• 제목/요약/키워드: collagen assay

검색결과 310건 처리시간 0.031초

The study on collagen sysnthesis and collagenase inhibition assay in natural plants

  • W. J. Yang;S. J. Yang;Kim, W. H.;T. B. Kang;Park, S. S.
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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    • pp.239-240
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    • 2003
  • Type I (collagen) and procollagen are reduced in aged human skin. This reduction could result from increased degration by metalloproteinases and from reduced procollagen synthesis and skin collagenase is required for initiation of the degration of type I collagen. In the present study, we study on assay the collagen and collagenase in natural plants using the fibroblast human skin cell. We select the 15 kind of plants used to herbal and 4 kind of fraction(by methylene chloride, ethyl acetate, n-butanol, water). Among these extract, the ethyl acetate fraction from benincasa hispida/prunus persica, trichosanthes kiriowii, trogopterus xanthipes and methylene chloride fractions from benincasa hispida/prunus persica, torilis japonica and n-butanol fraction from cnidium officinale, chrysanthemum sibiricum were selected for further experiments as they exhibited distinctive amount of collagen compared to other natural extracts. These extracts were again subjected to collagenase assay test. Benincasa hispida/prunus persica extract was shown to have exellent collagen synthesis activity from result of the collagen assay test and the other extract was shown to have over 130% of collagen synthesis activity. But, in the study of collagenase assay test just only trogopterus xanthipes extract was shown to have collagenase inhibition.

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수종(數種) 한약재의 콜라겐 생합성 효과에 대한 실험적 연구 (Experimental study about the effect of several herbs on collagen synthesis)

  • 윤지은;김남권;황충연;조은희;박민철
    • 한방안이비인후피부과학회지
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    • 제23권3호
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    • pp.33-41
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    • 2010
  • Background & Objective : This study was performed to find a new anti-aging, anti-wrinkle agent. Ten kinds of hers were evaluated by proliferation, cytotoxicity, collagen synthesis tests in cultured human dermal fibroblast. Methods : MTT assay was carried out to check cell proliferation, cytotoxic effect of each herbal medicine. Collagen synthesis was investigated by the amount of propeptide using procollagen type-Ⅰ C peptide(PIP) ELISA kit(Takara). Results and Conclusions : There are no cytotoxicity in all group herbs by MTT assay. Collagen synthetic ability was greatest in LYCII FRUCTUS(LF) and was excellent in order of Rehmanniae Radix Preparat(RRP) > Nelumbinis Semen(NS) > Asparai Radix(AR) > Schizandrae Frucus(SF).

자외선 조사에 의해 노화된 섬유아세포에서 Cycloheterophyllin의 항노화 효능 (Anti-aging Effect of Cycloheterophyllin in UVA-irradiated Dermal Fibroblasts)

  • 심중현
    • 생약학회지
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    • 제50권4호
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    • pp.285-290
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    • 2019
  • This study was carried out to identify the skin anti-aging effect of cycloheterophyllin on dermal fibroblasts. To elucidate anti-aging effects of cycloheterophyllin on dermal fibroblasts, I measured cell viability, mRNA expressions, and Collagen, type I/matrix metallopeptidase 1(MMP1)-ELISA assay. In this study, I investigated the effects of cycloheterophyllin on Collagen, type I, alpha 1(COL1A1)/Collagen, type III, alpha 1(COL3A1)/MMP1/Superoxide dismutases/Catalase(CAT) mRNA expressions and Collagen, type I/MMP1 protein production. Quantitative Real-time RT-PCR showed that cycloheterophyllin increased mRNA level of COL1A1/COL3A1/CAT genes and collagen, type I protein by ELISA assay compared to UVA-treated dermal fibroblasts. Furthermore MMP1 mRNA and protein expressions were decreased by cycloheterophyllin treatment. These observations revealed that cycloheterophyllin increased anti-aging effects in dermal fibroblasts. Therefore, I identified the anti-aging effects of cycloheterophyllin, and these results showed that the cycloheterophyllin can be a considerable potent ingredient for skin anti-aging. Based on this, I anticipated further researches about cycloheterophyllin for mechanism to develop not only cosmetics but for healthcare food or medicine.

단치소요산가미방이 Dexamethasone 처리한 랫드의 두개골 세포에 미치는 영향 (Effects of DSG on Osteoblastic Cell from Rat Calvariae in the Presence of Dexamethasone)

  • 박종형;황귀서
    • 대한예방한의학회지
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    • 제10권2호
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    • pp.19-30
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    • 2006
  • It is well known that glucocorticoid may induce osteoporosis as its side effect in long-term therapy. The inhibition of osteoblast by glucocorticoid is also recognized as its action mechanism of decreased bone formation. In this study, the effect of DSG, Danchisoyosangamibang, on the differentiation and function of osteoblastic cells was investigated. The osteoblastic cells were isolated from rat calvariae using collagenase treatment. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, intracellular alkaline phosphatase (ALP) activity, bone martrix production, and cell apoptosis. DSG enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracelluar collagen synthesis were increased time dependently when the cells were treated with DSG in the presence of dexamethasone. And, DSG restored calvarial cell function decreased by dexamethasone.

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자혈양근탕(滋血養筋湯)이 부신피질호르몬에 의해 억제된 조골세포 기능에 미치는 영향 (Effects of JY on Osteoblastic Cell from Rat Calvariae in the Presence of Glucocorticoid)

  • 최정신;황귀서
    • 대한예방한의학회지
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    • 제12권2호
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    • pp.197-206
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    • 2008
  • The inhibition of osteoblast by glucocorticoid is recognized as its action mechanism of decreased bone formation. In this study, the effect of JY, Jahyulyangkeuntang, on the differentiation and mineralization of osteoblastic cells was investigated in the presence of dexamethasone. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, alkaline phosphatase(ALP) activity, bone martrix production, and cell apoptosis. JY enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracellular collagen synthesis were increased when the cells were treated with JY. And JY restored calvarial cell function decreased by dexamethasone.

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The proper concentrations of dextrose and lidocaine in regenerative injection therapy: in vitro study

  • Woo, Min Seok;Park, Jiyoung;Ok, Seong-Ho;Park, Miyeong;Sohn, Ju-Tae;Cho, Man Seok;Shin, Il-Woo;Kim, Yeon A
    • The Korean Journal of Pain
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    • 제34권1호
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    • pp.19-26
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    • 2021
  • Background: Prolotherapy is a proliferation therapy as an alternative medicine. A combination of dextrose solution and lidocaine is usually used in prolotherapy. The concentrations of dextrose and lidocaine used in the clinical field are very high (dextrose 10%-25%, lidocaine 0.075%-1%). Several studies show about 1% dextrose and more than 0.2% lidocaine induced cell death in various cell types. We investigated the effects of low concentrations of dextrose and lidocaine in fibroblasts and suggest the optimal range of concentrations of dextrose and lidocaine in prolotherapy. Methods: Various concentrations of dextrose and lidocaine were treated in NIH-3T3. Viability was examined with trypan blue exclusion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration assay was performed for measuring the motile activity. Extracellular signal-regulated kinase (Erk) activation and protein expression of collagen I and α-smooth muscle actin (α-SMA) were determined with western blot analysis. Results: The cell viability was decreased in concentrations of more than 5% dextrose and 0.1% lidocaine. However, in the concentrations 1% dextrose (D1) and 0.01% lidocaine (L0.01), fibroblasts proliferated mildly. The ability of migration in fibroblast was increased in the D1, L0.01, and D1 + L0.01 groups sequentially. D1 and L0.01 increased Erk activation and the expression of collagen I and α-SMA and D1 + L0.01 further increased. The inhibition of Erk activation suppressed fibroblast proliferation and the synthesis of collagen I. Conclusions: D1, L0.01, and the combination of D1 and L0.01 induced fibroblast proliferation and increased collagen I synthesis via Erk activation.

쥐 간세포의 일차배양과 분화기능 측정 (Primary culture of adult rat hepatocytes and assay of hepatic functions)

  • 김진희;이재호박정극최태부
    • KSBB Journal
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    • 제7권4호
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    • pp.271-277
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    • 1992
  • 쥐 간세포를collagenase perfusion method에 의해 분리한 뒤 collagen coated dish와floating collagen membrane에서 일차배양하여 간세포의 분화기능을 조사하였다. 두 경우 모두 간세포의 생존율은 5일 이후부터 점차 감소하였거나 간세포에 의한 암모니아 처리기능과 albumin생성기능은 약 7일간 유지되었다. 또 이러한 분화기능에 유지는 ollagen coated dish보다floating collagen membrane이 유리한 것으로 나타났다.

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생간건비탕가삼칠근(生肝健脾湯加三七根)이 흰쥐의 간섬유화 억제에 미치는 영향 (Inhibitory Effect of Saengangeonbitang-gasamchilgn on Liver Fibrosis in Rat)

  • 이은;김영철;고흥
    • 대한한방내과학회지
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    • 제29권2호
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    • pp.500-511
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    • 2008
  • The aim of this study was to investigate the inhibitory effect of Saengangeonbitang-gasamchilgn(SGGBTGSCG) on collagen production in rat hepatic stellate cells(HSC) and on the TAA-induced chronic liver injury model in rats. Methods : 1) HSCs were treated with SGGBTGSCG extract powder(50% EtOH SGGBTGSCG, dw SGGBTGSCG). After the treatment, MTT assay, BrdU assay and procollagen assay were done. In addition, gene expressions of collagen type $1{\alpha}2$, ASMA, TIMP1, and TIMP2, all of which are known to be associated with liver fibrosis, were analyzed by RT-PCR. 2) Liver fibrosis was developed in rats by injection of TAA 3 times a week for 5 weeks. After the SGGBTGSCG-treatment, body weight, liver & spleen weight, liver function test, the complete blood cell count and the change of portal pressure were studied. Results : In MTT assay, SGGBTGSCG significantly decreased the viability of HSCs in a dose-dependent manner. In BrdU assay, SGGBTGSCG significantly inhibited the HSC proliferation in a dose-dependant manner. In procollagen assay, SGGBTGSCG decreased procollagen production by HSC. In the change of rats' liver and spleen weight, TAA+SGGBTGSCG groups showed little difference compared with TAA-only group. In the liver function test, SGGBTGSCG decreased the serum level of ALT, AST, and Alp elevated by TAA. In the complete blood cell count, SGGBTGSCG significantly decreased WBC elevated by TAA and increased RBC and Hct lowered by TAA. In the change of portal pressure, SGGBTGSCG decreased portal pressure elevated by TAA. Conclusions : These results suggest that SGGBTGSCG is beneficial in the treatment of cirrhotic patients as well as for patients with chronic hepatitis.

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Effect of Hijikia fusiforme extracts on degenerative osteoarthritis in vitro and in vivo models

  • Kwon, Han Ol;Lee, Minhee;Kim, Ok-Kyung;Ha, Yejin;Jun, Woojin;Lee, Jeongmin
    • Nutrition Research and Practice
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    • 제10권3호
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    • pp.265-273
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    • 2016
  • BACKGROUND/OBJECTIVES: The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model. MATERIALS/METHODS: In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after $H_2O_2$ ($800{\mu}M$, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs. RESULTS: In the in vitro assay, cell survival after $H_2O_2$ treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and $PGE_2$ were significantly inhibited by treatment with oral administration of HFE extract. CONCLUSION: HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.

Effect of Cinnamomum Cassia on Cartilage Protection in Rabbit and Human Articular Cartilage

  • Baek, Yong-Hyeon;Huh, Jeong-Eun;Lee, Jae-Dong;Choi, Do-Young;Park, Dong-Suk
    • 대한한의학회지
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    • 제28권4호
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    • pp.148-157
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    • 2007
  • Background & Objective: Articular cartilage is a potential target for drugs designed to inhibit the activity of matrix metalloproteinases (MMPs) to stop or slow the destruction of the proteoglycanand collagen in the cartilage extracellular matrix. The purpose of this study was to investigate the effects of Cinnamomum cassia in inhibiting the release of glycosaminoglycan (GAG), the degradation of collagen, and MMP activity in rabbit and human articular cartilage explants. Methods: The cartilage-protective effects of Cinnamomum cassia were evaluated by using glycosaminoglycan degradation assay, collagen degradation assay, colorimetric analysis of MMP activity, measurement of lactate dehydrogenase activity and histological analysis in rabbit cartilage explants culture. Results: Interleukin-1a (IL-1a) rapidly induced GAG, but collagen was much less readily released from cartilage explants. Cinnamomum cassia significantly inhibited GAG and collagen release in a concentration-dependent manner. Cinnamomum cassia dose-dependently inhibited MMP-1, MMP-3 and MMP-13 activities from IL-1a-treated cartilage explants culture when tested at concentrations ranging from 0.02 to 1 mg/ml. Conclusion : These results indicate that Cinnamomum cassia inhibits the degradation of proteoglycan and collagen through the down regulation of MMP-1, MMP-3 and MMP-13 activities of IL-1a-stimulated rabbit and human articular cartilage explants.

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