• Journal of Life Science
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• v.29 no.11
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• pp.1218-1226
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• 2019

The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

• Food Science and Preservation
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• v.25 no.1
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• pp.107-116
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• 2018

The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and prostaglandin $E_2$ ($PGE_2$) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and $ABTS^+$ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were $217.04{\pm}2.98$, $156.72{\pm}3.90$, and $182.88{\pm}3.02mg\;TAE/g$, respectively, while the electron donating abilities at $1,000{\mu}g/mL$ of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The $ABTS^+$ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at $50{\mu}g/mL$ of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

• Journal of Life Science
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• v.29 no.3
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• pp.325-331
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• 2019

Lettuce (Lactuca sativa L.) is one of the most popular green leafy vegetables, and it contains various beneficial components including polyphenolic compounds and has been known to possess various biological functions such as anti-microbial, anti-oxidative, and anti-inflammatory activities. In the present study, we prepared ethanol extract of dried lettuce (DLE) and investigated its anti-inflammatory activity. To evaluate the anti-inflammatory activity of DLE, nitric oxide (NO) production was measured in LPS-activated mouse macrophage RAW 264.7 cells. DLE significantly suppressed NO production in these cells without affecting cell viabilities while resveratrol was used as a positive control. DLE dramatically decreased the expression of pro-inflammatory genes such as iNOS and COX-2 at the mRNA and protein levels and reduced the expression of several cytokines including $IL-1{\alpha}$, $IL-1{\beta}$, IL-1F6, $TNF-{\alpha}$, CSF2 and CXCL10. In addition, DLE suppressed phosphorylation of MAPKs and the nuclear translocation of $NF-{\kappa}B$ p65 indicating DLE shows its anti-inflammatory activity via regulating MAPKs pathway and $NF-{\kappa}B$ pathways. And also, DLE reduced the production of reactive oxygen species in a dose-dependent manner. DLE increased HO-1 protein expression, and also increased the nuclear translocation of Nrf2. Overall, our results suggest that lettuce down-regulate various pro-inflammatory genes and have its anti-inflammatory activity via regulating MAPKs, $NF-{\kappa}B$, and Nrf2/HO-1 pathways.

• Journal of Life Science
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• v.28 no.11
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• pp.1361-1368
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• 2018

Inflammation is the most common condition in the human body. Tissue damage triggers inflammation, together with vasodilation and increased blood flow at the inflamed site, resulting in edema. Inflammatory responses are also triggered by lipopolysaccharide (LPS), a Toll-like receptor Enterococcus faecalis, a gram-positive organism, has been reported to possess immunomodulatory and preventive activities; however, its use may present risks of sepsis and other systemic infections. Heat-killed Enterococcus faecalis (EF-2001) has been reported to induce antitumor activity, but its effects on inflammation are not known. In the present study, we investigated the effect of EF-2001 on LPS-induced macrophage inflammatory responses. EF-2001 treatment reduced nitric oxide (NO) production, indicating suppression of inflammatory reactions. EF-2001 showed no cytotoxicity in macrophages. Further investigation of the anti-inflammatory mechanism of EF-2001 indicated that EF-2001 reduced the LPS-induced expression of inducible nitric oxide synthase and cyclooxygenase-2. EF-2001 also reduced f the LPS induction of several inflammatory molecules involved in the nuclear factor-${\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinase pathways, including ERK, JNK, and p38 phosphorylation, in a concentration-dependent manner. Additionally, EF-2001 inhibited Akt phosphorylation and increased the expression of the inhibitory ${\kappa}B$ ($I{\kappa}B$) protein, an inhibitor of $NF-{\kappa}B$. EF-2001 also inhibited the nuclear translocation of p65. These results suggest that EF-2001 has anti-inflammatory properties and may be useful for treating inflammatory diseases.

• Korean Journal of Acupuncture
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• v.35 no.4
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• pp.166-173
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• 2018

Objectives : Woogakseungmatang is a prescription medication mainly used to treat facial paralysis in Korean medicine. The purpose of this study is to investigate the effects of Woogakseungmatang on anti-inflammation and anti-oxidation. Methods : Woogakseungmatang was extracted using hot water. Cytotoxicity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method; nitric oxide(NO) production and Prostaglandin $E_2$ ($PGE_2$) production in RAW cells treated with Woogakseungmatang were investigated; and the cytokine changes associated with inflammation were examined. The antioxidant capacity of Woogakseungmatang was measured using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Results : RAW cells treated with Woogakseungmatang showed 90% cell viability at a $100-{\mu}g/ml$ concentration. NO production was decreased by 15% at a $100-{\mu}g/ml$ concentration. $PGE_2$ production was decreased by 18% at a $100-{\mu}g/ml$ concentration. Interleukin $1{\beta}$ ($IL-1{\beta}$), interleukin 6(IL-6), and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) were significantly reduced at $100{\mu}g/ml$ compared with those in the control group. The DPPH free radical scavenging capability was more than 50% at $100{\mu}g/ml$. Conclusions : Woogakseungmatang showed only a slight anti - inflammatory effect at $100{\mu}g/ml$ and it was difficult to confirm the concentration-dependent anti-inflammatory effect. Therefore, this study means to confirm the potential anti-inflammatory effects of Woogakseungmatang. Based on this research, more systematic and diverse studies should be conducted.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.132-138
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• 2019

Among several marine-derived microorganisms isolated from the coast of Jeju Island that had antimicrobial activity against fish disease pathogens, Bacillus subtilis MD-02 was tested for its dietary effect on the innate immune response and disease resistance of olive flounder. Strain MD-02 was fed to the olive flounder at a concentration of $1.2{\times}10^4$, $1.2{\times}10^6$, or $1.2{\times}10^8CFU/100g$, respectively. Consequently, the hematocrit was higher in these three groups than that in the control group at 4 weeks, and the aspartate aminotransferase and alanine aminotransferase levels were decreased in the $1.2{\times}10^8$ and $1.2{\times}10^4CFU/100$ groups compared with the control group levels. The amylase activity and total protein were significantly increased in the $1.2{\times}10^4CFU/100g$ group at 3 weeks. The innate immune response, determined from the lysozyme and macrophage activities, was higher in the $1.2{\times}10^8CFU/100g$ group than in the control group. In addition, treatment of the olive flounders with Streptococcus parauberis at $1.2{\times}10^6CFU/ml$ confirmed the mortality rate, which was 100% in the control group and 40-60% in the groups fed B. subtilis MD-02, indicating that the fish had resistance to fish disease pathogens. Therefore, it was confirmed that when fed MD-02, olive flounder builds an innate immune response and acquires resistance to fish disease pathogens, indicating that B. subtilis MD-02 can be developed as a beneficial feed additive.

• Journal of Life Science
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• v.31 no.11
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• pp.987-994
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• 2021

Our previous studies have reported that antimicrobial peptides (AMPs) derived from the larvae of white-spotted flower chafer (Protaetia brevitarsis seulensis) exert anti-inflammatory and neuroprotective activities. This study explored the anti-inflammatory effects of protaetiamycine 9 (CVLKKAYFLTNLKLRG-NH₂), a novel AMP, derived from P. b. seulensis against lipopolysaccharide (LPS)-mediated inflammatory response in RAW264.7 macrophage cells. Protaetiamycine 9 (25, 50, 75, and 100 ㎍/ml) did not cause cytotoxic effects against RAW264.7 cells. The RAW264.7 cells were pre-treated with various concentrations of protaetiamycine 9 (25-100 ㎍/ml) for 1 hr and then exposed to LPS (100 ng/ml) for 24 hr. Protaetiamycine 9 treatments decreased the LPS-induced secretion of inflammatory mediators, such as nitric oxide (NO), in a dose-dependent manner. Protaetiamycine 9 (25-100 ㎍/ml) effectively downregulated the LPS-induced increase in mRNA and the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. Protaetiamycine 9 also suppressed the production and gene expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, compared to the presence of LPS alone. Furthermore, protaetiamycine 9 inhibited the degradation of inhibitory kappa B alpha (IκB-α) and the phosphorylation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. In conclusion, these results suggest that protaetiamycine 9 exhibits LPS-mediated inflammatory responses by blocking IκB-α degradation and MAPK phosphorylation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.4
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• pp.373-383
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• 2022

Inflammation caused by active oxygen and the resulting barrier damage have been consistently pointed out as the cause of wrinkle formation. In this study, effective index ingredient search and efficacy analysis were performed to verify the value of use as a functional cosmetic material related to antioxidant, anti-inflammatory and skin barrier improvement, and anti-aging for extracts of four types of Eleutherococcus divaricatus var. chiisanensis (ED), Eleutherococcus senticosus (EN), Eleutherococcus sessiliflorus (ES), and Eleutherococcus sieboldianus (EI) belonging to the Eleutherococcus genus. To identify the effective index composition, the content of the ingredients was measured by high-performance liquid chromatography. The content of eleutheroside E and chlorogenic acid was the highest in ED among the Eleutherococcus genus. As for anti-oxidant activity, DPPH radical scavenging activity was the highest in ED. In anti-inflammatory effects, ED extracts inhibited nitric oxide generation in inflammatory macrophage cells due to lipopolysaccharide by 40% at 100 ㎍/mL. In the case of IL-6 inhibition, which is known as a pro-inflammatory cytokine, ED showed 41% inhibition at 100 ㎍/mL. In addition, filaggrin and involucrin, which are skin barrier-related factors, were increased by 2.5 times and 1.6 times, respectively, in 100 ㎍/mL of ED extracts, and as for the collagenase, which is a wrinkle-related factor, ED extract showed 29% efficacy at 100 ㎍/mL. Thus, these result suggested that ED extract, among the four Eleutherococcus genus, can be used as a cosmetic ingredient for suppressing inflammation in the skin, reinforcing the skin barrier, and reducing wrinkles.

• The Korea Journal of Herbology
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• v.27 no.6
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• pp.115-122
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• 2012

Objectives : This study evaluated activities and ingredient contents concerning extracts according to extraction solvents of Insampaedok-san (IS, Renshen bai du-san). Methods : The herbal constituents of IS were extracted with water and 70% ethanol at $100^{\circ}C$ for 2 hr. Using the HPLC system, the six ingredient contents of different solvent extracts of IS were analyzed. The nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) production and proinflammatory cytokines were measured in RAW264.7 cells stimulated with lipopolysaccharide (LPS). The macrophage-derived chemokine (MDC/CCL22) and regulated on activation normal T-cell expression and secreted (RANTES/CCL5) production were measured in HaCaT and BEAS-2B cells stimulated tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$). The activities of glycerol-3-phosphate dehydrogenase (GPDH) and leptin level were measured in 3T3-L1 cells. Results : The calibration curves showed good linearity ($r^2$=1.0000) for different concentration ranges. The contents of liquiritin, naringin, hesperidin, neohesperirin and glycyrrizin in 70% ethanol extracts of IS were relatively higher than that of water extract, however the content of ferulic acid in 70% ethanol and water extract of IS were similar. The extraction solvents of water and 70% ethanol were evaluated inhibitory effect on the production of NO, $PGE_2$, TNF-${\alpha}$ and IL-6 in RAW 264.7 cells. Their extractions were inhibitory effect on production of MDC/CCL22 and RANTES/CCL5 in HaCaT cell and BEAS-2B cell, respectively. In addition, evaluated reduced on GPDH activity and leptin level in 3T3-L1 preadipocyte cell. Conclusions : Our results suggest that IS extracts were inhibitory effects of disease such as inflammation, allergies and obesity.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.421-427
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• 2022

In this study, Jeju Tatary buckwheat tea's chemical composition and physiological activities were compared according to the leaching temperature (60, 80, 100 ℃). As the leaching temperature is increased, the degree of browning is induced. However, there was no significant change in pH. The total polyphenol content was higher at 80 ℃ than at 60 ℃ leaching temperature, but significantly decreased at 100 ℃ leaching temperature (60 ℃: 17.06 mg GA/g, 80 ℃: 20.09 mg GA/g, 100 ℃ :18.45 mg GA/g). There were high content of flavonoid and rutin as the leaching temperature increased. Consistently, 2,2-diphenyl1-picrylhydrazyl (DPPH) radical scavenging activity and tyrosinase inhibitory activity were significantly higher with increasing temperature (DPPH % inhibition: 60 ℃: 41.88%, 80 ℃: 46.01%, 100 ℃: 46.80%/tyrosinase inhibitory activity: 60 ℃: 9.38%, 80 ℃: 22.94%, 100 ℃: 28.17%). However, there was no significant difference in DPPH radical scavenging activity between 80 and 100 ℃. A cytotoxicity test was performed by treating with Jeju Tatary buckwheat extract into mouse macrophage cells (Raw264.7). 100 and 200 ㎍/mL treatment (100 ℃ extract) were significantly upregulated the survival rate, but there was no significant difference in other concentrations. Collectively, most of the bioactive components, antioxidant activity, and tyrosinase inhibitory activity were induced as the leaching temperature increased. However, the content of polyphenols which are known to have antioxidant activity, was significantly reduced at 100 ℃ leaching temperature. Several reports have demonstrated that leaching at too high temperature lowered the overall acceptability, so the optimal leaching condition of Tatary Buckwheat is 80 ℃, 5 min in this study.