• Journal of Applied Biological Chemistry
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• v.54 no.3
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• pp.171-177
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• 2011

This study was carried out to search for natural anti-oxidants and anti-inflammatory compounds from 5 medicinal plants (Carthamus tinctorius seed, Cyperus rotundus, Schizonepeta tenuifolia, Polygonatum odoratum var. pluriflorum, and Paeonia lactiflora). These plants were extracted with 70% ethanol. In order to measure total antioxidant activity of flavonoids, polyphenol content was measured. Radical scavenging activities of extracts were examined using a-a-Diphenyl-${\beta}$-picrylhydrazyl ($DPPH{\cdot}$), 2,2-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid ($ABTS{\cdot}$), ferric reducing antioxidant power (FRAP) and superoxide anion radical assays. C. tinctorius seed extracts showed the highest polyphenol and flavonoid contents as well as strong $DPPH{\cdot}$, $ABTS{\cdot}$, FRAP, and superoxide anion radical scavenging activity. Also, C. tinctorius seed extracts showed the highest nitric oxide (NO) production inhibitory effect. Theses results indicate that the C. tinctorius seed extracts can be used as a functional material due to their effective anti-oxidative and antiinflammatory activities.

• The Korea Journal of Herbology
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• v.33 no.2
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• pp.19-28
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• 2018

Objectives : Epimedii Herba has been frequently used in Korean Traditional Medicine to treat impotence, spermatorrhoea, exophthalmos, and forgetfulness. Present study investigated anti-inflammatory effects of Epimedii Herba water extract (EWE) and attempted to elucidate molecular mechanisms involved. Methods : To explore anti-inflammatory effects of EWE, RAW 264.7 cells, a murine macrophage cell line, were pretreated with $10-100{\mu}g/m{\ell}$ of EWE, and then subsequently exposed to $1{\mu}g/m{\ell}$ of lipopolysaccharide (LPS). Levels of nitric oxide (NO), interleukin-6, $interleukin-1{\beta}$, and tumor necrosis $factor-{\alpha}$ were monitored in the medium. Expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by immunoblot and real-time PCR analyses. Signaling pathways related with nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinases were monitored to elucidate molecular mechanisms involved. Finally, the role of three flavonoid compounds in EWE on LPS-mediated NO production were investigated. Results : In conditioned medium, pretreatment of EWE ($100{\mu}g/m{\ell}$) significantly inhibited LPS-stimulated NO and pro-inflammatory cytokine production. In addition, EWE attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by LPS. EWE prevented the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}$, nuclear translocation of $NF-{\kappa}B$, and DNA binding of $NF-{\kappa}B$, while EWE did not change the phosphorylation of mitogen-activated protein kinases by LPS. Moreover, icariin, icaritin, and quercetin partly, but significantly, inhibited the LPS-stimulated NO production. Conclusions : These results suggest that EWE has an ability to prevent inflammation in macrophages through inhibition of $NF-{\kappa}B$ signaling pathway.

• Journal of Life Science
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• v.33 no.1
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• pp.73-81
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• 2023

In the present study, we prepared hot water extracts and the subsequent organic solvent fractions of methanol extracts of Houttuynia cordata (HC) and Lespedeza cuneata (LC), and investigated their anti-inflammatory effects on lipopolysaccharide-stimulated RAW264.7 cells. Among the treated samples, hexane, chloroform, and ethyl-acetate fractions of HC and LC inhibited nitric oxide (NO) production in a dose-dependent manner, and decreased inducible nitric oxide synthase (iNOS) protein expression. And, we analyzed the flavonoid contents of the ethyl-acetate fraction of HC and LC, and chose apigenin for the further experiments because apigenin was one of flavonoids commonly found in HC and LC. Apigenin dramatically inhibited NO production in a dose-dependent manner without affecting cell viability and decreased iNOS and cyclooxygenase-2 (COX-2) expression. In addition, apigenin suppressed the phosphorylation of p38 and Jun N-terminal kinase (JNK) indicating that apigenin exerts anti-inflammatory activity via the mitogen-activated protein kinase (MAPK) signaling pathway. Subsequently, we conducted RNA-sequencing analysis to detect differentially expressed genes upon apigenin treatment. Among the down-regulated genes, four cytokine genes (interleukin (IL)-1α, IL-1β, IL-6, and colony stimulating factor 2 (CSF2)) were selected for the further analysis, and the reduction of their expression by apigenin was confirmed with quantitative real-time polymerase chain reaction. Overall, our results suggest that Houttuynia cordata and Lespedeza cuneata have the anti-inflammatory effects and apigenin can be the one of key molecules responsible for their anti-inflammatory activities.

• Journal of the Korean Applied Science and Technology
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• v.36 no.4
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• pp.1485-1495
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• 2019

The purpose of this study was to evaluate the antioxidant and anti-inflammatory effects of extracts of callus cultures, pericarp, flesh, fruit of Trapa Japonica. The toxicity of extracts from Trapa Japonica pericarp investigated using the RAW 264.7 cell showed 45.8 ± 1.5% of cell survival rate. And the results of investigation using HaCaT cells showed a 51.1 ± 1.0% cell viability at 100 ㎍/㎖ in the pericarp extract and lower cell viability at higher concentrations. The total content of polyphenol pericarp extract was 213.20 ± 15.78 mg/g, while the total content of flavonoid was 29.30 ± 3.24mg. And the total content of polyphenol callus cultures extract was 205.20 ± 18.97 mg/g, while the total content of flavonoid was 237.4 ± 7.43 mg. With a concentration level of 0.005 ~ 1000 ㎍/㎖ extract of Trapa Japonica pericarp the range of removal of 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radicals was 67.53 ± 1.5 % ~ 75.75 ± 0.5 % respectively and the range of removal of extract of Trapa Japonica callus cultures extract was also 3.1 ± 0.1 % ~ 77.32 ± 0.5 % respectively. As a result of measuring the Nitric Oxide(NO) generation amount of all Trapa Japonica extract 6.25, 12.5, 25, 50 ㎍/㎖ concentration exhibited significant (p < 0.05, p < 0.01) decreases.

• Journal of Plant Biotechnology
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• v.47 no.3
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• pp.254-259
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• 2020

We studied the antioxidative and immunoregulatory properties of 70% ethanol extracts from the leaves, branches, and fruit of 13 species of the Rhamnaceae collected during the bearing season (Berchemia berchemiifolia, Berchemia floribunda, Hovenia dulcis, Paliurus ramosissimus, Rhamnella franguloides, Rhamnus crenata, Rhamnus davurica, Rhamnus koraiensis, Rhamnus parvifolia, Rhamnus ussuriensis, Rhamnus yoshinoi, Sageretia thea, and Ziziphus ujube). Total phenolic and flavonoid content values were determined using calibration curves of gallic acid and rutin, respectively. ABTS and DPPH radical scavenging assays were performed to determine antioxidative potential. Anti-inflammatory effects were studied using the nitric oxide (NO) inhibition assay in LPS-induced RAW264.7 cells. Extracts of Berchemia berchemiifolia, Sageretia thea, and Ziziphus jujuba had the highest total phenol and flavonoid content values. Extracts of Berchemia berchemiifolia, Paliurus ramosissimus, Rhamnella franguloides, and Sageretia thea had the highest ABTS radical scavenging activities. Berchemia berchemiifolia and Sageretia thea had the highest DPPH radical scavenging activities. Furthermore, extracts of Berchemia berchemiifolia, Berchemia floribunda, Paliurus ramosissimus, Sageretia thea, and Ziziphus jujuba showed strong anti-inflammatory activity through inhibition of NO production. The results of this study suggest that Rhamnaceae extracts can be used as natural antioxidants and immunomodulators.

• Journal of Life Science
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• v.31 no.2
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• pp.158-163
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• 2021

In the present study, we prepared hot water extracts of green apple (GAHW) and unripe apple (UAHW), and ethanol extract of green apple (GAE), and investigated their anti-inflammatory activities in LPS-activated RAW264.7 cells. All extracts dramatically suppressed nitric oxide (NO) production in a dose-dependent manner in LPS-stimulated RAW264.7 cells without affecting cell viability. In addition, all extracts decreased the expression of iNOS, whereas UAHW only reduced the expression of COX-2. All extracts suppressed the phosphorylation of MAPKs (p38, ERK, and JNK) indicating all extracts show their anti-inflammatory activities via regulating MAPK pathway. Furthermore, all extracts reduced the production of reactive oxygen species in a dose-dependent manner and they increased the expression of heme oxygenase-I (HO-I) whereas UAHW could not. We also investigated whether apple flavonoids phloretin and phloridzin can have their anti-inflammatory activities in same in vitro model. Phloretin dramatically decreased NO production in a dose dependent manner without affecting cell viability, whereas phloridzin have no effects. Phloretin also reduced the expression of iNOS as well as COX-2, whereas phloridzin could not. Overall, these results suggest that apple extracts have their anti-inflammatory activities via regulating MAPKs and HO-1 pathways, and apple flavonoid phloretin can be one of phytochemicals responsible for anti-inflammatory effect of apple.

• Journal of Life Science
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• v.29 no.12
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• pp.1371-1377
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• 2019

Oxidative stress induced by the over-production of reactive oxygen species (ROS) in the brain is the most common cause of neurodegenerative diseases such as Alzheimer's. In the present study, we investigated the protective effects of flavonoids and their glycosides, namely kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-β-D-glucoside, against H₂O₂-induced oxidative stress in the C6 glial cells. The H₂O₂-treated glial cells exhibited decreased cell viability and increased ROS production when compared with normal cells. However, cells treated with each of the four flavonoids/glycosides demonstrated significantly increased viability and suppressed ROS production when compared with the H₂O₂-treated control group. These results indicate that flavonoids/glycosides attenuate oxidative stress induced by H₂O₂ in C6 glial cells. To confirm the protective molecular mechanisms, we measured pro-inflammatory factors such as inducible nitric oxide synthase, cyclooxygenase-2, and interleukin-1β. H₂O₂ treatment was seen to elevate these factors and decrease IκB-α in the C6 glial cells, while the flavonoids/glycosides induced a down-regulation of the pro-inflammatory factors and increased IκB-α, indicating a neuroprotective effects through attenuation of the inflammation. In particular, quercetin and its glycoside showed a higher neuroprotective effect than the kaempferol treatments. These results suggest that these flavonoids and their glycosides could be promising therapeutic agents for neurodegenerative diseases via the attenuation of oxidative stress.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.327-332
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• 2019

Alzheimer's disease (AD) is a common neurodegenerative disease. Oxidative stress by amyloid beta peptide (Aβ) of neuronal cell is the most cause of AD. In the present study, protective effects of several flavonoids such as kaempferol (K), kaempferol-3-O-glucoside (KG), quercetin (Q) and quercetin-3-β-ᴅ-glucoside (QG) from Aβ_25-35 were investigated using C6 glial cell. Treatment of Aβ_25-35 to C6 glial cell showed decrease of cell viability, while treatment of flavonoids such as Q and QG increased cell viability. In addition, treatment of flavonoids declined reactive oxygen species (ROS) production compared with Aβ_25-35-induced control. The ROS production was increased by treatment of Aβ_25-35 to 133.39%, while KG and QG at concentration of 1 μM decreased ROS production to 107.44 and 113.10%, respectively. To study mechanisms of protective effect of these flavonoids against Aβ_25-35, the protein expression related to inflammation under Aβ_25-35-induced C6 glial cell was investigated. The results showed that C6 glial cell under Aβ_25-35-induced oxidative stress up-regulated inflammation-related protein expressions. However, treatment of flavonoids led to reduction of protein expression such as inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1β. Especially, treatment of KG and QG decreased more effectively inflammation-related protein expression than its aglycones, K and Q. Therefore, the present results indicated that K, Q and its glycosides attenuated Aβ_25-35-induced neuronal oxidative stress and inflammation.

• Food Science and Preservation
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• v.25 no.1
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• pp.90-97
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• 2018

Peach seeds contain a large amount of phenolic components and exhibit excellent physiological effects in various diseases. We examined the antioxidant effects of stone and seed of three peach cultivars (Miwhang, MH; Kanoiwa hakuto, KH; and Cheonhong, CH) by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, ferric reducing activity of plasma (FRAP) assay, and cupric ion reducing antioxidant capacity (CUPRAC) reduction. The results showed that the stone extracts of CH had higher levels of total phenols and flavonoids than those of the other cultivars do, and the stone extracts of KH and CH have the potential to reduce DPPH, FRAP, and CUPRAC activities. In addition, we found that KH, MH, and CH stone extracts decreased nitric oxide generation in RAW 264.7 and BV2 cells. The total phenol and flavonoid contents had no significant correlation with anti-oxidant activities. On the other hand, the anti-inflammatory activity had a low linear correlation with anti-oxidant activities and total phenol and flavonoid contents. The present results suggest that the correlation between antioxidant and anti-inflammatory effects of stone and seed, and the appropriate combination of the stone and seed extracts could be used as an anti-inflammatory treatment and prevention material, respectively.

• Advances in Traditional Medicine
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• v.8 no.3
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• pp.266-278
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• 2008

Erythrina stricta, a deciduous tree widely used traditionally in indigenous system of medicine for various ailments such as rheumatism, fever, leprosy, epilepsy etc. The leaves of Erythrina stricta was extracted with ethanol (70%) and used for the evaluation of various in vitro antioxidant assays which includes H - donor activity, nitric oxide scavenging, superoxide anion scavenging, reducing ability, hydroxyl radical, hydrogen peroxide scavenging, total phenolic content, total flavonoid content, total antioxidant activity by thiocyanate and phosphomolybdenum method, metal chelating, $\beta$-carotene bleaching, total peroxy radical assays. The pro-oxidant activity was measured using bleomycin-dependent DNA damage. Ex vivo models like lipid peroxidation and erythrocyte haemolysis were also used to study the antioxidant property of the extract. The various antioxidant activities were compared with suitable standard antioxidants such as ascorbic acid, butylated hydroxyl toluene, $\alpha$-tocopherol, curcumin, quercetin and Trolox. The generation of free radicals viz. $O_2^{{\cdot}-}$, $OH^{\cdot}$, $H_2O_2$, $NO^{\cdot}$ and peroxyl radicals were effectively scavenged by the ethanolic extract of Erythrina stricta. In all the methods, the extract offered strong antioxidant activity in a concentration dependent manner. The total phenolic content, flavonoid content and total antioxidant activity in Erythrina stricta were determined as microgram (g) pyrocatechol, quercetin and $\alpha$-tocopherol equivalent/mg respectively. The extract did not exhibit any prooxidant activity when compared with ascorbic acid. The results obtained in the present study clearly indicates that Erythrina stricta scavenges free radicals and reduces lipid peroxidation, ameliorating the damage imposed by oxidative stress in different disease conditions and serve as a potential source of natural antioxidant.