• Title/Summary/Keyword: Functional assay

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Isolation of Prenylated Isoflavonoids from Cudrania tricuspidata Fruits that Inhibit A2E Photooxidation

  • Uddin, Golam Mezbah;Lee, Hee-Ju;Jeon, Je-Seung;Chung, Dong-Hwa;Kim, Chul-Young
    • Natural Product Sciences
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    • v.17 no.3
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    • pp.206-211
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    • 2011
  • High-performance liquid chromatography coupled to an online $ABTS^+$-based assay (online HPLC-$ABTS^+$) system was used to determine the principal antioxidants in Cudrania tricuspidata fruits. Six prenylated isoflavonoids (1 - 6) were isolated from C. tricuspidata fruits according to the online HPLC-$ABTS^+$ system. The structures of isolated compounds, alpiniumisoflavone (1), 6,8-diprenylorobol (2), 6,8-diprenylgenistein (3), pomiferin (4), 4'-methylalpiniumisoflavone (5), and osajin (6) were identified by their retention time, UV spectra, ESI-MS, and NMR data. Among these compounds, 6,8-diprenylorobol (2) and pomiferin (4) reduced A2E photooxidation in a dose dependent manner.

Anti-inflammation and Anti-aging Effect of Lactic Acid Bacteria Fermented Lycium chinense Extracts (젖산균 발효 구기자(Lycium chinense ) 추출물의 항염증 및 항노화 효능 연구)

  • Han, Dong-Geun;Lee, Ju-Seong;Gal, Gyeok;Park, Nam-Wan;Kim, Hyun-Jeong;An, Bong-Jeon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.2
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    • pp.113-121
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    • 2022
  • In this study, fermented Lycium chinense (L. chinense) extract was used to analyze the efficacy of nitric oxide (NO) and anti-aging inhibition to verify its usefulness as a functional cosmetic material. L. chinense was extracted with hydrothermal and 70% ethanol as a solvent, fermented, concentrated, and freeze-dried to prepare a sample, followed by MTT assay, NO inhibition assay, western blot assay, and UPLC analysis.As a result of the analysis of NO inhibitory efficacy, fermented L. chinense water extract (FLW) and fermented L. chinense 70% ethanol extract (FLE) showed excellent NO inhibitory efficacy of 47.96% and 56.71%, respectively. As a result of measuring the expression patterns of the wrinkle-related proteins MMP-1, and TRPV-1 through Western blot, both FLW and FLE confirmed the inhibitory efficacy of MMP-1 and TRPV-1. Based on the results of the experiment, the fermented L. chinense extract is expected to have a high application value as a functional cosmetic material related anti-inflammatory, and anti-aging.

Phloroglucinol Inhibits the in vitro Differentiation Potential of CD34 Positive Cells into Endothelial Progenitor Cells

  • Kwon, Yi-Hong;Lee, Jun-Hee;Jung, Seok-Yun;Kim, Jae-Won;Lee, Sang-Hun;Lee, Dong-Hyung;Lee, Kyu-Sup;Lee, Boo-Yong;Kwon, Sang-Mo
    • Biomolecules & Therapeutics
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    • v.20 no.2
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    • pp.158-164
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    • 2012
  • Inhibiting the bioactivities of circulating endothelial progenitor cells (EPCs) results in significant inhibition of neovessel formation during tumor angiogenesis. To investigate the potential effect of phloroglucinol as an EPC inhibitor, we performed several in vitro functional assays using $CD34^+$ cells isolated from human umbilical cord blood (HUCB). Although a high treatment dose of phloroglucinol did not show any cell toxicity, it specifically induced the cell death of EPCs under serum free conditions through apoptosis. In the EPC colony-forming assay (EPC-CFA), we observed a significant decreased in the small EPC-CFUs for the phloroglucinol group, implying that phloroglucinol inhibited the early stage of EPC commitment. In addition, in the in vitro expansion assay using $CD34^+$ cells, treatment with phloroglucinol was shown to inhibit endothelial lineage commitment, as demonstrated by the decrease in endothelial surface markers of EPCs including $CD34^+$, $CD34^+/CD133^+$, $CD34^+/CD31^+$ and $CD34^+/CXCR4^+$. This is the first report to demonstrate that phloroglucinol can inhibit the functional bioactivities of EPCs, indicating that phloroglucinol may be used as an EPC inhibitor in the development of biosafe anti-tumor drugs that target tumor angiogenesis.

Anti-stress Effects of Ethanol Extract of Ziziphus jujuba Against Corticosterone-Induced Apoptosis in PC12 Cells

  • Da Hye Song;Yu Jin Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.119-119
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    • 2022
  • The coronavirus disease 2019 (COVID-19) pandemic may be stressful for people. Public health actions, such as social distancing, can make people feel isolated and lonely and can increase stress and anxiety. As a result, there is a growing interest towards various materials to relieve stress. Thus, the present study aimed to investigate the anti-stress effects of ethanol extract of Ziziphus jujuba in PC12 cells treated with corticosterone and its underling mechanisms. Furthermore, the viability of the cells, the apoptosis of the cells, the level of phosphorylation of extracellular signal-regulated kinases (p-ERKs) expression were measured by MTT assay, LDH assay, Hoechst staining assay and western blotting. Our results showed that the extract of Ziziphus jujuba reversed corticosterone-induced damage in PC12 cells, which increased cell viability, decreased LDH release, and attenuated corticosterone-induced apoptosis as compared with the corticosterone-treated group. Therefore, these data suggest that the extract of Ziziphus jujuba could be a good candidate for development as a functional food supplement in the improve the anti-stress effect.

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Analysis of Antioxidant Activity and Cytotoxicity against Human Cancer Cell Lines of Extract from Atractylodes rhizoma fermented with Ganoderma lucidum Mycelium (창출과 영지버섯 균사체 발효 추출물의 항산화 활성 및 인체 암세포주에 대한 세포독성 분석)

  • Park, Mi Hye;Kim, Mee Ra
    • The Korean Journal of Food And Nutrition
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    • v.30 no.3
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    • pp.454-463
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    • 2017
  • Ganoderma lucidum has been traditionally used as a medicine for treatment of bronchitis, arthritis, and high blood pressure, and it has been reported to display many biological activities including anticancer and immune activities. Since mushroom mycelium is known to have excellent biological activities together with mushroom fruiting body, studies on biological activities of mushroom mycelium have been actively conducted. Thus, the present study compared the biological activities before and after the cultivation of Ganoderma lucidum mycelium on Atractylodes rhizoma. When the radical scavenging activity was assessed by the DPPH assay, ARGL (ethanol extract of Atractylodes rhizoma mycelium fermented with Ganoderma lucidum) showed radical scavenging activity of 5.58~82.56% at concentrations of $10{\sim}500{\mu}g/assay$, while AR (ethanol extract of Atractylodes rhizoma) showed radical scavenging activity of 5.27~72.08% at the same concentrations. When measured by using the ABTS assay, ARGL showed higher radical scavenging activity than AR, which was consistent with the result obtained by the DPPH assay. In the MTT assay, the cytotoxicity of ARGL against all cell lines was higher than that of AR. In particular, the cytotoxicities of AR and ARGL against Hep3B at a concentration of $400{\mu}g/assay$ were 71.81% and 86.40%, respectively. In addition, the result obtained by the SRB assay was consistent with the result obtained by the MTT assay. According to the results mentioned above, there is a high probability that medicinal herb cultures using mycelium can be used as sources of functional foods since the cytotoxicities against cancer cells and antioxidant activities increased when the mycelium was fermented with Atractylodes rhizoma.

Ginsenoside Rh2 attenuates microglial activation against toxoplasmic encephalitis via TLR4/NF-κB signaling pathway

  • Xu, Xiang;Jin, Lan;Jiang, Tong;Lu, Ying;Aosai, Fumie;Piao, Hu-Nan;Xu, Guang-Hua;Jin, Cheng-Hua;Jin, Xue-Jun;Ma, Juan;Piao, Lian-Xun
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.704-716
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    • 2020
  • Background: Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. Methods: The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. Results and conclusion: The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an antieT. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.

Polarity affects the antioxidant and antimicrobial activities of jellyfish (Acromitus hardenbergi) extracts

  • Khong, Nicholas M.H.;Foo, Su Chern;Yau, Sook Kun;Chan, Kim Wei;Yusoff, Fatimah Md.
    • Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.189-201
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    • 2022
  • Jellyfish is an emerging aquaculture species, farmed for Oriental cuisines and nutraceutical ingredients. This study aimed to examine antioxidative and antimicrobial potentials of various fractions of the jellyfish, Acromitus hardenbergi. The bell and oral arms of the jellyfish were sequentially extracted with petroleum ether (PE), dichloromethane (DCM), chloroform (CHCl3), methanol (MeOH), and water (H2O) to extract its bioactive in an increasing polarity gradient. Test fractions were assayed for antiradical activities using electron spin resonance spectrometry, β-carotene-linoleate model and Folin-Ciocalteu assay; and antimicrobial activity against 2 Gram-negative bacteria, 4 Gram-positive bacteria and 2 fungal species using the disc diffusion assay. All fractions were also subjected to Fourier Transform Infrared (FTIR) analysis to identify types of functional groups present. It was found that the hydrophilic extracts (H2O fractions) possessed the most effective radical scavenging activity (p < 0.05) while the lipophilic extracts (PE fractions) the most active antimicrobial activity, especially against Gram-positive bacteria (p < 0.05). Total oxidation substrates content was found to be highest in the PE fractions of jellyfish bell and oral arms (p < 0.05). FTIR data showed that the H2O and MeOH fractions contains similar functional groups including -OH, -C=O, -N-H and -S=O groups, while the PE, DCM, and CHCl3 fractions, the -CH3, -COOH groups. This study showed that A. hardenbergi contains antioxidants and antimicrobials, thereby supporting the traditional claim of the jellyfish as an anti-aging and health-promoting functional food. Bioassay-guided fractionation approach serves as a critical milestone for the strategic screening, purification, and elucidation of therapeutically significant actives from jellyfish.

Study on Anti-Skin Aging Effect of Sanguisorba officinalis L. (지유(地楡)의 피부 노화(老化)에 대한 연구(硏究))

  • Kim, Kyoung-Shin;Tak, Dong-Yul;Kim, Byoung-Soo;Kang, Jung Soo
    • Journal of Haehwa Medicine
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    • v.21 no.2
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    • pp.63-72
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    • 2013
  • To develop a new anti-skin aging cosmetics or functional foods by using antioxidative activity and collagenase inhibitor, a potent collagenase or elastase inhibitor was screened from various extracts of medicinal plants and its optimal extraction condition was investigated. And antioxidative activity, antimicrobial activity and inhibitory of effect against collagenase activity were investigated. In the these results, we selected the Sanguisorba (Sanguisorba officinalis L.) that presents a potential biological activities. Sanguisorba which is very rich in triterpenoid saponin and tannins was recently reported its anti-oxidant activities and phytoestogenic activities in vivo test and many clinical studies. The experiments were carried out in vitro to determine anti-oxidant activities of Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, Elastase and collagenase activity assay. It show that the Sanguisorba extracts have the most significant anti-oxidant on free radical scavenging activity assay, and also inhibited significantly activities of elastase, collagenase. Further, Sanguisorba extracts are activated Type I collagen protein expression in CCD-986sk cells. These result suggest that the Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, elastase and collagenase activity assay, Type I collagen protein expression in CCD-986sk cells effected could be developed cosmetic ingredients for anti-aging.

Establishment of an Assay for P2X7 Receptor-Mediated Cell Death

  • Lee, Song-Yi;Jo, Sooyeon;Lee, Ga Eun;Jeong, Lak Shin;Kim, Yong-Chul;Park, Chul-Seung
    • Molecules and Cells
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    • v.22 no.2
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    • pp.198-202
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    • 2006
  • The $P2X_7$ receptor, an ATP-gated cation channel, induces cell death in immune cells and is involved in neurodegenerative diseases. Although the receptor plays various roles in these diseases, the cellular mechanisms involved are poorly understood and antagonists are limited. Here, the development of a cell-based assay for human $P2X_7$ receptor is reported. We established permanent lines of HEK 293 cells expressing a high level of $hP2X_7$ receptor. Functional activity of the $hP2X_7$ receptor was confirmed by whole-cell patch recording of ATP-induced ion currents. Prolonged exposure to ATP resulted in death of the $hP2X_7$-expressing HEK 293 cells and this cell death could be quantified. Two known $P2X_7$ antagonists, PPADS and KN-62, blocked ATP-induced death in a concentration-dependent manner. Thus, this assay can be used to screen for new antagonists of $hP2X_7$ receptors.