• Title/Summary/Keyword: In vivo

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Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts (괭생이 모자반 추출물의 비장세포 면역활성 증강 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Han, In-Jun;Lee, Byung-Soo;Park, Sang-Yun;Nho, Eun Young;Eom, Ji;Kim, Geon;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.52 no.6
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    • pp.515-528
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    • 2019
  • Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Simultaneous Detection of Seven Phosphoproteins in a Single Lysate Sample during Oocyte Maturation Process (난자성숙 과정의 단일 시료에서 일곱 가지 인산화 단백질의 동시 분석 방법)

  • Yoon, Se-Jin;Kim, Yun-Sun;Kim, Kyeoung-Hwa;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.187-197
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    • 2009
  • Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

Selective Algicidal Effects of a Newly Developed GreenTD against Red Tide Harmful Alga (GreenTD 물질을 이용한 유해 적조 발생 종의 선택적 살조능 평가)

  • Lee, Minji;Shin, Juyong;Kim, Jin Ho;Lim, Young Kyun;Cho, Hoon;Baek, Seung Ho
    • Korean Journal of Environmental Biology
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    • v.36 no.3
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    • pp.359-369
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    • 2018
  • Harmful algal blooms (HABs) are a serious problem for public health and fisheries industries, thus there exists a need to investigate the possible ways for effective control of HABs. In the present study, we investigated the algicidal effects of a newly developed GreenTD against the HABs (Chattonella marina, Heterosigma akashiwo, Cochlodinium polykriokides, and Heterocapsa circularisquama) and non-HABs (Chaetoceros simplex, Skeletonema sp. and Tetraselmis sp.), which is focused on the different population density and concentration gradients of algicidal substances. The time series viability of target alga was assessed based on the activity of Chl. a photosynthetic efficiency in terms of $F_v/F_m$, and in vivo fluorescence (FSU). Effective control of Raphidophyta, C. marina and H. akashiwo was achieved at a GreenTD concentration of $0.5{\mu}gL^{-1}$ and $0.2{\mu}gL^{-1}$, respectively, and regrowth of both the species was not observed even after 14 days. The inhibitory ratio of the dinoflagellate, C. polykriokides was more than 80% at $0.2{\mu}gL^{-1}$ of GreenTD. H. circularisquama was constantly affected in the presence of $0.2{\mu}gL^{-1}$ of GreenTD in the high- and low-population density experimental groups. On the other hand, diatoms, C. simplex, and Skeletonema sp. were not significantly affected even in the presence of $0.2{\mu}gL^{-1}$ of GreenTD and exhibited re-growth activity with the passage of incubation time. In particular, green alga Tetraselmis sp. remained unaffected even in the presence of the highest concentration of GreenTD ($1.0{\mu}gL^{-1}$), implying that non-HABs were not greatly influenced by the algicidal substances. As a result, the algicidal activity of GreenTD on the harmful and nonharmful algae was as follows: raphidophyte>dinoflagellates>diatoms>green alga. Consequently, our results indicate that inoculation of GreenTD substances into natural blooms at a threshold concentration ($0.2{\mu}gL^{-1}$) can maximize the algicidal activity against HABs species. If we consider the dilution and diffusion rate in the field application, it is hypothesized that GreenTD will demonstrate economic efficiency, thus leading to effective control against the target HABs in the closed bay.

A Consideration of Perception on Enforcement of Serious Accident Punishment Act(SAPA) among the Workers in the Nuclear Medicine Department (중대재해처벌법 시행에 따른 핵의학 종사자의 인식 고찰)

  • Lee, Joo-Young
    • Journal of the Korean Society of Radiology
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    • v.16 no.4
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    • pp.477-490
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    • 2022
  • Serious Accident Punishment Act(SAPA) went into effect as of Jan. 27, 2022. The subject of study was the worker of the nuclear medicine department and the investigation was aimed at identifying the present situation of their understanding on the issue in the here and now, which can be utilized as basic research for further study. The survey was conducted on 51 people of the worker in the nuclear medicine department. The general factors were classified by their gender, the scale of the hospitals, the period of career, and the detailed occupational categories. The conclusion was drawn, including 1 missing data in gender and 2 in the type of occupation. The targeted hospitals were tertiary hospital, university hospital, and general hospital which have nuclear medicine department in. The period of subjects' career was categorized by less than 3 years, 3 to 5 years, 5 to 10 years, and more than 10 years. The specific occupation was classified by in-vivo radiological technologist, radiation safety manager and others. The amount of pressure that the job entails was highest in the category of general hospital, the period of 3 to 5 years of job experience, and radiation safety manager each. The system of the code was well constructed in the category of general hospital, the period of less than 3-year career, and radiation safety manager, as they responded. The blood transmissible disease had the largest number of outbreak of accidents related to the serious industrial accident. In addition, the radiopharmaceutical dosing error had the highest number of outbreak of accidents related to the serious civil accident. Therefore, we need to improve SAPA, facility inspection, security of budget, security of professional manpower. It will help the stable use of radiation and ensure patient safety.

Inhibitory Effect of Potato Sprouting Inhibitor Chlorpropham on Dry Rot (감자 맹아억제제 Chlorpropham의 마른썩음병 억제 효과)

  • Kyusuk Han;Byung Sup Kim;Sae Jin Hong;Nam Sook Kim
    • Journal of Bio-Environment Control
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    • v.32 no.2
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    • pp.165-171
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    • 2023
  • Potato dry rot is one of the potato storage diseases caused by Fusarium species and is a representative pathological disorder that induced post-harvest loss during storage. Chlorpropham treatment for sprouting inhibition is mainly used for room temperature storage of potatoes for processing. In this study, the inhibitory effect of chlorpropham on Fusarium-induced dry rot of potato 'Dano'. To investigate the mycelial growth rate of the dry rot fungus (Fusarium solani Appel & Wollenw), mycelial growth was investigated in a chlorpropham (5.0, 50.4, 503.8, and 5,038 ppm) and prochloraz (0.1, 1.0, 10.0, and 100.0 ppm) medium containing F. oxysporum mycelia. Mycelia were more inhibited as the concentration of chlorpropham and prochloraz increased during incubation at 20℃, and the inhibition rate was 98.2% and 100% when treated with 503.8 ppm of chlorpropham and 10ppm of prochloraz in 14 days, respectively. Potato Dano tubers inoculated with F. oxysporum were dipped in chlorpropham (5.0, 50.4, and 503.8 ppm) and prochloraz (100 ppm) to investigate the effect of preventing dry rot during cold storage at 20℃ and 4℃ in vivo. The disease diameter of potatoes stored at room temperature (about 20℃) was reduced to 13.0 mm in the prochloraz 100 ppm teatment, and 10.7 mm in the chlorpropham 50.4 ppm treatment compared to 13.7 mm in the control tuber at 70 days of storage. The disease progression in all treatments including control was similar with no statistically significant difference at 4℃ air temperature. From the results of this study, it is considered that treatment with 50.4 ppm of chlorpropham after harvest will be useful for suppressing dry rot of stored potatoes.

Effects of polygalacin D extracted from Platycodon grandiflorum on myoblast differentiation and muscle atrophy (길경에서 추출한 polygalacin D가 근원세포 분화 및 근위축에 미치는 영향)

  • Eun-Ju Song;Ji-Won Heo;Jee Hee Jang;Eonmi Kim;Yun Hee Jeong;Min Jung Kim;Sung-Eun Kim
    • Journal of Nutrition and Health
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    • v.56 no.6
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    • pp.602-614
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    • 2023
  • Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.

Ameliorative Effects of Soybean Leaf Extract on Dexamethasone-Induced Muscle Atrophy in C2C12 Myotubes and a C57BL/6 Mouse Model (콩잎 추출물의 근위축 개선 효과)

  • Hye Young Choi;Young-Sool Hah;Yeong Ho Ji;Jun Young Ha;Hwan Hee Bae;Dong Yeol Lee;Won Min Jeong;Dong Kyu Jeong;Jun-Il Yoo;Sang Gon Kim
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1036-1045
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    • 2023
  • Sarcopenia, a condition characterized by the insidious loss of skeletal muscle mass and strength, represents a significant and growing healthcare challenge, impacting the mobility and quality of life of aging populations worldwide. This study investigated the therapeutic potential of soybean leaf extract (SL) for dexamethasone (Dexa)-induced muscle atrophy in vitro and in an in vivo model. In vitro experiments showed that SL significantly alleviated Dexa-induced atrophy in C2C12 myotube cells, as evidenced by preserved myotube morphology, density, and size. Moreover, SL treatment significantly reduced the mRNA and protein levels of muscle RING-finger protein-1 (MuRF1) and muscle atrophy F-box (MAFbx), key factors regulating muscle atrophy. In a Dexa-induced atrophy mouse model, SL administration significantly inhibited Dexa-induced weight loss and muscle wasting, preserving the mass of the gastrocnemius and tibialis anterior muscles. Furthermore, mice treated with SL exhibited significant improvements in muscle function compared to their counterparts suffering from Dexa-induced muscle atrophy, as evidenced by a notable increase in grip strength and extended endurance on treadmill tests. Moreover, SL suppressed the expression of muscle atrophy-related proteins in skeletal muscle, highlighting its protective role against Dexa-induced muscle atrophy. These results suggest that SL has potential as a natural treatment for muscle-wasting conditions, such as sarcopenia.

A prognosis discovering lethal-related genes in plants for target identification and inhibitor design (식물 치사관련 유전자를 이용하는 신규 제초제 작용점 탐색 및 조절물질 개발동향)

  • Hwang, I.T.;Lee, D.H.;Choi, J.S.;Kim, T.J.;Kim, B.T.;Park, Y.S.;Cho, K.Y.
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.1-11
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    • 2001
  • New technologies will have a large impact on the discovery of new herbicide site of action. Genomics, combinatorial chemistry, and bioinformatics help take advantage of serendipity through tile sequencing of huge numbers of genes or the synthesis of large numbers of chemical compounds. There are approximately $10^{30}\;to\;10^{50}$ possible molecules in molecular space of which only a fraction have been synthesized. Combining this potential with having access to 50,000 plant genes in the future elevates tile probability of discovering flew herbicidal site of actions. If 0.1, 1.0 or 10% of total genes in a typical plant are valid for herbicide target, a plant with 50,000 genes would provide about 50, 500, and 5,000 targets, respectively. However, only 11 herbicide targets have been identified and commercialized. The successful design of novel herbicides depends on careful consideration of a number of factors including target enzyme selections and validations, inhibitor designs, and the metabolic fates. Biochemical information can be used to identify enzymes which produce lethal phenotypes. The identification of a lethal target site is an important step to this approach. An examination of the characteristics of known targets provides of crucial insight as to the definition of a lethal target. Recently, antisense RNA suppression of an enzyme translation has been used to determine the genes required for toxicity and offers a strategy for identifying lethal target sites. After the identification of a lethal target, detailed knowledge such as the enzyme kinetics and the protein structure may be used to design potent inhibitors. Various types of inhibitors may be designed for a given enzyme. Strategies for the selection of new enzyme targets giving the desired physiological response upon partial inhibition include identification of chemical leads, lethal mutants and the use of antisense technology. Enzyme inhibitors having agrochemical utility can be categorized into six major groups: ground-state analogues, group specific reagents, affinity labels, suicide substrates, reaction intermediate analogues, and extraneous site inhibitors. In this review, examples of each category, and their advantages and disadvantages, will be discussed. The target identification and construction of a potent inhibitor, in itself, may not lead to develop an effective herbicide. The desired in vivo activity, uptake and translocation, and metabolism of the inhibitor should be studied in detail to assess the full potential of the target. Strategies for delivery of the compound to the target enzyme and avoidance of premature detoxification may include a proherbicidal approach, especially when inhibitors are highly charged or when selective detoxification or activation can be exploited. Utilization of differences in detoxification or activation between weeds and crops may lead to enhance selectivity. Without a full appreciation of each of these facets of herbicide design, the chances for success with the target or enzyme-driven approach are reduced.

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Effects of Ethylacetate Fraction of Plantago asiatica L. on Hypercholesterolemia Induced by High Cholesterol Diet in Rats (질경이(Plantago asiatica L.) 에틸아세테이트분획이 흰쥐의 고콜레스테롤혈증에 미치는 효과)

  • Lee Jae-Joon;Lee Jeong-Hwa;Jeong Chang-Ju;Choi Hyun-Sook;Lee Myung-Yul
    • Food Science and Preservation
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    • v.12 no.6
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    • pp.624-630
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    • 2005
  • Plantain was extracted with ethanol and fractionated systemically with n-hexane, chloroform, ethylacetate, n-butanol and water to study inhibitory effect on cholesterol synthesis in vitro. To screen the effect, inhibitory activities on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase obtained from Saccharomyces cerevisiae were examined using the five fractions of Plantain. The HMG-CoA reductase activity was inhibited most by ehylacetate fraction among the fractions, although the all five fractions had the effect To see the hypocholesterolemic effect of the ethylacetate fraction of Plantanin (PAE) in vivo, male Sprague-Dawley rats were received 5 types of diets for 6 weeks: normal diet group (NOR), high cholesterol diet group($1\%$ cholesterol and $0.25\%$ sodium cholate, CON), normal diet and PAE 70 mg/kg administered group(S1), high cholesterol diet and PAE 140 mg/kg administrated group(S2), and high cholesterol diet and PAE 140 mg/kg administered group(S3). Body weight gains of the CON were significantly increased compared to those of S1, S2 and S3. Activities of serum AST and ALT were tended to be increased in CON compared with NOR and reduced by the PAE administration. Concentrations of serum total cholesterol, free cholesterol, LDL-cholesterol, triglyceride and the atherogenic index were tended to be decreased in the PAE administered groups compared with the CON. HDL-cholesterol and phospholipid concentrations were significantly decreased in the CON and markedly increased by the PAE administered groups. Taten together, it is suggested that the ethylacetate fraction of Plantanin exerts antiatherosclerotic effect by reducing serum cholesterol concentrations in rats fed high cholesterol diets.

Anti-Atopic Effects of Mixed Extracts from Date Plum, Persimmon, and Mulberry Leaves (고욤잎, 감잎 및 뽕잎 복합추출물의 항아토피 효과)

  • Cho, Byoung Ok;Yin, Hong Hua;Shin, Jae Young;Fang, Chong Zhou;Chang, Che Denis;Jang, Seon Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.501-509
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    • 2016
  • The present study investigated the anti-atopic effects of mixed extracts from date plum, persimmon, and mulberry leaves (DPME) on atopic dermatitis (AD)-like skin lesions in hairless mice. The in vivo results demonstrated that DPME treatment significantly reduced the dermatitis clinical score and epidermal thickness in AD-like skin lesions. Histological analyses showed that DPME treatment strongly inhibited dermal infiltration of inflammatory cells and activity of mast cells in AD-like skin lesions. DPME treatment inhibited production of serum IgE and interluekin (IL)-4 in hairless mice with AD. Moreover, DPME treatment significantly suppressed production of tumor necrosis factor $(TNF)-{\alpha}$ and IL-6 cytokines in phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated HMC-1 human mast cells. In addition, DPME treatment reduced production of pro-inflammatory mediators (nitric oxide, prostaglandin E2, $TNF-{\alpha}$, and IL-6) in lipopolysaccharide-stimulated RAW 264.7 macrophages. Therefore, the results of this study indicate that the anti-atopic and anti-inflammatory effects of DPME may be involved in the regulation of inflammatory responses, suggesting that DPME may be used as an anti-atopic dermatitis material and natural anti-inflammatory ingredient.