• Title/Summary/Keyword: cell-free extract

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Protective effect of Buddha's Temple extract against tert-butyl hydroperoxide stimulation-induced oxidative stress in DF-1 cells

  • Eun Hye Park;Sung-Jo Kim
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1120-1129
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    • 2023
  • Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.

Radical Intermediate Generation and Cell Cycle Arrest by an Aqueous Extract of Thunbergia Laurifolia Linn. in Human Breast Cancer Cells

  • Jetawattana, Suwimol;Boonsirichai, Kanokporn;Charoen, Savapong;Martin, Sean M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4357-4361
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    • 2015
  • Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

Effects of Puerariae Radix extract on the activity of antioxidant (갈근(葛根) 추출물이 항산화에 미치는 영향)

  • Eun, Young-Joon;Song, Yun-Kyung;Lim, Hyung-Ho;Kwon, Ki-Rok;Rhim, Tae-Jin
    • Journal of Pharmacopuncture
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    • v.10 no.3
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    • pp.53-62
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    • 2007
  • Objective The objective of this study was to investigate the antioxidative effects of Puerariae Radix extract. Method Total antioxidant capacity (TAC), Total antioxidant response (TAR), Total phenolic content, Reactive oxygen species (ROS), 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activities, lipid peroxidation were examined. Result Total antioxidant status was examined by total antioxidant capacity(TAC) and total antioxidant response(TAR) against potent free radical reactions. TAC and TAR of Puerariae Radix extract at the concentration of 5 mg/ml were 2.02 and 1.50 mM Trolox equivalents, respectively. Total phenolic content of Puerariae Radix extract at the concentration of 5 mg/ml was 2.29 mM gallic acid equivalent. Concentration of Puerariae Radix extract at which DPPH radical scavenging activity was inhibited by 50% was 5.91 mg/ml as compared to 100% by pyrogallol solution as a reference. The inhibitory effect of the extract on lipid peroxidation was examined using rat liver mitochondria induced by FeSO4/ascorbic acid. Puerariae Radix extract at the concentration of 1 mg/ml slightly but significantly decreased TBARS concentration. The extract further prevented lipid peroxidation in a dose-dependent manner. The effect of Puerariae Radix extract on reactive oxygen species (ROS) generation was examined using cell-free system induced by hydrogen peroxide/FeSO4. Addition of 1 mg/ml of Puerariae Radix extract significantly reduced dichloroflurescein (DCF) fluorescence. The extract caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that the extract significantly prevented ROS generation in vitro. Thus antioxidant effects of Puerariae Radix extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation. Conclusion As a result, Puerariae Radix seems to have antioxitative effect and antioxidant compount.

A Study on Industrial Media for Production of Lactic acid in Batch and Continuous Fermentations (회분식 및 연속배양에 있어서 고농도 젖산의 생산을 위한 공업용 배지연구)

  • 김양훈;이기범;문승현
    • KSBB Journal
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    • v.14 no.2
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    • pp.181-187
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    • 1999
  • We have investigated industrial media for lactic acid fermentation to reduce the cost of nitrogen sources. Corn steep liquor (CSL) was successfully used at 5% (v/v) in batch fermentations. Use of soluble CSL improved the productivity about 20% with an advantage of clearer fermentation broth. Yeast extract-complemented CSL improved the productivity about 20% with an advantage of clearer fermentation broth. Yeast extract-complemented CSL media further increased the increased the productivity. It was found that 3.1 g/L yeast extract and 5% CSL could be an effective substitute for 15 g/L yeast extract in 10% glucose medium. Brewing yeast was also used as a sole nitrogen source equivalent to 5% CSL. A continuous culture coupled with cell-recycle by microfiltration at the dilution rate of 0.05-0.065 h-1 led to the highest lactic acid productivity. Lactic acid was recovered by electrodialysis from the cell free broth. Depleted cell free broth supplemented with 5-10 g/L of yeast extract performed reasonably in batch and continuous cultures. Reuse of the fermentation broth may reduce the cost of raw materials as well as minimize the fermentation wastes.

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Antioxidative Properties of Sachil-Tang Extract

  • Yi, Hyo-Seung;Moon, Jin-Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.4
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    • pp.872-882
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    • 2009
  • Sachil-Tang (SCT) has been traditionally used as a prescription of spasm of the esophagus by stress, pectoralgia and oppressive feeling of the chest in Oriental medicine. This study was carried out to investigate the antioxidant activities of the ethanol extract of SCT and its inhibitory effect on intracellular oxidation and vascular cell adhesion molecule-1 expression in human umbilical vein endothelial cells (HUVECs) using various methods. The SCT extract showed a strong inhibitory effect on free radical generating model systems, including DPPH radical, superoxide anions, hydroxyl radical, peroxynirite and nitric oxide. Besides, the SCT extract exhibited a strong inhibitory effect on lipid peroxidation in rat liver homogenate induced by $FeCl_2$-ascorbic acid, and protected plasmid DNA against the strand breakage in a Fenton's reaction system. The SCT extract also inhibited copper-mediated oxidation of human low-density lipoprotein (LDL), and repressed relative electrophoretic mobility of LDL. Furthermore, the SCT extract protected intracellular oxidation induced by various free radical generators and inhibited expression of vascular cell adhesion molecule-1 (VCAM-1) in HUVECs. These results suggest that SCT can be an effective natural antioxidant and a possible medicine of atherosclerosis.

Antioxidative Constituents from Paeonia lactiflora

  • Lee, Seung-Chul;Kwon, Yong-Soo;Son, Kyung-Hun;Kim, Hyun-Pyo;Heo, Moon-Young
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.775-783
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    • 2005
  • The ethanol extract of the peony root (Paeonia Lactiflora Pall, Paeoniaceae) as well as its major active components including gallic acid and methyl gallate were evaluated for their protective effects against free radical generation and lipid peroxidation. In addition, the protective effects against hydrogen peroxide-induced oxidative DNA damage in a mammalian cell line were examined. The ethanol extracts of the peony root (PREs) and its active constituents, gallic acid and methyl gallate, exhibited a significant free radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and had an inhibitory effect on lipid peroxidation, as measured by the level of malondialdehyde (MDA) formation. The PREs did not have any pro-oxidant effect. They strongly inhibited the hydrogen peroxide-induced DNA damage from NIH/3T3 fibroblasts, as assessed by single cell gel electrophoresis. Furthermore, the oral administration of 50% PRE (50% ethanol extract of peony root), gallic acid and methyl gallate potently inhibited the formation of micronucleated reticulocytes (MNRET) in the mouse peripheral blood induced by a $KBrO_3$ treatment in vivo. Therefore, PREs containing gallic acid and methyl gallate may be a useful antigenotoxic antioxidant by scavenging free radicals, inhibiting lipid peroxidation and protecting against oxidative DNA damage without exhibiting any pro-oxidant effect.

The Effects of Bamboo Extract on Human Melanocytes and B16 Melanoma Cells in vitro

  • Cho, Joon-Hwan;Lee, Ki-Moo;Kim, Nam-Soo;Kang, Won-Hyoung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.25 no.4 s.34
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    • pp.133-143
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    • 1999
  • To identify inhibitory effect of Bamboo extract on melanogenesis, the effect was compared with arbutin, ascorbic acid, hydroquinone, and kojic acid on the melanin biosynthesis in B16 mouse melanoma cells and cultured human melanocytes. The cell viability of the agent was tested on cultured human melanocytes. We also examined its. free radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl(DPPH). Bamboo extract showed considerable effect against melanin production and did not reduce cell viability at the concentration tested. It also showed potent free radical scavenging activity.

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The Effects of Bamboo Extract on Human Melanocytes and B16 Melanoma Cells In vitro

  • Cho, Joon-Hwan;Lee, Kim-Moo;Kim, Nam-Soo;Kang, Won-Hyoung
    • Proceedings of the SCSK Conference
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    • 1999.10a
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    • pp.133-143
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    • 1999
  • To identify inhibitory effect of Bamboo extract on melanogenesis, the effect was compared with arbutin. ascorbic acid, hydroquinone, and kojic acid on the melanin biosynthesis in Bl6 mouse melailoma cells and cultured hunlan melanocytes. The cell viability of the agent was tested on cultured human melanocytes. We also examined. its free radical scavenging activity on 1,1-dipheny1-2-picrylhydrazyl (DPPH). Bamboo extract showed considerable effect against melanin production and did not reduce cell viability at the concentration tested. It also showed potent free radical scavenging activity.

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Glutathione Sulphydryl Contents and Antioxidant Activities of Lactobacillus spp. and Bacillus coagulans (Lactobacillus spp.와 Bacillus coagulans의 Glutathione Sulphydryl 함유율과 황산화 활성)

  • Byun, Jeong-Yeol;Yoon, Yeong-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.22 no.2
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    • pp.91-97
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    • 2004
  • The antioxidative ability on the basis of reduced glutathione sulphydryl(GSH) level, the inhibition activities of linoleic acid peroxidation of cell free extract of Lactobacillus spp. and Bacillus spp. have been determined; Lactobacillus casei CU4114 contained the highest level of GSH among the probiotic strains with 25.15 ${\mu}$mole/g. Significantly high level of GSH occured in the intracellular cell free extract of Lactobacillus rhamnosus CU4201, Lactobacillus plantarum CU4203. The antioxidant activity and inhibition of linoleic acid peroxidation of cell free extract of Lactobacillus spp. and Bacillus spp. by thiobarbituric acid(TBA) assay have been shown to be significantly differed depending on the strains(P>0.01). Intracellular cell free extracts of L. acidophilus CU4111, L. casei CU4114, and strains of Bacillus coacillus revealed a significantly intensive inhibitory activity in the linoleic acid peroxidation reactions. Spearmans' rank correlation between inhibitory activity on linoleic acid peroxidation and cellular GSH levels of Lactobacillus spp. was analysed and the correlation quotient was 0.65 which means a significant positive correlation.

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Microbiological Characteristics and Physiological Functionality of New Records of Yeasts from Wild Flowers in Yokjido, Korea

  • Hyun, Se-Hee;Lee, Jong-Soo
    • Mycobiology
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    • v.42 no.2
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    • pp.198-202
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    • 2014
  • Two new yeast records, Cryptococcus adeliensis YJ19-2 and Cryptococcus uzbekistanensis YJ10-4 were screened from 60 yeasts strains that were isolated and identified from wild flowers in Yokjido, Gyeongsangnam-do, Korea. The morphological and cultural characteristics of the newly recorded yeasts and the physiological functionalities of the supernatants and cell-free extracts obtained from their cultures were investigated. The two newly recorded yeasts did not form ascospores and pseudomycelia. They also grew well in yeast extract-peptone-dextrose broth. C. uzbekistanensis YJ10-4 grew in a vitamin-free medium and was also tolerant to sugar and salt. Antihypertensive angiotensin I-converting enzyme inhibitory activity of the supernatant from C. adeliensis YJ19-2 was high (71.8%) and its cell-free extract also showed very high (81.2%) antidiabetic $\acute{a}$-glucosidase inhibitory activity.