mucosa. Methods: Myofibroblasts were generated by incubating fibroblasts, obtained from human oral mucosa, with transforming growth factor-${\beta}1$ (TGF-${\beta}1$). MTT, PI staining, and FACS assays were used to investigate curcumin's effect on proliferation and cell cycle of fibroblasts and myofibroblasts. Annexin V/PI binding and FACS assays were used to examine apoptosis of myofibroblasts, Western blotting to determine the levels of Bcl-2 and Bax, and enzyme-linked immunosorbant assay was employed to examine the levels of collagen type I and III in the supernatants of myofibroblasts. Results: Curcumin inhibits proliferation of fibroblasts and myofibroblasts; it also disturbs the cell cycle, induces apoptosis and decreases the generation of collagen type I and III in myofibroblasts, which are more sensitive to its effects than fibroblasts. Curcumin induces apoptosis in myofibroblasts by down-regulating the Bcl-2/ Bax ratio. Conclusion: Our results demonstrate the antifibrotic effect of curcumin in vitro. It may therefore be a candidate for the treatment of OSF.

Relaxing Effects of Acanthopanacis Cortex through NO Production and PDE-5 Inhibition in Corpus Cavernosum (오가피의 NO 생성과 PDE-5 억제를 통한 음경해면체 이완효과)

  • Kim, Ho Hyun;Park, Sun Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.1
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    • pp.52-58
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    • 2017
  • This study was aimed to examine relaxing effects of Acanthopanacis cortex(AC) through nitric oxide(NO) production and phosphodiesterase type 5(PDE-5) inhibition in corpus cavernosum. In order to define the relaxation effects of AC extract, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}8mm$ sized strip. AC extract ($0.01-3.0mg/m{\ell}$) were treated in contracted strips induced by phenylephrine(PE) and $N{\omega}$-nitro-L-arginine (L-NNA) was treated before AC extract-treated. And calcium chloride($Ca^{2+}$) 1 mM was infused into precontracted strips after pretreatment of AC extract in $Ca^{2+}-free$ krebs-ringer solution. When AC extract was applied to human umbilical vein endothelial cell(HUVEC), cell viability was measured by MTT assay, and NO concentration was measured by Griess reagent system. Ratio of smooth muscles to collagen fibers and eNOS, PDE-5 positive reaction were measured by histochemical and immunohistochemical process on mice corpus cavernosum. AC extract significantly affected relaxion of the cavernous strips, and the pretreatment of L-NNA inhibited AC extract-induced relaxation. Contraction induced by the addition of $Ca^{2+}$ was inhibited by treatment with the AC extract in $Ca^{2+}-free$ solution. In AC group, NO concentration, ratio of smooth muscle to collagen fibers, and eNOS positive reaction were increased, PDE-5 positive reaction was decreased compared to PE group. As a result of the above experiment, it was thought that AC extract inhibits the inflow of extracellular $Ca^{2+}$ by activating cGMP through the increase of eNOS / NO and the decrease of PDE-5 which inhibits cGMP activity, in the corpus cavernosum.

Relationship Between Depression Anxiety Stress Scale (DASS) and Urinary Hydroxyproline and Proline Concentrations in Hospital Workers

  • Lee, Keou-Won;Kim, Soo-Jeong;Park, Jae-Beom;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.44 no.1
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    • pp.9-13
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    • 2011
  • Objectives: Although increased reactive oxygen species (ROS) is caused by stress accelerates collagen degradation, there was no data on the relationship between stress and urinary hydroxyproline (Hyp) and proline (Pro), a good marker of collagen degradation. The purpose of this study was to evaluate the relationship between depression, anxiety, and stress (DAS) and concentrations of urinary Hyp and Pro. Methods: 97 hospital employees aged 20 to 58 were asked to fill out comprehensive self-administrated questionnaires containing information about their medical history, lifestyle, length of the work year, shit-work and DAS. Depression Anxiety Stress Scale (DASS) was applied to evaluate chronic mental disorders. Urine samples were analyzed using High Performance Liquid Chromatography (HPLC) with double derivatization for the assay of hydroxyproline and proline. Results: The mean value of Hyp and Pro concenturation in all subjects was $194.1{\pm}113.4\;{\mu}mol/g$ and $568.2{\pm}310.7\;{\mu}mol/g$. DASS values and urinary Pro concentrations were differentiated by sex (female > male, p < 0.05) and type of job (nurse > others, p < 0.05). In the stepwise multiple linear regressions, urinary Hyp and Pro concentrations were influenced by stress (Adjusted $r^2$ = 0.051) and anxiety and job (Adjusted $r^2$ = 0.199), respectively. Conclusions: We found that stress and anxiety were correlated with urinary Hyp and Pro concentrations. To identifying a definite correlation, further study in large populations will be needed.

도홍음(桃紅飮)이 Collagen 등(等)으로 유발(誘發)한 혈전(血栓) 및 혈액점도(血液粘度)에 미치는 영향(影響)에 관(關)한 연구(硏究)

  • Hong Ji-Seon
    • Herbal Formula Science
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    • v.6 no.1
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    • pp.199-214
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    • 1998
  • This study was performed to prove the antithrombotic effects of DoHongYeum fluid by way of experimental methods. The thrombosis was induced by injection of collagen the mixture (0.1ml/10g, 2mg/kg B.W)plus serotonin (5mg/kg B.W) into the caudal vein of rat, 2 hours after liquid extract of DoHongYeum was oral administration. The effect of the fluid of DoHongYeum was rxamined by the number of RBC and platelets, bleeding time, blood clotting time, death rate, platelet aggregation, plasmacoagulation factor activity, exvivo and fibrinolytic activity of englobulin fracture in the rats. The results were summerized as followings. 1. The number of RBC and platelets was significantly increased in DoHomeYeum group incomparison with the control group. 2. Bleeding time was significantly shortened in DHY group in comparison with the controlgroup. 3. Blood clotting time was significantly prolonged in DHY group in comparison with the control group. 4. The death rate of mouse was inhibited in DHY group in comparison with the control group. 5. The platelet aggregation was inhibited in DHY group in comparison with the control group. 6. The prothrombin time and activated partial thromboplastin time on the test of plasmacoagulation factor activity was prolonged but was not valuable in DHY group. 7. Fibrinogen lyses time of rat was reduced and lyses area was increased in DHY group incomparison with the control group. 8. Fibrinogen lyses time of rat in vitro assay was reduced in DHY group. From the above results, it was thought that the DoHongYeum could be applied effectively in the thrombosis.

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Effects of Conjugated Linoleic Acid (CLA) on Matrix Metalloproteinase (MMP) Activity and Cell Motility in Human Colon Cancer Cell Lines (Conjugated Linoleic Acid (CLA)가 인체 대장암 세포주에서 Matrix Metalloproteinase (MMP) 활성과 세포이동성에 미치는 영향)

  • 설소미;방명희;최옥숙;윤정한;김우경
    • Journal of Nutrition and Health
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    • v.36 no.3
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    • pp.280-286
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    • 2003
  • Conjugated linoleic acid (CLA) consists of several geometric isomers of linoleic acid. CLA is found in foods derived from ruminants and exhibits strong anticarcinogenic effects in a variety of animal models. Matrix metalloproteinases (MMPs) play a key role in cancer progression. Specifically, MMP-2 and -9, which hydrolyze the basal membrane type IV collagen, are involved in the initial breakdown of collagen and basement membrane components during tumor growth and invasion. However, the effects of CLA on cancer cell motility and MMP expression and activity are not currently well known. Therefore, the present study examined whether CLA reduces the activity of MMP and cell motility in SW480 and SW620 cells, the human colon cancer cell lines. Gelatin zymography and Western blot analysis revealed that phorbol 12-myristate 13-acetate (PMA) induced the activity and protein expression of Mr 92,000 MMP-9 in both cell lines. To examine whether CLA inhibits the MMP activity, cells were incubated with 100 ngfmL PMA in the presence of various concentrations of CLA. PMA-induced MMP-9 activity was decreased by 20 $\mu$ M CLA in SW480 cells, and by 10 $\mu$ M and 20 $\mu$ M CLA in SW620 cells. Results from the Hoyden chamber assay showed that cell motility was increased by PMA and that PMA-induced cell motility was significantly decreased by 20 $\mu$ M CLA in SW480 cells. These results indicate that CLA may reduce the motility and MMP activity in human colon cancer cells.

Anti-wrinkle Activity of Lindera obtusiloba Extract (생강나무 추출물의 광노화에 의한 주름형성 억제 효과)

  • Park, Keum-Ju;Park, Seung-Hee;Kim, Jae-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.4
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    • pp.317-323
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    • 2009
  • Chronic exposure of solar ultraviolet (UV) light to human skin results in photoaging with wrinkle formation. This study was performed to investigate anti-wrinkle effects of Lindera obtusiloba extract (LO) on UVB-induced wrinkle formation. We first measured cell proliferation and type I pN collagen synthesis activities in normal human dermal fibroblasts. Cell proliferation and type I pN collagen synthesis were increased by 33.8 % and 91.8 %, respectively, compared with no treatment control. SKH-1 hairless mice were topically applied 5 % LO solution for 10 weeks with UVB irradiation three times a week. After 10 weeks, a visual assessment and replica assay were performed on each mouse. According to visual assessment of close-up photos and skin replica, application of 5 % LO solution inhibited UV-induced wrinkle formation in mouse skin as compared to the vehicle-applied control mice. These results indicated that LO could protect skin wrinkle formation caused by chronic photo-irradiation in hairless mice.

Effect of Injin Fraction on Hepatic Fibrosis induced by $TGF-{\beta}1$ (인진이 $TGF-{\beta}1$ 유도성 간섬유화에 미치는 영향)

  • 신성만;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.141-155
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    • 2001
  • Objective : The aim of this study is to investigate the effect of Injin fractions on hepatic fibrosis induced by $TGF-{\beta}1$. Method : $TGF-{\beta}1$ mRNA, protein, $TGF-{\beta}1$ receptor, Smad family and PAI-I mRNA were studied in HepG2 cell, and the proliferation, connective tissue growth factor, fibronectin and collagen type I mRNA in T3891 fibroblast by quantitative RT-PCR, ELISA and thymidine incorporation assay. Results : On $TGF-{\beta}1$ mRNA and protein synthesis in HepG2, $H_2O$, butanol and hexane fractions of Injin showed inhibitory effect in a dose-dependent way. In the study on $TGF-{\beta}1$ receptor, Smad family and PAI-1 mRNA in HepG2, $H_2O$, butanol and hexane fraction of Injin showed inhibitory effect on the expression of PAI-1 in a dose-dependent way. On the proliferation of T3891 fibroblast induced by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect. In the study on the factors affected by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect on CTGF, and $H_2O$, butanol, chloroform and hexane fractions showed inhibitory effect on the expression of collagen type I, whereas no fraction showed inhibitory effect on the expression of fibronectin Conclusion : These results show that each fraction of Injin acts as a fibrosis inhibitory factor by itself or in combination, ultimately inhibiting liver cirrhosis.

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