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Anti-inflammation effect of blueberry (Vaccinium ashei) leaf extract on RAW 264.7 macrophages stimulated by lipopolysaccharide (Lipopolysaccharide에 의해 활성화된 RAW 264.7대식세포에서 블루베리 잎(Vaccinium ashei) 추출물의 항염증 효과)

  • Kim, Dong In;Kim, Hyun Jung;Yun, Jong Moon;Lee, Ji Hye;Han, So Jung;Kim, Ha Eun;Jang, Min Jung;An, Bong Jeun
    • 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.

Production of Antimicrobial Compounds and Cloning of a dctA Gene Related Uptake of Organic Acids from a Biocontrol Bacterium Pseudomonas Chlororaphis O6 (생물적 방제균 Pseudomonas chlororaphis O6의 길항 물질 생산 및 유기산 흡수에 관련된 dctA 유전자의 클로닝)

  • Han, Song-Hee;Nam, Hyo-Song;Kang, Beom-Ryong;Kim, Kil-Yong;Koo, Bon-Sung;Cho, Baik-Ho;Kim, Young-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.3
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    • pp.134-144
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    • 2003
  • A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

Effects of Dietary Supplementation of Mulberry Leaves and Silkworm Excreta Extracts on Weight Performance, Blood Characteristics and Cecal Microflorae of Chicks (뽕잎과 잠분 추출물 급여가 육계의 생산성, 혈액 성상 및 맹장 내 미생물군에 미치는 영향)

  • Han, Sang-Hoon;Kim, Dong Wook;Ji, Sang Yoon;Hong, Seong Koo;Kim, Sang-Ho;Lee, Heui-Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.150-160
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    • 2012
  • The object of this study was to evaluate the effects of dietary supplementation of mulberry leaves and silkworm excreta ethanol extracts on weight performances, blood characteristics, cecal microflorae of chickens. Two hundred forty male broiler chicks(Ross) were fed diets for five weeks containing 0.1%(MLA) and 1%(MLB) of mulberry leaves ethanol extracts, and 0.1%(SEA) and 1%(SEB) of silkworm excreta ethanol extracts. Weight performance did show no significant difference in all test groups which were fed with supplementation of mulberry leaves and silkworm excreta ethanol extracts. They showed better weight gain and feed conversion than the negative control group which was fed only with forage without antibiotics. ABTS(2'-azine-bis[3-ethylbenzothiazoline-6-sulfonic acid]) test was conducted to investigate free radical scavenging activity of blood in tested groups. ABTS scavenging activities of tested groups were higher than control groups in significant level, though there was no significant difference(P = 0.396). Specifically, MLB group showed the highest scavenging activity. Blood-level concentration of MDA, which is an indicator of lipid peroxidation, was also decreased in tested groups and the lowest level was observed in SEA(P = 0.001). As storage time increased at $4^{\circ}C$, muscle-level MDA concentrations of all tested groups were generally increased and significant difference was obsereved between tested groups and controls in total increase of MDA concentration($P=4.417{\times}10^{-3}$). In cecal microflorae, SEA and SEB showed decreased total microbe population compared to NC($P=6.462{\times}10^{-5}$) and even to PC. Supplementation of mulberry leave and silkworm excreta ethanol extract did show a similar inhibition effect against Salmonella sp., furthermore, MLB did enhanced the growth of Lactobacillus sp.($P=3.636{\times}10^{-7}$). In summary, ethanol extract of silkworm excreta may be a potential alternative of antibiotics for chicks. In addition, both of ethanol extracts supplementation to broiler chicks would be very useful not only to improve antioxidant effect of blood but also to suppress lipid peroxidation without any loss of weight performance in poultry farming.

Anti-inflammatory Activities of Antimicrobial Peptide Locustacin Derived from Locusta migratoria in LPS-stimulated RAW264.7 Cells (풀무치 유래 항균 펩타이드 locustacin의 항염증 활성)

  • Choi, Ra-Yeong;Lee, Joon Ha;Seo, Minchul;Kim, In-Woo;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.31 no.10
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    • pp.898-904
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    • 2021
  • Locusta migratoria is a widespread locust species in many parts of the world and is considered an alternative source for the production of protein for value-added ingredients. We previously identified putative antimicrobial peptides derived from L. migratoria through an in silico analysis of its transcriptome. However, its anti-inflammatory effect has not been studied. In this study, we investigated the anti-inflammatory activities of the antimicrobial peptide locustacin (KTHILSFFPSFLPLFLKK-NH2) derived from L. migratoria on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Locustacin (50, 100, and 200 ㎍/ml) significantly reduced the production of nitric oxide (NO) in LPS-stimulated macrophages without any cytotoxicity. Locustacin also inhibited the mRNA and protein expression of pro-inflammatory mediators, such as inducible NO synthase and cyclooxygenase-2, in contrast to the presence of LPS alone. Locustacin decreased the release of LPS-induced pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, and their gene expression in a dose-dependent manner. Furthermore, locustacin (100 and/or 200 ㎍/ml) inhibited phosphorylation levels of extracellular signal regulated kinase, p38, and c-Jun N-terminal kinase. Locustacin also suppressed the degradation of inhibitory kappa B alpha, which was considered to be an inhibitor of nuclear factor kappa B (NF-κB). Collectively, these results demonstrate that locustacin can exert anti-inflammatory effects through the inhibition of mitogen-activated protein kinase (MAPK) phosphorylation, activation of NF-κB, and downstream inflammatory mediators in LPS-stimulated macrophage cells.

Effects of Foliar Application of CaCl2 on Ca Content and Occurrence and Alleviation of Blossom-end Rot of Paprika During High Temperature Season (고온기 CaCl2 엽면 살포가 파프리카 칼슘함량, 배꼽썩음과 발생과 경감에 미치는 영향)

  • Oh, Jeong Sim;Lee, Yong-Beom;Bae, Jong Hyang;Na, Jong Kuk;Choi, Ki Young
    • Journal of Bio-Environment Control
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    • v.30 no.4
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    • pp.263-270
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    • 2021
  • This experiment was carried out to determine the effect of CaCl2 foliar spraying on the inhibition of blossom-end rot (BER) in hot summer paprika (Capsicum annum L. 'Special') cultivation. The effect of CaCl2 application was examined by the foliar application based on different fruit size, frequency, and spraying time. Also, it was investigated the occurrence and alleviation effect of BER symptom. Foliar application of CaCl2 (Ca 0.4%) was conducted by treating a fruit and leaf at 4 to 9 nodes above the crown flowers of each plant with 350 mL per week from June 3 to July 1. When the CaCl2 was sprayed at 7-day intervals for 4 weeks, the Ca content was the lowest in the fruit harvested with BER symptom in 11 to 20 mm of fruit width (FW). Four different regions in both BER symptom and normal fruits showed significant differences of Ca content, the highest was in pedicel, followed by stem-end, middle, and blossom-end. The Ca content increased sharply in normal paprika with 31-40 mm FW, in which Ca content was 78% higher than that of BER. Ca content in the middle and blossom-end of paprika over 21 mm in FW ranged 19.8% to 28.8% in normal fruits and 15.7% to 18.5% in BER, respectively. The incidence of BER increased rapidly by more than 60% in fruits with 31-40 mm FW. While there was no difference in fruit weight among the FW treatment, marketable yield rate was highest in the 21-30 mm FW, and the sugar content was high in the 11-30mm FW. When CaCl2 was applied three times for 7 days to a paprika having a 21- 30 mm FW, the cell wall-bound (CWB) Ca content was the highest and the rate of BER was lowest with 6.3%. After 10 days of CaCl2 foliar spray treatment, the CWB Ca content of paprika increased by 2.9 to 3.5 times compared to the control in all treatments. At 7 days after the CaCl2 foliar spraying once a day at varying spraying time, the leaf burn observed from 9:30 a.m. to 17:00 p.m. and the proline content increased as the spraying time was delayed. Therefore, the CaCl2 foliar spraying method for reducing of BER occurrence during paprika summer cultivation seems to be appropriate to spray 2-3 times at intervals of 3 days and before 8 a.m. at the time when the FW is 21-30 mm.

Comparison of Measured and Calculated Carboxylation Rate, Electron Transfer Rate and Photosynthesis Rate Response to Different Light Intensity and Leaf Temperature in Semi-closed Greenhouse with Carbon Dioxide Fertilization for Tomato Cultivation (반밀폐형 온실 내에서 탄산가스 시비에 따른 광강도와 엽온에 반응한 토마토 잎의 최대 카복실화율, 전자전달율 및 광합성율 실측값과 모델링 방정식에 의한 예측값의 비교)

  • Choi, Eun-Young;Jeong, Young-Ae;An, Seung-Hyun;Jang, Dong-Cheol;Kim, Dae-Hyun;Lee, Dong-Soo;Kwon, Jin-Kyung;Woo, Young-Hoe
    • Journal of Bio-Environment Control
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    • v.30 no.4
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    • pp.401-409
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    • 2021
  • This study aimed to estimate the photosynthetic capacity of tomato plants grown in a semi-closed greenhouse using temperature response models of plant photosynthesis by calculating the ribulose 1,5-bisphosphate carboxylase/oxygenase maximum carboxylation rate (Vcmax), maximum electron transport rate (Jmax), thermal breakdown (high-temperature inhibition), and leaf respiration to predict the optimal conditions of the CO2-controlled greenhouse, for maximizing the photosynthetic rate. Gas exchange measurements for the A-Ci curve response to CO2 level with different light intensities {PAR (Photosynthetically Active Radiation) 200µmol·m-2·s-1 to 1500µmol·m-2·s-1} and leaf temperatures (20℃ to 35℃) were conducted with a portable infrared gas analyzer system. Arrhenius function, net CO2 assimilation (An), thermal breakdown, and daylight leaf respiration (Rd) were also calculated using the modeling equation. Estimated Jmax, An, Arrhenius function value, and thermal breakdown decreased in response to increased leaf temperature (> 30℃), and the optimum leaf temperature for the estimated Jmax was 30℃. The CO2 saturation point of the fifth leaf from the apical region was reached at 600ppm for 200 and 400µmol·m-2·s-1 of PAR, at 800ppm for 600 and 800µmol·m-2·s-1 of PAR, at 1000ppm for 1000µmol of PAR, and at 1500ppm for 1200 and 1500µmol·m-2·s-1 of PAR levels. The results suggest that the optimal conditions of CO2 concentration can be determined, using the photosynthetic model equation, to improve the photosynthetic rates of fruit vegetables grown in greenhouses.

Effects of Schisandrae Fructus and Corni Fructus Extracts on the Proliferation and Expression of Prostatic Hyperplasia-inducing Factors in Dihydrotestosterone-stimulated LNCaP Human Prostate Carcinoma Cells (오미자와 산수유 추출물이 dihydrotestosterone가 처리된 LNCaP 인간 전립선 암세포의 증식 및 전립선 비대 유발 인자 발현에 미치는 영향)

  • Kim, Min Yeung;Ji, Seon Yeong;Hwangbo, Hyun;Lee, Hyesook;Kim, Tae Hee;Yoon, Seonhye;Kim, Hyun Jin;Kim, Sung Yeon;Kim, Tae Jung;Kim, Min Ji;Jung, Ha Eun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.31 no.10
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    • pp.885-897
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    • 2021
  • Schisandrae Fructus (SF) and Corni Fructus (CF) have been used for a long time for the prevention and treatment of various diseases. Although reports have highlighted the possibility of inhibiting the onset and progression of benign prostatic hyperplasia (BPH), studies on related mechanisms are still lacking. In this study, we investigated the potential of SF and CF in improving BPH by using a dihydrotestosterone (DHT)-induced in vitro BPH model using LNCaP prostate carcinoma cells. According to our results, water and ethanol extracts of SF and CF significantly inhibited the proliferation of LNCaP cells by DHT treatment and markedly downregulated the expression of DHT-induced BPH biomarkers and growth factors. They also regulated the expression of apoptosis regulatory factors and significantly reduced DHT-mediated oxidative stress. In addition, the protective effect on major factors involved in the pathogenesis of BPH was more effective in the ethanol extract treatment group than in the water extract group. Furthermore, the improvement effect on BPH was higher in the 1:1 combined treatment group than in the ethanol extract alone treatment group of SF and CF, and 60% ethanol extracts showed a better effect than 40% ethanol extracts. Therefore, our findings demonstrate that SF and CF can protect against BPH by preventing the hyperproliferation of prostate cells through the inhibition of the androgen signaling pathway, which was correlated with their antioxidant activities. Therefore, SF and CF extracts may be useful in the clinical treatment of BPH, and the combination of these two extracts can be synergistic.

Apoptosis Induction by Dendropanax morbiferus Leaves Extract in Human Breast Cancer Cells SK-BR-3 through MAPK Pathway (유방암 세포에서 황칠나무 잎 추출물의 MAPK 경로를 통한 apoptosis 유도)

  • Jung, Gi Hwan;Kim, Sung Hyun;Woo, Joong Seok;Yoo, Eun Seon;Lee, Jae Han;Han, So Hee;Jung, Soo Hyun;Kim, Sae Man;Kim, Eun Gee;Shin, Man Jae;Cho, Ho Min;Jung, Ji Youn
    • Journal of Life Science
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    • v.31 no.9
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    • pp.827-833
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    • 2021
  • Dendropanax morbiferus leaves (DPL) has been used as a medicine since ancient times in various diseases such as inflammation, diabetes, and cancer. In particular, it has been found to have anticancer effects on several types of cancer cells, but the anticancer effect on breast cancer cells SK-BR-3 has not yet been revealed. Therefore, in this study, DPL caused proliferation inhibition in breast cancer cells SK-BR-3 and the anticancer effect by inducing apoptosis was confirmed, through an in vitro experiment. In order to examine the effect of DPL on cell viability, MTT assay was performed to confirm a significant decrease in the concentration of cell viability. DAPI staining was performed to examine the effect of DPL on cellular morphological changes and increase of apoptotic bodies. To supplement this, an increase in the apoptosis rate was also confirmed through flow cytometry after staining with annexin V/PI. Western blot was performed to confirm apoptosis-related proteins. DPL increased the expression of Cleaved-PARP, Bax whereas decreased the expression of Bcl-2. Changes in the expression levels of MAPK pathway proteins p-ERK1/2, p-JNK, and p-p38 were also confirmed, and a significant increase in p-p38 was observed. These results indicated that DPL induced apoptosis, through p-p38 MAPK signal pathway in SK-BR-3 breast cancer cells.

Attenuation of Lipopolysaccharide-induced Inflammatory and Oxidative Response by 5-Aminolevulinic Acid Phosphate in RAW 264.7 Macrophages (RAW 264.7 대식세포에서 lipopolysaccharide 자극에 의한 염증성 및 산화적 스트레스에 미치는 5-aminolevulinic acid phosphate의 영향)

  • Ji, Seon Yeong;Kim, Min Yeong;Hwangbo, Hyun;Lee, Hyesook;Hong, Su Hyun;Cha, Hee-Jae;Kim, Heui-Soo;Kim, Suhkmann;Choi, Yung Hyun
    • Journal of Life Science
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    • v.31 no.9
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    • pp.818-826
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    • 2021
  • 5-Aminolevulinic acid phosphate (5-ALA-p) is a substance obtained by eluting 5-ALA (a natural delta amino acid) with aqueous ammonia, adding phosphoric acid to the eluate, and then adding acetone to confer properties suitable for use in photodynamic therapy applications. However, its pharmacological efficacy, including potential mechanisms of antioxidant and anti-inflammatory reactions, remains unclear. This study aimed to investigate the effects of 5-ALA-p on oxidative and inflammatory stresses in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our data showed that 5-ALA-p significantly inhibited excessive phagocytic activity via LPS and attenuated oxidative stress in LPS-treated RAW 264.7 cells. Furthermore, 5-ALA-p improved mitochondrial biogenesis reduced by LPS, suggesting that 5-ALA-p restores mitochondrial damage caused by LPS. Additionally, 5-ALA-p significantly suppressed the release of nitric oxide (NO) and pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6, which are associated with the inhibition of inducible NO synthase and respective cytokine expression. Furthermore, 5-ALA-p reduced the nuclear translocation of nuclear factor-kappa B (NF-κB) and inhibited phosphorylation of mitogen-activated protein kinases (MAPKs), indicating that the anti-inflammatory effect of 5-ALA-p is mediated through the suppression of NF-κB and MAPK signaling pathways. Based on these results, 5-ALA-p may serve as a potential candidate to reduce inflammation and oxidative stress.

Emodin Studies on Anti-inflammatory and Skin Barrier Improvement Activities (Emodin의 항염 및 피부장벽개선 활성 연구)

  • Kim, Se-Gie;Choi, Jae Gurn;Jang, Young-Ah
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.6
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    • pp.1383-1392
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    • 2021
  • It has been reported that emodin, a major pharmacologically active ingredient of herbal medicines such as Polygonum cuspidatum, Polygonum multiflorum, Rheum palmatum, and Aloe vera, is effective in antioxidant, antibacterial, anti-inflammatory, anticancer, and liver protection. In this study, to investigate the potential of emodin to be used as a skin disease and functional material, the activity related to the improvement of inflammation and skin barrier function was confirmed. To observe the anti-inflammatory effect on HaCaT cells, which are human keratinocytes, cytokine inhibition was confirmed by ELISA kit and protein expression by western blot. In HaCaT cells activated with TNF-α (10 ng/mL)/IFN-γ (10 ng/mL), emodin was treated with each concentration (5, 10, 20, 40) µM. As a result, It was confirmed that the production amount of TNF-α, IL-1β and IL-6 decreased as the concentration of emodin increased. In the experimental results on the expression levels of inflammation-related proteins iNOS and COX-2, it was confirmed that 48% of iNOS and 29% of COX-2 were inhibited compared to control at a concentration of 20 µM of emodin. As an indicator of skin barrier function improvement, the mRNA expression level of filaggrin, involucrin, and loricirn and the production amount of filaggrin, involucrin, and loricirn were confirmed. and excellent results were obtained with an emodin concentration-dependent increase. In particular, filaggrin, which was produced twice as much as the control at a concentration of 20 µM, is a protein involved in the formation of NMF, a natural moisturizing factor, and is known to play an important role in moisturizing the stratum corneum. In conclusion, it was confirmed that emodin can be used as a material for improving inflammation and improving skin barrier function, which is part of the potential for use as a skin disease and functional material. It is believed that if additional research is performed in the future, the scope of its application can be further expanded.