• Title/Summary/Keyword: Collagenase-1 inhibitory activity

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Thiol-dependent Redox Mechanisms in the Modification of ATP-Sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Na-Ri;Cuong, Dang-Van;Kim, Chung-Hui;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.15-23
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    • 2003
  • Cellular redox state is known to be perturbed during ischemia and that $Ca^{2+}$ and $K^2$ channels have been shown to have functional thiol groups. In this study, the properties of thiol redox modulation of the ATP-sensitive $K^2$ ($K_{ATP}$) channel were examined in rabbit ventricular myocytes. Rabbit ventricular myocytes were isolated using a Langendorff column for coronary perfusion and collagenase. Single-channel currents were measured in excised membrane patch configuration of patch-clamp technique. The thiol oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB) inhibited the channel activity, and the inhibitory effect of DTNB was reversed by dithiothreitol (disulfide reducing agent; DTT). DTT itself did not have any effect on the channel activity. However, in the patches excised from the metabolically compromised cells, DTT increased the channel activity. DTT had no effect on the inhibitory action by ATP, showing that thiol oxidation was not involved in the blocking mechanism of ATP. There were no statistical difference in the single channel conductance for the oxidized and reduced states of the channel. Analysis of the open and closed time distributions showed that DTNB had no effect on open and closed time distributions shorter than 4 ms. On the other hand, DTNB decreased the life time of bursts and increased the interburst interval. N-ethylmaleimide (NEM), a substance that reacts with thiol groups of cystein residues in proteins, induced irreversible closure of the channel. The thiol oxidizing agents (DTNB, NEM) inhibited of the $K_{ATP}$ channel only, when added to the cytoplasmic side. The results suggested that metabolism-induced changes in the thiol redox can also modulate $K_{ATP}$ channel activity and that a modulatory site of thiol redox may be located on the cytoplasmic side of the $K_{ATP}$ channel in rabbit ventricular myocytes.

Change of Antioxidative Activity at Different Harvest Time and Improvement of Atopic Dermatitis Effects for Persimmon Leaf Extract (감잎(柿葉) 추출물의 채취시기에 따른 항산화 활성 변화 및 아토피성 피부염 개선효과)

  • Jung, Woo-Young;Jeong, Jong-Moon
    • The Korea Journal of Herbology
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    • v.27 no.1
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    • pp.41-49
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    • 2012
  • Objectives : The purpose of this study was to investigate the anti-oxidative and anti-atopic dermatitis effects of persimmon leaf extract obtained from Cheongdo-gun, where more than 60% of Korean persimmon is produced. Methods : Anti-oxidative effects of the crude persimmon leaf extract harvested monthly between May and November were determined by in vitro assay using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide dismutase (SOD)-like reaction. Anti-atopic dermatitis effects of the crude persimmon leaf extract were determined by using collagenase type I inhibition assay and by quantitative assays including serum histamine, prostaglandin E metabolite and leukotriene $B_4$ levels in animal model of atopic dermatitis using Balb/c mice. Results : Persimmon leaf extract harvested in May had higher levels of total phenolic compounds (182.24 mg/g) and flavonoids (23.05 mg/g) than the ones of different month extract. Also, persimmon leaf extract harvested in May showed the most effective extract scavenging activities of DPPH free radical ($13.39{\pm}0.21\;{\mu}g/ml$) and superoxide anion radical ($40.52{\pm}2.32\;{\mu}g/ml$), leading to use the persimmon leaf extract harvested in May for the experiments hereafter. Persimmon leaf extract showed $326.71{\pm}4.6\;{\mu}g/ml$ of 50% inhibitory concentration ($IC_{50}$) for collagenase type I which is responsible for the degradation of extracellular matrix. In addition, persimmon leaf extract application group could decrease serum levels of histamine, prostaglandin E metabolite and leukotriene $B_4$ compared to the negative control in animal model of atopic dermatitis. Especially, persimmon leaf extract showed a significantly decreased serum leukotriene $B_4$ level relative to the levels of histamine and prostaglandin E metabolite. Conclusions : Persimmon leaf extract showed anti-oxidative and anti-atopic dermatitis effects in vitro and in vivo. These results suggest that persimmon leaf extract may have immunoregulatory function for alleviating atopic dermatitis by decreasing collagenase activity and mast cell activation.

The Antioxidant and Anti-aging Effects of Treatment with Schisandra chinensis Seeds Fractions in UVB-irradiated Human HaCaT Cells (자외선 B에 유도된 사람유래 HaCaT cells에서의 오미자 종자 분획물의 항산화 및 항노화 효과)

  • Choi, Eun-Young;Sohn, Ho-Yong;Lee, Jin-Tae
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1071-1079
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    • 2019
  • In this study, we investigated the antioxidant and anti-aging activities of Schisandra chinensis seed fractions by adding them to UVB-irradiated HaCaT cells and analyzing the suppression of matrix metalloproteinases (MMPs). An electron-donating assay, ABTS radical scavenging assay, and hydrogen peroxide scavenging assay showed that the ethyl acetate fraction of S. chinensis seed (SCEAf) has scavenging activities. A collagenase inhibition activity assay showed that SCEAf had inhibitory effects of over 92.3% at $500{\mu}g/ml$ concentration. An MTT assay was performed to determine the cytotoxicity of SCEAf in HaCaT cells and the results showed that SCEAf increased cellular viability in a concentration-dependent manner. In addition, SCEAf was found to increase the viability of cells irradiatged by UVB $50mJ/cm^2$. To examine the anti-aging effects of SCEAf on HaCaT cells irradiated with UVB $50mJ/cm^2$, the expression of MMP-1 and -3 was analyzed by Western blotting and reverse-transcription polymerase chain reaction. The results showed that MMP-1 and -3 proteins and mRNA levels were downregulated in a concentration-dependent manner in response to SCEAf. These results suggest that SCEAf can prevent aging and alleviate aging symptoms by inhibiting collagenase activity in skin keratinocytes damaged by UVB. Therefore, S. chinensis seeds may have the potential for use as functional ingredients with anti-aging effects in the cosmetic and food industries.

Beauty Food Activities of Isolated Phenolic Compounds from Tetragonia tetragonioides (번행초(Tetragonia tetragonioides)로부터 추출한 Phenolic Compounds의 미용 식품 활성)

  • Jo, Jae-Bum;Lee, Eun-Ho;Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.3
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    • pp.333-341
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    • 2016
  • This study examined the beauty food activities of water and ethanol extracts from Tetragonia tetragonioides. Content of phenolic compounds extracted with water and 50% ethanol extracts were 3.29 mg/g and 4.14 mg/g, respectively. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activities of water and ethanol extracts were 98.45% and 91.20%, respectively, at $200{\mu}g/mL$ of phenolics. 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical decolorization activity was 97.28% for water extracts and 97.83% for ethanol extracts at $100{\mu}g/mL$ of phenolics. Antioxidant protection factor (PF) was 1.77 PF for water and ethanol extracts at $200{\mu}g/mL$ of phenolics. Thiobarbituric acid reactive substances of water and ethanol extracts were 94.77% and 95.64%, respectively, at $100{\mu}g/mL$ of phenolics. Tyrosinase inhibitory activity, which is related to skin-whitening, was confirmed to be 34.96% for ethanol extracts at $200{\mu}g/mL$ of phenolics. Elastase inhibitory activity and anti-wrinkle effect of 50% ethanol extracts were 78.9% at $200{\mu}g/mL$ of phenolics. Collagenase inhibitory activity of ethanol extracts was 61.29% at $200{\mu}g/mL$ of phenolics. Astringent effect was not detected in water extracts but was 7.82% for 50% ethanol extracts at $200{\mu}g/mL$ of phenolics. Hyaluronidase inhibitory activity as a measure of anti-inflammation was confirmed to be 81.04% for water extracts at $200{\mu}g/mL$ of phenolics. Based on these results, Tetragonia tetragonioides extracts can be used as a functional material and functional beauty food with antioxidant effects.

In Vitro Cytotoxicity, Skin Regeneration, Anti-wrinkle, Whitening and In Vivo Skin Moisturizing Effects of Oncheongeum (온청음 물 추출물의 세포독성, 피부재생, 주름개선, 미백 및 보습 효과)

  • An, Tteul-E-Bom;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.1
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    • pp.14-34
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    • 2016
  • Objectives: The objective of this study was to evaluate the effects of cytotoxicity, skin regeneration, anti-wrinkle, whitening and skin moisturizing of Oncheongeum (OCE).Methods: The cytotoxicity of OCE lyophilized aqueous extracts (yield=13.82%) was observed against human normal fibroblast cells and B16/F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also evaluated through the assay of collagen type I synthesis compared to the transformation of the growth factor (TGF)-β1, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays compared to oleanolic acid (OA), and elastase inhibitory effects compared to phosphoramidon disodium salt (PP). In addition, OCE’s whitening effects were measured by a tyrosinase inhibitory assay and melanin formation test in B16/F10 murine melanoma cells compared to arbutin, and skin moisturizing effects were observed through a mouse skin water content test, respectively. Results: No OCE treatment-related cytotoxic effects appeared on human normal fibroblasts and B16/F10 murine melanoma cells. OCE concentration-dependently increased the collagen Type I synthesis on human normal fibroblast cells, and also effectively inhibited hyaluronidase, elastase, collagenase and MMP-1 activities. In addition, OCE inhibited melanin production of B16/F10 murine melanoma cells and activity of tyrosinase. And significant and dose-dependent increases of skin water content were detected in OCE-treated mouse skin compared to vehicle control skins. Conclusions: OCE showed favorable and sufficient effects in skin regeneration, anti-wrinkle, whitening and skin moisturizing in this experiment. But more detail mechanisms and studies on the skin protective efficiency of in vivo are needed with the screening of active biological compounds in individual OCE herbs.

Mechanisms of Suppression of Matrix Metalloproteinases in UVB-Irradiated HaCaT Keratinocytes of Colored Rice Varieties (UVB에 조사된 HaCaT Keratinocytes에서의 유색미에 의한 Matrix Metalloproteinases 발현억제 메커니즘)

  • Choi, Eun-Young;Lee, Jae-Bong;Kim, Do-Hoon;Kwon, Yong-Sham;Cheon, Jung-Yoon;Lee, Jin-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.562-571
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    • 2017
  • In this study, we investigated the anti-oxidant activities [electron-donating ability (EDA), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, and reactive oxygen species (ROS) inhibitory activity], anti-wrinkle activities [collagenase inhibitory activity, suppression and/or phosphorylation of matrix metalloproteinases (MMPs), and mitogen-activated protein (MAP) kinase activity], and mRNA expression levels using reverse transcription-polymerase chain reaction (RT-PCR) assay in ultraviolet (UV) B ray ($50mJ/cm^2$)-irradiated human keratinocyte HaCaT cells. Josaengheugchal, Sinneungheugchal (SE), Shintoheug rice, Heugjinju rice, and Heugseol (HE) among colored rice varieties were reported to have excellent antioxidant properties. In the EDA and ABTS radical scavenging assays, extracts of the five colored rice varieties had scavenging activities of 72% at concentrations higher $50{\mu}g/mL$. In the collagenase inhibition assay, ethanol extracts of the five colored rice varieties showed high inhibitory effects of about 60% at concentrations higher $25{\mu}g/mL$. In the ROS inhibition assay, ethanol extracts of HE and SE showed very excellent inhibition efficacies at all concentrations. We determined molecular biological mechanisms of MMPs (MMP-1, -3, -8, and -13) and mitogen-activated protein kinase (MAPK) with HE, and the results show that HE suppressed expression of MMPs and phosphorylation of MAPK and increased expression of pro-collagen type I in UVB-irradiated cells. It was also confirmed by RT-PCR that HE reduced expression of MMPs mRNA. Therefore, these results suggest that HE has anti-wrinkle and collagen production effects and may be used as a material in the development of functional food and cosmetic industries.

Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

Biological Activity of Fermented Gryllus bimaculatus extracts (발효 쌍별귀뚜라미 추출물의 생리활성 연구)

  • Park, In-Sun;Lim, Hyeon-Ji;Jeong, Seung-Il;Jung, Chan-Hun
    • Korean Journal of Pharmacognosy
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    • v.52 no.2
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    • pp.92-98
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    • 2021
  • Gryllus bimaculatus extract (GbE) have reported that anti-inflammatory activity by suppression of pro-inflammatory cytokines. However, the effects of fermented Gryllus bimaculatus extract (FGbE) have not yet been investigated. In this study, we evaluated the anti-inflammatory and anti-wrinkle effect of the fermented Gryllus bimaculatus extracts using Bacillus subtilis (JB PMB-18) in RAW264.7 cells. Both GbE and FGbE exerted no cytotoxic effects until 1000 ㎍/mL concentration. FGbE decreased NO production and decreased iNOS and COX-2 mRNA levels in a concentration-dependent manner. In addition, the protein production of inflammatory cytokines TNF-α, IL-1β and IL-6 was effectively reduced compared to the GbE. Inhibitory activities of elastase and collagenase associated with skin wrinkle improvement were measured to be 45% and 69%, respectively, at a concentration of 500 ㎍/mL in FGbE. From these results, FGbE can be used as a health functional food and skin functional cosmetic materials for preventing inflammatory diseases because it has excellent anti-inflammatory and anti-wrinkle effects.

Effects of Dendrobii herba and Punica granatum Extract on the Anti-oxidant, Anti-inflammatory, Anti-wrinkle and Whitening (석곡(石斛), 석류(石榴)의 항산화, 항염증, 주름, 미백에 미치는 영향)

  • HwangBo, Min;Roh, Seok-Sun;Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.3
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    • pp.11-32
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    • 2010
  • Objective : The aim of this study is to determine the effects of Dendrobii herba extract and Punica granatum extract on skin disease and skin beauty. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH radical, xanthine oxidase activity. In the next experiment, to investigate anti-inflammatory activity assay, examined by relations in NO synthesis, IL-$1{\beta}$, IL-6, TNF-${\alpha}$, NF-${\kappa}B$, COX-2, MAP kinase. To study Skin wrinkle formation effect, we were examined by tyrosinase activities, melanin synthesis in MNT-1 cell. Results : 1. In an anti-oxidant test, Dendrobii and Punica granatum extract showed high radical scavenging activity. 2. In an anti-inflammatory test, Dendrobii herba and Punica granatum extract weakly inhibited the lipopolysaccharide(LPS)-induced nitric oxide(NO) release from RAW 246.7 macrophage cells. Dendrobii herba and Punica granatum extract also inhibited LPS-induced IL-$1{\beta}$ and COX-2 expressions. The inhibitory effect of Dendrobii herba and Punica granatum extract on macrophage activation were via the inhibition of NF-${\kappa}B$, evidenced by transient transfection assay. however, Dendrobii herba and Punica granatum extract did not have any effects about activation of Jun-N-terminal kinase(JNK) and inhibition of p38 MAP kinase in RAW 264.7 cells. 3. In the skin wrinkle formation assay, Dendrobii herba and Punica granatum extract weakly inhibited collagenase and elastase, however it was not statistically significant. 4. In the skin whitening assay, Dendrobii herba and Punica granatum extract weakly inhibited tyrosinase activity, however, it was not statistically significant. They did not have any effect on melanin synthesis, indicating that they could not be applicable for skin whitening. Conclusion : Dendrobii herba extract and Punica granatum extract may play a significant role in skin disease and skin beauty.

Biological Activities of Extracts from Okkwang (Castanea crenata) Chestnut Bur (옥광(Castanea crenata) 밤송이 추출물의 생리활성 효과)

  • Lee, Eun Ho;Hong, Shin Hyub;Cho, Young Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.572-580
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    • 2017
  • The contents of phenolic compounds in water and 40% ethanol extracts from Okkwang (Castanea crenata) chestnut bur solid (OCS) were $11.24{\mu}g/50{\mu}g$ solid and $10.28{\mu}g/50{\mu}g$ solid, respectively. The 1,1-diphenyl-2-picrylhydrazyl free radical scavenging and 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulfonic acid) radical decolorization activities of water and ethanol extracts were 85% and 100% as well as 87% and 86% at a solid content of $50{\mu}g/mL$, respectively. The anti-oxidant protection factors (PFs) of water and ethanol extracts at a solid content of $200{\mu}g/mL$ were 1.22 PF and 1.45 PF, respectively. Thiobarbituric acid reactive substance were 83% in water extract and 73% in ethanol extract at a solid content of $200{\mu}g/mL$. The inhibitory activities against xanthine oxidase in water and ethanol extracts were 54% and 43% at a solid content of $200{\mu}g/mL$, respectively. The inhibitory activities against ${\alpha}$-glucosidase were 95% in water extract and 96% in ethanol extract at a solid content of $50{\mu}g/mL$. Tyrosinase inhibitory activity was 27% in ethanol extract at a solid content of $200{\mu}g/mL$. The collagenase and elastase inhibitory activities as anti-wrinkle effect were 93% and 11% in water extract as well as 94% and 56% in ethanol extract at a solid content of $200{\mu}g/mL$. Hyaluronidase inhibitory activity as anti-inflammatory effect of water and ethanol extracts were 96% and 52% at a solid content of $200{\mu}g/mL$, respectively. The results show that extracts from OCS can be used as a functional resource with antioxidant, anti-gout, carbohydrate degradation inhibitory, whitening, anti-wrinkle, and anti-inflammatory activities.