• Journal of Acupuncture Research
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• v.19 no.2
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• pp.51-64
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• 2002

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.3
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• pp.287-294
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• 2015

In this study, we investigated the antioxidative effects of brown seaweed Ecklonia cava extract and its subfractions. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of E. cava. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of ethyl acetate fraction ($FSC_{50}=6.98{\mu}g/mL$) and aglycone fraction ($7.03{\mu}g/mL$) are similar to that of (+)-${\alpha}$-tocopherol ($8.98{\mu}g/mL$) which is a reference control. Reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) of the aglycone fraction ($OSC_{50}=14.48{\mu}g/mL$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was the strongest among all extract and fractions. However, all samples showed lower antioxidant activities than that of L-ascorbic acid ($6.88{\mu}g/mL$) known as a powerful antioxidant. The protective effect of 50% ethanol extract on the $^1O_2$-induced cellular damage of human erythrocytes was dependent on the concentration from 5 to $50{\mu}g/mL$. Both ethyl acetate fraction and aglycone fraction showed strong cellular protective activities at $10{\mu}g/mL$, where the cellular protective effects (${\tau}_{50}$) of each fraction were recorded 442.0 min and 539.9 min, respectively. Three kinds of extract/fractions of E. cava showed much greater cellular protective activities at $10{\mu}g/mL$ than that of liposoluble antioxidant (+)-${\alpha}$-tocopherol (40.6 min) which is a reference control. These results suggest E. cava extracts and its fractions can be applied as an antioxidant ingredient in a field of cosmetics.

• Korean Journal of Plant Resources
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• v.26 no.4
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• pp.441-449
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• 2013

This study was conducted to select some useful plants as functional material candidates. A total of 38 plants were preliminarily screened for the anti-inflammatory and antioxidant activities. The preliminarily selected 8 plants were further investigated to verify the in vitro inhibitory effect on inflammation and oxidative stress. Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) inhibited the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Eupatorium japonicum (leaf) suppressed the expression of cyclooxygenase-2 (COX-2), whereas Boehmeria platanifolia (root) and Prunus yedoensis (branch) inhibited the transcription of nuclear factor-kappa B (NF-${\kappa}B$). Treatment with the extracts ($2.5{\sim}20{\mu}g/ml$) of Abutilon theophrasti (leaf, flower/seed) and Hemistepta lyrata (stem) did not show toxicity on RAW 264.7 cell proliferation, but treatment with $2.5{\mu}g/ml$ of Boehmeria platanifolia (root) exhibited cell toxicity. Carpinus coreana (branch) and Prunus yedoensis (branch) showed potent scavenging activities on peroxynitrite. Akebia quinata (flower), Carpinus coreana (branch), and Prunus yedoensis (branch) effectively inhibited reactive oxygen species (ROS). Abutilon theophrasti (leaf), Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) exhibited strong inhibitory capacity with regard to nitric oxide (NO) production. The results suggested that Abutilon theophrasti (leaf) has in vitro anti-inflammatory and antioxidant activities, and that is a useful functional material candidate.

• Food Science and Preservation
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• v.24 no.3
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• pp.446-454
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• 2017

Garlic (Allium sativum L.) and traditional herb has several functional properties and strong biological activities, making it useful as a functional food material. We investigated the antioxidant and anti-inflammatory activity of mixed compounds from red garlic and supplementary materials, including ginger (Zingiber officinale Roscoe), doraji (Platycodon grandiflorum), quince (Chaenomeles sinensis), citrus peel (Citri Pericarpium), and mint (Mentha arvensis). The extracts were prepared with water (W) and ethanol (E) at $70^{\circ}C$ (W-70, E-70) and $95^{\circ}C$ (W-95, E-95) for 3 h. The total content of phenolic compounds was the highest in E-70 (608.60 mg/100 g). Alliin, one of the active ingredients in red garlic, was contained at 1.18-1.29 mg/g and 0.81-0.85 mg/g in water and ethanol extract, respectively. Another active ingredient of red garlic, S-allyl-cysteine (SAC) had higher content in the water extract than in the ethanol extracts. DPPH radical scavenging activity was higher in E-70 (15.96-73.65%) at $313-5,000{\mu}g/mL$. ABTS radical scavenging activity was also higher in E-70 (5.71-77.19%) than in the others. The ROS production rate showed the same tendency as the NO production, with more efficacy in E-95. The expression level of iNOS and $IL-1{\beta}$ was decreased in the E-95 significantly at the concentration of $1,000{\mu}g/mL$ compared to the lipopolysaccharide (LPS) treated group. Based on the above results, the antioxidative and anti-inflammatory activities of the extracts of red garlic and supplementary materials were expressed by different useful substances. The contents of these useful substances were different according to the extraction solvent and temperature.

• Korean Journal of Food Science and Technology
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• v.53 no.4
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• pp.423-433
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• 2021

This study was performed to examine the neuroprotective effect of the ethyl acetate fraction from Eucommia ulmoides oliver leaf (EFEL) on PM_2.5-induced cytotoxicity. EFEL had higher total phenolic and flavonoid contents than the other fractions. In ABTS and DPPH radical scavenging activities, the IC₅₀ of EFEL was measured as 212.80 and 359.13 ㎍/mL, respectively. To investigate the neuroprotective effect of EFEL, MTT and DCF-DA assays were performed on HT22, MC-IXC, and BV-2 cells. EFEL effectively decreased PM_2.5-induced intercellular reactive oxygen species (ROS) content and inhibited PM_2.5-induced cell death. In the results of protein expression related to cellular cytotoxicity on microglial cells (BV-2), EFEL had an improvement effect on cell apoptosis and inflammatory pathways. Rutin and chlorogenic acid were identified as the main physiological compounds. Moreover, it was expected that EFEL, including rutin and chlorogenic acid, could be functional food substances with neuroprotective effects against PM_2.5-induced oxidative stress.

• Applied Chemistry for Engineering
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• v.29 no.6
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• pp.772-781
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• 2018

In this study, we investigated antioxidant, antimicrobial and cytoprotective effects of 50% ethanol extract and ethyl acetate fraction from rhizomes of Belamcanda chinensis (L.) DC. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activities ($FSC_{50}$) of the 50% ethanol extract and ethyl acetate fraction were 621.5 and $253.0{\mu}g/mL$, respectively. Total antioxidant capacities ($OSC_{50}$) of the extract and fraction were 13.6 and $3.0{\mu}g/mL$, respectively. Minimum inhibitory concentrations (MIC) of the ethyl acetate fraction for Staphylococcus aureus and Candida albicans were 156, $1,250{\mu}g/mL$, respectively, indicating similar or higher levels of those of using methyl paraben. Cytoprotective effects of the 50% ethanol extract against $^1O_2$-induced cellular damage (${\tau}_{50}$) showed in a dose dependent manner at 4 to $64{\mu}g/mL$. ${\tau}_{50}$ of the 50% ethanol extract, ethyl acetate fraction and (+)-${\alpha}$-tocopherol at $16{\mu}g/mL$ were 36.4, 45.0 and 45.8 min respectively, and the ethyl acetate fraction showed cytoprotective effects similar to (+)-${\alpha}$-tocopherol. In ultraviolet B radiation-induced HaCaT cell damage, the ethyl acetate fraction decreased intracellular reactive oxygen species (ROS) up to 45.9% at $8{\mu}g/mL$. Also in $H_2O_2$-induced HaCaT cell damage, the ethyl acetate fraction significantly increased the cell viability at $0.5{\sim}8.0{\mu}g/mL$. As a result of chemical analyses of the ethyl acetate fraction, the presence of flavonoids and polyphenol such as irisflorentin, irigenin, tectorigenin, resveratrol, iridin and tectoridin were identified. In conclusion, the extract/fraction from rhizomes of B. chinensis can be applied as a natural antioxidant and antimicrobial material to cosmetics.

• Journal of Life Science
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• v.29 no.5
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• pp.596-606
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• 2019

The study investigated the physiochemical properties and the antioxidant and anti-inflammatory activities of the sea tangle (Saccharina japonica) in a water extract before (STWE) and after (STFL) fermentation with Lactobacillus brevis. The pH values of STWE and STFL were 6.18 and 4.16, and the sugar contents were $8.50^{\circ}Brix$ and $7.40^{\circ}Brix$, respectively. The main free amino acids of STWE and STFL were glutamic acid, aspartic acid, and alanine, and the ${\gamma}$-amino butyric acid (GABA) content was increased by fermentation. The total polyphenol contents of STWE and STFL were 498.29 and 615.77 mg gallic acid equivalent (GAE)/ml, respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of STWE and STFL were markedly increased in a dose-dependent manner, and revealed about 89.89% and 96.94% activities, respectively, at 10% concentration (p<0.05). The superoxide dismutase (SOD) activities of STWE and STFL were also markedly increased in a dose-dependent manner, and the activity of STFL was significantly increased when compared with STWE (p<0.05). The anti-inflammatory activity was examined in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. STWE and STFL decreased the production of reactive oxygen species (ROS), which had levels of about 189.90% and 174.69% at 1% concentration, respectively (p<0.05). The contents of pro-inflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-{\alpha}$) and interleukin-6 (IL-6), were decreased more by addition of STFL than by addition of STWE. The STWE and STFL showed high antioxidant and anti-inflammatory activity, and these activities were increased by fermentation. Therefore, sea tangle extracts can be used as functional food materials.

• Biomolecules & Therapeutics
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• v.21 no.2
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• pp.146-152
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• 2013

This study examined the total polyphenol content of eight wild edible plants from Ethiopia and their effect on NO production in Raw264.7 cells. Owing to its relatively high polyphenol concentration and inhibition of NO production, the methanol extract of Adansonia digitata L. leaf (MEAD) was subjected to detailed evaluation of its antioxidant and anti-inflammatory effects. Antioxidant effects were assessed by measuring free-radical-scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and oxygen-radical-absorbance capacity (ORAC) assays, while anti-inflammatory effects were assessed by measuring inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In the ORAC assay, MEAD was 10.2 times more potent than vitamin C at eliminating peroxyl radicals. In DPPH assay, MEAD also showed a strong ROS scavenging effect. MEAD significantly inhibited iNOS activity ($IC_{50}=28.6{\mu}g/ml$) of LPS-stimulated Raw264.7 cells. We also investigated the relationship between iNOS expression and nuclear factor kappa B (NF-${\kappa}B$) activation. MEAD inhibited $I{\kappa}B{\alpha}$ degradation and NF-${\kappa}B$ translocation from the cytosol to the nucleus in LPS-induced RAW264.7 cells without significant cytotoxic effects, as confirmed by MTT assay. These results suggest that MEAD inhibits anti-inflammatory iNOS expression, which might be related to the elimination of peroxyl radicals and thus the inhibition of $I{\kappa}B{\alpha}$-mediated NF-${\kappa}B$ signal transduction.

• Korean Journal of Medicinal Crop Science
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• v.17 no.4
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• pp.243-250
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• 2009

Fifty percent ethanol extract of Lythrum salicaria Linne root (LSR) was tested in vitro on antioxidant activity, and furthermore was investigated on antioxidative and fibrosis protecting activities in $CCL_4$-induced liver fibrosis rat model. Ratio of hepatic GSH/GSSG (reduced glutathione/oxidized glutathione) as bio-parameter of antioxidant level in $CCL_4$ plus LSR-treated rats for 6 weeks significantly increased from 2.8- to 5.7-fold than that of $CCL_4$-treated rats at p < 0.05. Thiobarbituric acid reactive substances (TBARS) contents in $CCL_4$ plus LSR-treated rats ranged from 1.57- to 2.19-fold of normal rats and were lower than those in $CCL_4$ plus silymarin-treated rats ($1.78{\sim}2.46$-fold of normal rats) (p < 0.05). Amounts of hydroxyproline of liver tissue showing the content of total collagen, a parameter of fibrosis, in $CCL_4$ plus LSR-administrated rat livers were $4.9{\sim}8.8{\mu}g$/mg ($-47{\sim}-71%$, compared with that in $CCL_4$-treated rat livers ($16.6{\mu}g$/mg tissue), which were significantly lower than those in $CCL_4$ plus silymarin-administrated rats being $8.4{\sim}11.7{\mu}g$/mg ($-30{\sim}-50%$). This collagen reducing effect of liver tissue in $CCL_4$ plus LSR-treated rats was supported by histological observation using microscopy assay. From the results, we conclude that the root of L. salicaria have efficient antioxidant potential and effective antifibrotic activities.