• YAKHAK HOEJI
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• v.48 no.2
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• pp.165-170
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• 2004

The production of matrix metalloproteinases (MMPs) by the UV irradiated skin fibroblast and the degradation of extracellular matrix (ECM) by these enzymes is known as one of the main reasons of photoaging. In this paper, to investigate the relationship between aging and Selaginella tamariscina extract (STE), we investigated the effects of antioxidant and expression of UVA-induced MMP-1 in human dermal fibroblasts. STE was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$/ values of 65.1 $\mu\textrm{g}$/$m\ell$ against 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical and 40.9 $\mu\textrm{g}$/$m\ell$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. UVA induced MMP expression was reduced 75.5% by treatment with STE, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore STE was able to significantly inhibition of MMP expression in protein and mRNA level. All these results suggested that STE may act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.243-249
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• 2007

The French paradox has been attributed to the antioxidant properties of flavonoids present in the red wine. Amentoflavone(AF) is a bi-flavonoid compound with anti-fungal and anti-inflammatory activities. We isolated AF from Selaginella tamariscina, and studied its effects on nuclear factor-B(NF-B)-mediated MMP-9 gene expression in RAW264.7 cells. AF blocked the lipopolysaccharide(LPS)-induced expression of MMP-9. Zymographic and immunoblot analyses showed that AF suppressed LPS-induced MMP-9 expression in a dose-dependent manner. To clarify the mechanistic basis for its inhibition of MMP-9 induction, we examined the effect of AF on the transactivation of MMP-9 gene by luciferase reporter activity using -1.59 kb flanking region. AF potently suppressed the reporter gene activity. This inhibition was characterized by down-regulation of MMP-9, which was transcriptionally regulated at NF-B site and activation protein-1 (AP-1) site in the MMP-9 promoter, two important nuclear transcription factors that are involved in MMP-9 expression. These findings indicate the efficacy of AF in inhibiting MMP-9 expression through the transcription factors NF-B and AP-1 on LPS-induced RAW264.7 cells.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.10a
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• pp.81-90
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• 2004

Sialic acid containing glycosphingolipids (gangliosides) have been implicated in the regulation of various biological phenomena such as atherosclerosis. Recent report suggeststhat exogenously supplied disialoganglioside (GD3) serves a dual role in vascular smooth muscle cells (VSMC) proliferation and apoptosis. However, the role of the GD3 synthase gene in VSMC responses has not yet been elucidated. To determine whether a ganglioside is able to modulate VSMC growth. the effect of overexpression of the GD3 synthase gene on DNA synthesis was examined. The results show that the overexpression of this gene has a potent inhibitory effect on DNA synthesis and ERK phosphorylation in cultured VSMC in the presence of PDGF. The suppression of the GD3 synthase gene was correlated with the down-regulation of cyclinE/CDK2. the up-regulation of the CDK inhibitor p21 and blocking of the p27 inhibition,whereas up-regulation of p53 as the result of GD3 synthase gene expression was not observed. Consistently, blockade of GD3 function with anti-GD3 antibody reversed VSMC proliferation and cell cycle proteins. The expression of the CD3 synthase gene also led to the inhibition of TNF--induced matrix metalloproteinase-9 (MMP-9) expression in VSMC as determined by zymography and immunoblot. Furthermore, GD3 synthase gene expression strongly decreased MMP-9 promoteractivlty in response to TNF-. This inhibition was characterized by the down-regulation of MMP-9,which was Iranscriptionally regulated at NF-B and activation protein-1 (AP-1) sites in the MMP-9promoter Finally, the overexpression of MMP-9 in GD3 synthase transfectant cells rescued VSMC proliferation. However MMP-2 overexpression was not affected the cell proliferation. These findings suggest that the fl13 synthase gene represents a physiological modulator of VSMC responses that may contribute to plaque instability in atherosclerosis.

• BMB Reports
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• v.32 no.1
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• pp.60-66
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• 1999

Matrix metalloproteinase-2 (MMP-2; 72-kDa gelatinase; 72-kDa type IV collagenase; gelatinase A) plays an important role in normal physiological processes and in many pathologic processes such as arthritis and metastasis of cancer. Tissue inhibitor of metalloproteinases-2 (TIMP-2) binds to proMMP-2 or mature MMP-2 at a 1:1 ratio and inhibits the catalytic activity of MMP-2. We demonstrated that the baculovirus/insect cell system does not have TIMP-2 activity. The human proMMP-2 free of TIMP-2 was expressed in the expression system and purified by one-step affinity chromatography using gelatin-Sepharose. The free proMMP-2 was autoactivated to the mature MMP-2 and autodegraded into smaller molecular weight forms in the absence of external activator. The activation and autodegradation of the proMMP-2 was much more rapid in the presence of 4-aminophenylmercuric acetate (APMA). Addition of TIMP-2 inhibits both APMA-induced activation and autodegradation of the free proMMP-2. However, an increasing concentration of TIMP-2 more readily inhibited activation of the free proMMP-2 than autodegradation. These results demonstrate that TIMP-2 plays roles in inhibition of both activation and autodegradation of the free proMMP-2 in addition to inhibition of the catalytic activity of MMP-2.

• The Korea Journal of Herbology
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• v.31 no.4
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• pp.53-60
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• 2016

Objective : The purpose of this study was to investigate anti-aging and antioxidant effects of extracts of Nelumbo nucifera leaves (NN-L) using ethanol on skin .Methods : Each part of leaves(NN-L), flowers(NN-F) and stem(NN-S) was extracted with 70% ethanol. We performed radical scavenging assay(DPPH, ABTS+, Superoxide anion radical), elastase inhibition assay, collagenase inhibition assay. NN-L extracts were tested for cell viability(MTT assay), MMP-1 inhibition and MMP-1 protein expression on CCD-986sk cells (human fibroblast line).Results : 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. We measured its free radical scavenging activity, elastase inhibitory activity and expression of MMP-1 (matrix metalloprotease-1) in human fibroblast cells. Among the parts of Nelumbo nucifera, NN-L showed the highest antioxidant activities and in radical scavenging. DPPH, ABTS+ and Superoxide anion radical scavenging activity of NN-L at concentration of 1,000 μg/mL were 91.43%, 99.31% and 73.7% respectively. In vitro elastase and collagenase inhibition effects of NN-L at concentration of 1,000 μg/mL was 42.8% and 55.3% respectively. The ethanol extract of NN-L showed cell viability of 95.4% in 50 μg/mL concentration. In addition, The results from Western blot assay showed that NN-L decreased the expression of MMP-1 protein in a dose-dependent manner (by up to 35.0% at 50 μM).Conclusion : The findings suggest that the NN-L great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

• Advances in Traditional Medicine
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• v.5 no.4
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• pp.283-293
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• 2005

Our previous study showed that a novel synthetic shikonin derivative, 6-(1-hydroxyimino-4-methylpentyl)5,8-dimethyoxy 1,4-naphthoquinone S-52 (DMNQ S-52) induced apoptosis. In the present study, we investigated its anti-metastatic activities as compared with shikonin because DMNQ S-52 was synthesized for overcoming weak points of shikonin such as high toxicity, low solubility and deleterious effects. DMNQ S-52 showed the weaker cytotoxicity $(IC_{50};\;12.3{\pm}1.6\;{\mu}M)$ against Lewis lung carcinoma (LLC) cells than that of shikonin $(IC_{50};\;4.2{\pm}1.1\;{\mu}M)$. DMNQ S-52, at non-toxic concentrations $(less\;than\;10\;{\mu}M)$, significantly inhibited the invasion and migration of LLC cells. DMNQ S-52 also significantly inhibited the production of MMP-9, MTl-MMP and uPAR. Moreover, daily i.p. administration of DMNQ S-52 at dose of 5 mg/kg in mice resulted in a potent inhibition of the primary tumor size of LLC in the lung as well as the metastasis of lymph nodes. These findings suggest that the DMNQ S-52 has therapeutic potential to inhibit metastasis via inhibition of MMP family and uPA/plasminogen system.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.2 s.51
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• pp.161-167
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• 2005

We investigated the effect on inhibition of matrix metalloproteinase (MMP) in human dermal fibroblast (HDF) by production of exopolysaccharide (GF-glucan) from mycelial culture of Grifola frondosa HB0071. The photoprotective potential of GF-glucan was tested in HDF exposed to ultraviolet-A (UVA) light. It was revealed that GF-glucan had an inhibitory effect on MMP-1 expression in UVA-irradiated HDF without any significant cytotoxicity. The treatment of UVA-irradiated HDF with GF-glucan resulted in a dose-dependent degrease in the expression level of MMP-1 protein and mRNA (by maximum $54.4\%$ at an $0.5\%$ GF-glucan). These results suggest that GF-glucan obtained from mycelial culture of G. frondosa HB0071 may contribute to inhibitory action in photoaging by reducing the MMP-1 related matrix degradation system.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.3 s.47
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• pp.393-397
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• 2004

L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

• 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.

• Journal of Life Science
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• v.23 no.10
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• pp.1239-1245
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• 2013

The purpose of this study was to research the biological activity of ethanol extract from Smilax china L. which is a vine shrub belonging to the lily family. For antiwrinkle effects, elastase inhibition effect of ethanol and water extracts from S. china L. showed 41.1% and 16.3% at $1,000{\mu}g/ml$ concentration. The collagenase inhibition effect of ethanol and water extracts from S. china L. showed more than 96.6% and 60.0% at $1,000{\mu}g/ml$ concentration. As a result of having fibroblast measured cell viability on fibroblast cell of ethanol extract from S. china L., it showed 71.7% with cell viability at $100{\mu}g/ml$ concentration. At $50{\mu}g/ml$ concentration, the procollagen biosynthesis effect of ethanol extract from S. china L. was 139.86%. At the same concentration, the matrix metalloprotease (MMP)-1 inhibition effect of the ethanol extract was 74.9%. According to the results of Western blot of ethanol extract from S. china L., the expression of the MMP-1 protein was decreased by 35% at $50{\mu}g/ml$ concentration. Reverse transcription-polymerase chain reaction (PCR) of ethanol extract from S. china L. showed that the expression of MMP-1 mRNA was decreased by 45% at $50{\mu}g/ml$ concentration. The findings suggest that 70% ethanol extract from S. china L. (SC) has great potential as a cosmeceutical ingredient with antiwrinkle effects.