• Title/Summary/Keyword: collagenase activity

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Beauty food activities of wild-cultivated Ginseng (Panax ginseng C.A. Meyer) ground part (산양삼(Panax ginseng C.A. Meyer) 지상부위의 미용 식품 활성)

  • Kim, Myeong-Wook;Lee, Eun-Ho;Kim, Ye-Jin;Park, Tae-Soon;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.33-38
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    • 2018
  • This study aimed to investigate the beauty food activities of wild-cultivated ginseng (Panax ginseng C.A. Meyer). wild-cultivated ginseng extracts were analyzed for antioxidant, skin whitening, anti-wrinkle effect was measured in water and 70% ethanol extract. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical decolorization activities of water and 70% ethanol extracts were 16.69 and 2.18% as well as 4.04 and 3.25% at a solid content of $200{\mu}g/mL$, respectively. The antioxidant protection factors (PF) of water and 70% ethanol extracts at a solid content of $200{\mu}g/mL$ were 1.06 PF and 1.09 PF, respectively. Thiobarbituric acid reactive substance (TBARs) were both 96% at a solid content of $200{\mu}g/mL$. As PF and TBARs showed higher activity than DPPH and ABTS, we could know that antioxidant activity in the lipophilic component of wood-cultivated ginseng were superior to water-soluble component of wood-cultivated ginseng. Tyrosinase inhibitory activity was 10.97 and 52.39% in water and 70% ethanol extracts at a solid content of $200{\mu}g/mL$. The collagenase and elastase inhibitory activities as anti-wrinkle effect were 15.71 and 20.43% in water extracts as well as 32.26 and 86.74% in 70% ethanol extract at a solid content of $200{\mu}g/mL$. The results show that anti-wrinkle effect was the best among the other experiments. This extracts from wood-cultivated ginseng, therefore, seems to be a potent beauty food resource against wrinkles.

A Study of Anti-wrinkle Activities as a Functional Cosmetic Ingredient of Rhododendron brachycarpum Extracts (만병초(Rhododendron brachycarpum) 추출물의 기능성 화장품 소재로써의 주름개선 활성에 관한 연구)

  • Yeom, Hyeon-Ji;Oh, Min-Jeong;Chae, Jung-Woo;Lee, Jin-Young
    • Journal of Life Science
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    • v.32 no.8
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    • pp.622-632
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    • 2022
  • The purpose of this study was to investigate activities as functional cosmetic materials, focusing on Rhododendron brachycarpum (RB) and Rhododendron fortunei (RF). The tyrosinase inhibitory effect, related to skin-whitening, was 32.6% and 39.3% respectively at the concentration of 1,000 ㎍/ml. The elastase inhibitory effect, related to skin anti-wrinkling activity, was 30% and 36.2% at 1,000 ㎍/ml concentration. Collagenase inhibitory activity showed 77.7%, and 80.2% respectively at 1,000 ㎍/ml concentration, which demonstrated excellent inhibitory activity. For a cell viability test, that measured on fibroblast cells by RB and RF extracts. Furthermore, the cell viability test showed 100.9% and 98.9% with cell viability at 100 ㎍/ml concentration in CCD-986Sk. The protein expression inhibitory effect of RB and RF extracts was measured by western blot at 25, 50, and 100 ㎍/ml concentrations, and the β-actin was used as a positive control. As a result, western blot of RB and RF extracts was measured by the expression inhibition rate of the MMP-1, MMP-2, MMP-3 protein, and was decreased by 67.2%, 65.5%, 13.6%, and 89.1%, 85.0%, and 62.7% at 100 ㎍/ml concentration. The mRNA expression inhibitory effect of RB and RF extracts was measured by RT-PCR at 25, 50, and 100 ㎍/ml concentrations, and the GAPDH was used as a positive control. According to the results of RT-PCR of RB and RF extracts, the expression inhibition rate of the MMP-1, MMP-2, and MMP-3 mRNA was decreased by 70.1%, 9.1%, 37.9%, and 38.2%, 8.3%, 57.3% at 100 ㎍/ml concentrations. So, RB and RF extracts showed the anti-wrinkle effectiveness as a functional cosmetic material.

Various Physiological and Anti-inflammatory Effects of Sanguisorba officinalis L. Roots as a Functional Cosmetic Material (기능성 화장품 소재로써 오이풀 뿌리(Sanguisorba officinalis L. roots)의 다양한 생리 활성 및 항염증 효과)

  • Seung-Mi Park;Min-Jeong Oh;Hyeon-Ji Yeom;Mi-Ock Shim;Jin-Young Lee
    • Journal of Life Science
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    • v.33 no.5
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    • pp.406-413
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    • 2023
  • In this study, the various physiological and anti-inflammatory activities of Sanguisorba officinalis L. roots (SR) were assessed for potential use as functional cosmetic materials. As a result of measuring electron-donating abilities to determine the antioxidant ability of SR extract, activity increased as the concentration increased, showing an excellent antioxidant capacity of 93.8% at a 1,000 ㎍/ml concentration. Further, the antioxidant power of SR extract, which was determined using an ABTS+ assay measurement, was more than 99% at concentrations of 50 ㎍/ml or more, while the tyrosinase inhibition rate was 37.7% at the highest concentration of 1,000 ㎍/ml. Consequently, the elastase and collagenase inhibition of SR extract measured 84.9% and 90.3%, respectively, at a 1,000-㎍/ml concentration. As a result of confirming the survival rate of Raw 264.7 cells, the cell survival rate was determined to be 80% or more below a 100 ㎍/ml concentration, and subsequent cell-related experiments were conducted at concentrations below 100 ㎍/ml. Furthermore, after applying a NO assay to identify anti-inflammatory activity, it was confirmed that SR extract had an inhibitory rate of 50.8% at a concentration of 500 ㎍/ml, and it was excellent at suppressing the inflammatory expression. As a result of verifying protein expression by treating SR extract in Raw 264.7 cells, it was confirmed that expression was inhibited concentrated in all factors. Therefore, it is judged that SR can be used as a functional cosmetic material with antioxidant, whitening, and wrinkle-improving physiological effects and anti-inflammatory activities.

Isolation and Characterization of Antioxidative Peptides from Enzymatic Hydrolysates of Yellowfin Sole Skin Gelatin (가자미피 젤라틴 가수분해물로부터 항산화성 펩티드의 분리${\cdot}$정제 및 특성)

  • KIM Se-Kwon;LEE Hyun-Chel;BYUN He-Guk;JEON Yon-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.2
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    • pp.246-255
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    • 1996
  • To develop a natural antioxidative peptide, the gelatin was extracted from fish (Yellowfin sole) skin by hot $water(50^{\circ}C)$ extraction method and hydrolyzed with Alcalase, pronase and collagenase through a continuous 3-step membrane reactor. Each step enzymatic hydrolysates were determined the antioxidative activity and their synergistic effects, compared with $\alpha-tocopherol$ and butylated hydroxytoluene (BHT). Also, we tried to investigate the antioxidative disposition of peptide which was successfully separated by gel filtration, ion-exchange chromatography, and HPIC in cultured rat hepatocytes intoxicated with tert-butyl hydroperoxide (TBHP). Second step enzymatic hydrolysate (SSEH) among all hydrolysates and $\alpha-tocoperol$ was showed the strongest antioxidative activity. The optimum concentration of antioxidative activity for SSEH was $1\%(w/w)$ in linoleic acid. The synergistic effects were increased in using the hydrolysate with tocopherol and BHT. In the presence of the peptide isolated from SSEH, supplemented hepatocytes exposed to TBHP showed that delayed cell killing and decreased significantly the lipid peroxidation, compared with hepatocytes not cultured with isolated peptide.

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Testosterone Relaxes Rabbit Seminal Vesicle by Calcium Channel Inhibition

  • Kim, Jong-Kok;Han, Woo-Ha;Lee, Moo-Yeol;Myung, Soon-Chul;Kim, Sae-Chul;Kim, Min-Ky
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.2
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    • pp.73-77
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    • 2008
  • Recent studies have documented that testosterone relaxes several smooth muscles by modulating $K^+$ channel activities. Smooth muscles of seminal vesicles playa fundamental role in ejaculation, which might involve testosterone. This study was aimed to assess the role of testosterone in seminal vesicular motility by studying its effects on contractile agents and on the ion channels of single vesicular myocytes in a rabbit model. The contractile responses of circular smooth muscle strips of rabbit seminal vesicles to norepinephrine ($10{\mu}M$), a high concentration of KCI (70 mM), and testosterone ($10{\mu}M$) were observed. Single vesicular myocytes of rabbit were isolated using proteolytic enzymes including collagenase and papain. Inside-out, attached, and whole-cell configurations were examined using the patch clamp technique. The applications of $10{\mu}M$ norepinephrine or 70 mM KCl induced tonic contractions, and $10{\mu}M$ testosterone (pharmacological concentration) evoked dose-dependent relaxations of these precontracted strips. Various $K^+$ channel blockers, such as tetraethylammonium (TEA; $10{\mu}M$), iberiotoxin ($0.1{\mu}M$), 4-aminopyridine (4-AP, $10{\mu}M$), or glibenclamide ($10{\mu}M$) rarely affected these relaxations. Single channel data (of inside-out and attached configurations) of BK channel activity were also hardly affected by testosterone ($10{\mu}M$). On the other hand, however, testosterone reduced L-type $Ca^{2+}$ currents significantly, and found to induce acute relaxation of seminal vesicular smooth muscle and this was mediated, at least in part, by $Ca^{2+}$ current inhibition in rabbit.

Inhibition of Tumor Invasion and Metastasis by Calcium Spirulan(Ca-SP), a Novel Sulfated Polysaccharide Derived from a Blue-Green Alga Spirulina Platensis

  • Saiki, Ikuo;Murata, Jun;Fujii, Hideki;Kato, Toshimitsu
    • Nutritional Sciences
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    • v.7 no.3
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    • pp.144-150
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    • 2004
  • We have investigated the effect of calcium spirulan(Ca-SP) isolated from a blue-green alga Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose and fructose, on invasion of both B16- BL6 melanoma cells, Colon 26 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters in a concentration-dependent manner. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no inhibitory effect on tumor cell migration to fibronectin-coated filters. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel- and laminin-substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin in a concentration-dependent fashion, while the pretreatment of laminin-substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contraset, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP in a dose-dependent manner. Seven intermittent ⅰ.ⅴ. injection of 100$\mu\textrm{g}$ of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung colonization of B16-BL6 melanoma cells in experimental metastasis model, by inhibiting the tumor invasion of basement membrane Matrigel, probably through the prevention of the adhesion and migration of tumor cells to laminin-substrate and of the heparanase activity.

THE EFFECT OF TENSILE FORCE ON DNA AND PROTEIN SYNTHESIS IN BONE CELLS (인장력이 골조직 세포군의 DNA 및 단백합성에 미치는 영향)

  • Kwon, Oh-Sun;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.24 no.4 s.47
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    • pp.933-943
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    • 1994
  • The present study was undertaken to determine the effect of tensile force on DNA and protein biosynthesis in bone cells, and to identify the cell type(s) which primarily respond to external physical force among the heterogenous bone cell populations. As a prerequisite for this study, two bone cell populations which retain fibroblastic and osteoblastic feature were isolated from fetal rat calvaria with sequential enzyme digestion scheme. Tensile force was delivered to each bone cell population by two acrylic resin plates connected with a orthodontic expansion screw during culture period. Rate of DNA and protein synthesis in each bone cell population were assessed by the incorporated radioactivity of $[^3H]-thymidine$ into DNA and $[^3H]-proline$ into fraction of collagenase-digestible protein and noncollagenous protein, respectively. DNA synthesis of osteoblast-like calvarial cell populations was increased significantly by the application of tensile force for 24 hours. In contrast, no alteration in DNA synthesis of fibroblast-like populations could be observed in response to applied force. Tensile force induced the change in protein synthesis of bone cell populations with the same pattern. Total protein and collagen synthesis were increased whithin 24 hours in osteoblast-like populations, but not in fibroblast-like populations by tensile force application. These findings indicate that physical force can affect cellullar activity of the particular cell population, not all cell Populations residing in bone and osteoblasts respond more sensitively than fibroblasts. So osteoblasts can modulate the behavior of other bone cells including osteoclasts by producing several local regulating factors of bone metabolism. In this context, preferential responsiveness of osteoblasts to applied tensile force observed in this study suggests that osteoblasts may play an important role in regulation of physical force-induced remodelling process.

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THE EFFECTS OF HERBAL EXTRACTS ON PRODUCTION AND ACTIVlTY OF INTERLEUKIN 1${\beta}$ (생약추출물이 Intrerleukin-1 ${\beta}$의 생성 및 활성에 미치는 영향)

  • Cho, Ki-Yeong;Lee, Yong-Moo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.386-396
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    • 1995
  • Interieukin 1${\beta}$ is a potent bone resorptive cytokine which mediates soft tissue destruction through the stimulatidn of prostaglandin production and the induction of collagenase. This constellation of activities suggests a role of IL-1${\beta}$ in the pathogenesis of periodontal disease. The purpose of this study was to evaluate the effects of herbal extracts on production and activity of IL-1${\beta}$. When LPS was added to cultured human blood monocytes, the effects of herbal extracts on the production of IL-1${\beta}$ was evaluate by thymocyte stimulation assay. When rHuIL-1${\beta}$ was added to cultured human gingival fibroblasts, the effects of herbal extracts on production of $PGE_2$ was evaluated by ELISA and when it was added to cultured mouse calvaria, the effects on bone resorption was estimated by .$^{45}Ca$-release bone resorption assay. The herbal extracts that had been used in this study were as follows; Asparagi Radix, Schzandrae Fractus, Zizyphi Fractus and Rhois Galla. The following results were obtained from this study. 1. All these extracts effectively inhibited the production of IL-1${\beta}$ on cultured human blood monocytes. 2. All these extracts effectively inibited the production of $PGE_2$ on cultured human gingival fibroblasts. 3. All these extracts did not effectively inhibit the bone resorption induced by rHulL-1${\beta}$ on cultured mouse calvaria.

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Effect of pH on Calcium-Activated Potassium Channels in Pulmonary Arterial Smooth Muscle Cells of the Rabbit

  • Lee, Suk-Ho;Ho, Won-Kyung;Earm, Yung-E
    • The Korean Journal of Physiology
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    • v.25 no.1
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    • pp.17-26
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    • 1991
  • Single smooth muscle cells of the rabbit pulmonary artery were isolated by treatment with collagenase and elastase. Using the patch clamp technique, potassium channel activity was recorded from the inside-out membrane patch. The channel had a sin히e channel conductance of about 360 pS in symmetrical concentration of K on both sides of the patch, 150 mM, and had a linear current-voltage relationship. During the application of 10 mM tetraethylammonium (TEA) to the intracellular membrane surface, the amplitude of single channel current was reduced and very rapid flickering appeared. The open probability $(P_0)$ of this channel was increased by increasing positivity of the potential across the patch membrane, with e-fold increase by 20 mV depolarization, and by increasing the internal $Ca^{2+}$ concentration. These findings are consistent with those of large conductance Ca-activated K channels reported in other tissues. But the shortening of the mean open time by increasing $[Ca^{2+}]_i$, was an unexpected result and one additional closed state which might be arisen from a block of the open channel by Ca binding was suggested. The $P_0-membrane$ potential relationship was modulated by internal pH. Decreasing pH reduced $P_0$. Increasing pH not only increased $P_0$ but also weakened the voltage dependency of the channel opening. The modulation of Ca-activated K channel by pH was thought to be related to the mechanism of regulation of vascular tone by the pH change.

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Effect of Bevacizumab on Human Tenon's Fibroblasts Cultured from Primary and Recurrent Pterygium

  • Park, Young Min;Kim, Chi Dae;Lee, Jong Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.357-363
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    • 2015
  • The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.