• Title/Summary/Keyword: 대식세포주

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Evaluation of Antioxidant and Anti-inflammatory Activities of Stachys Sieboldii MIQ. and Helianthus tuberosus L. of Cultivated in Sunchang-gun (순창산 초석잠과 돼지감자의 항산화 활성 및 항염증 활성 평가)

  • Se-Won Lee;Mina Sin;Tae-Hu Jang;Hyun-Jin Tae;Dae-Geun Kim;Kyeon-Ok Jeong;Jeong-Ho Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.84-84
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    • 2020
  • 염증 (Inflammation)은 병원체, 손상된 세포, 자극물질 등으로 인한 손상에 대해 작용하여 조직이나 장기의 손상을 재생하는 작용으로써 신체 방어 기전들 가운데 매우 중요한 역할을 하는 것으로 알려져 있다. 이러한 염증반응이 과다할 경우 각종 염증성 질환 혹은 암 등을 유발하는 원인으로 발전할 수 있어 항염증제의 개발은 전 세계적으로 중요시되고 있다. 선천적 면역을 담당하는 대식세포는 lipopolysaccharide(LPS), 활성산소 (ROS) 등에 의해 자극되어 염증인자 생성에 관여한다. 따라서 본 연구에서는 항염증 소재 개발을 목표로 하며, 국내 천연소재를 활용한 기능성 항염증 소재로 전북 순창군에서 재배된 초석잠(Stachys Sieboldii MIQ.)과 돼지감자 (Helianthus tuberosus L.)를 이용하여 항산화 활성을 평가하고자 하였으며, 대식세포주를 활용한 세포독성 및 항염증 활성에 대한 효능을 확인하고자 하였다. 본 연구는, 전북 순창 지역에서 재배 된 초석잠과 돼지감자를 사용하여 각 조건의 추출 용매, 온도, 시간별 추출물의 Total polyphenol 함량 평가를 통한 최적 추출조건 선정을 진행하고, 선정된 추출조건의 추출물의 항산화 활성을 측정하기 위해 DPPH & ABTS radical scavenging activity 분석 및 Total flavonoids 함량 분석을 통해 항산화 효능 평가를 진행하였다. 또한 항염증 소재로의 활용을 위해 대식세포인 Raw 264.7을 사용하여 농도별 MTT assay를 진행하여 세포독성 평가를 진행하였고, Nitric oxide (NO) 생성억제 효능을 확인하여 항염증 활성을 평가하였다. 실험 결과, Total polyphenol 함량 분석을 통해 최적 추출조건이 선정된 초석잠 (25℃, 주정 60%, 3 h), 돼지감자 (25℃, 주정 40%, 1 h)은 최적 조건에서 약 58 mg GAE/g 및 158 mg GAE/g의 총 폴리페놀 함량을 보였으며, DPPH & ABTS radical scavenging activity 및 Total flavonoids 함량 분석한 결과, 초석잠이 돼지감자보다 더 높은 항산화 활성을 나타내었다. 대식세포 실험에서의 추출물 처리군의 세포독성 측정 결과, 100 ug/mL 이내의 농도에서 독성활성이 나타나지 않음을 확인하였고, Nitric oxide (NO) 생성 억제활성 측정을 통해 LPS 처리군 대비 접종량 100 ug/mL 기준 각각 초석잠 약 47%, 돼지감자 약 49% 수준의 항염증 활성을 확인하였다.

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Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

Inhibition of lipopolysaccharide-stimulated inflammatory mediator production in RAW264.7 macrophages by Foeniculum vulgare fruit extract (LPS로 자극한 RAW264.7 대식세포주에서 회향 추출물에 의한 염증성 매개물의 생성 억제)

  • Choi Eun-Mi;Koo Sung-Ja
    • Korean journal of food and cookery science
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    • v.20 no.5
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    • pp.505-510
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    • 2004
  • 이물질 침입에 대한 인식의 결과 NO, PGE$_2$, TNF-, IL-6와 같은 여러 신호전달물질의 분비가 개시되며 이들을 억제하는 물질을 항염증제라고 볼 수 있다. 본 연구에서는 회향(Foeniculum vulgare Mill.) 열매 추출물이 mouse macrophages RAW264.7 세포에서 lipopolysaccharide(LPS)로 유도한 NO(iNOS 산물), PGE$_2$(COX-2 산물) 및 cytokines (TNF-$\alpha$, IL-6) 생성 억제에 미치는 영향을 살펴보았다. 회향 열매의 methanol 추출물 및 분획물(chloroform, butanol, and aqueous fractions)은 4~100$\mu$g/mL 농도에서 LPS가 활성화된 대식 세포에서 NO 생성을 억제하였으며 독성을 나타내지 않았다. LPS가 유도한 PGE$_2$ 생성은 butanol 분획(100 $\mu$g/mL)에 의해서만 유의적으로 감소하였다(P<0.05). 회향 열매 추출물 및 분획물은 TNF-$\alpha$의 생성을 유의적으로 감소시켰으며 IL-6의 생성은 methanol extract(4~100 $\mu$g/mL), chloroform fraction(4 $\mu$g/mL), butanol fraction(4 and 100$\mu$g/mL) 및 aqueous fraction(4~100 $\mu$g/mL)에 의해 감소되었다(P<0.05). 이는 회향 열매 추출물은 염증 상태에서 유용할 것이며 COX-2와 iNOS를 억제하는 butanol 분획은 새로운 항염증제 개발에 사용될 수 있음을 시사하여 주었다.

Inhibitory Effect of Taraxacum mongolicums (蒲公英) on NO Production in LPS-stimulated Macrophages (LPS로 자극된 대식세포에서 포공영의 NO 생성 억제 효과)

  • Ha, Hyeon-Hee;Park, Sun-Young;Ko, Woo-Shin;Jang, Jeong-Su;Kim, Young-Hee
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.3
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    • pp.98-106
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    • 2007
  • 포공영(蒲公英)은 예로부터 청열해독약(淸熱解毒藥)으로 사용되어 왔으며 NO가 염증의 한 요인이기 때문에 포공영의 항염증 작용기작을 밝히기 위하여 LPS로 자극된 대식세포주 RAW264.7 세포에서 포공영 열수 추출물 (AETM)의 NO 생성에 미치는 효과를 조사하였다. 포공영은 NO 생성 및 iNOS 단백질 발현, iNOS mRNA 발현을 저해하였으며, 전사인자인 $NF-kB$의 핵으로의 이동을 억제하였다. 또한 LPS에 의해서 활성화되는 ERK/MAPK 효소의 활성을 현저히 억제하였다. 이 결과들로 보아 포공영의 항염증 작용이 ERK/MAPK 활성 저해 및 $NF-kB$ 활성 저해로 인한 iNOS 발현의 억제 때문인 것으로 사료된다.

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Antibacterial and Intracellular Clearance Effect of Saururus chinensis Baill Water Extract against Salmonella typhimurium (삼백초 수용성 추출물의 Salmonella typhimurium 균에 대한 항균 및 숙주세포 내 사멸 효과 규명)

  • Kim, Dong-Hyeok;Lee, Jin-Ju;Lim, Jeong-Ju;Kim, Dae-Geun;Kim, Gon-sub;Lee, Hu-Jang;Min, Won-gi;Rhee, Man-Hee;Chang, Hong-Hee;Kim, Suk
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.111-119
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    • 2010
  • Salmonella spp. are one of major pathogens for zoonosis in worldwide, and can replicate within host cells and generally cause enterocolitis and foodborne poisoning which represents a considerable public health burden. The present study was designated to investigate the safty for host cells, antibacterial effects of Saururus chinensis Baill water extract (SCWE) on pure culture and infection with Salmonella enterica serovar Typhimurium (S. typhimurium) in murine derived macrophage RAW 264.7 cells. The different treatment of SCWE concentration (1, 10 or $100{\mu}g/ml$) did not show any cytotoxic effect to RAW 264.7 cells for 24 h incubation. In determination of antibacterial activity of SCWE against S. typhimurium, bacterial viability was markedly decreased compared to SCWE-untreated control. In RAW 264.7 cells, SCWE significantly induced morphological change (p<0.05). In infection assay of S. typhimurium in RAW 264.7 cells pretreated with $100{\mu}g/ml$ of SCWE, which are non-cytotoxic concentration, bacterial uptake ability of macrophage was increased corresponding with morphological change, whereas bacterial survival rates within macrophage was markedly reduced comparing to that of SCWE-untreated control. Furthermore, nitric oxide (NO) production in SCWE-treated cells was slightly decreased until 24 h post infection. Taken together, these findings demonstrated that SCWE have the antibacterial activity for S. typhimurium and the protective effects against S. typhimurium infection through activating murine macrophage independent on NO, suggesting that SCWE were beneficial on the disease caused by intracellularly replicating pathogens as a safe alternatives of conventional chemotherapies.

Ultrastructure of Macrophages in BAL of Rat Given Interleukin-1$\alpha$ Intratracheally (인터루킨-1$\alpha$를 기관지 투입 후 나타난 폐세척액에서의 대식세포의 미세구조적 변화)

  • 조현국;이영만;박원학
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.159-166
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    • 1996
  • In order to investigate the recycling of the pulmonary surfactant in association with morphological changes in macrophage after interleukin-1 $\alpha$ (IL-1) induced lung injury, an acute lung injury was induced by instillation of IL-1 into the trachea. Numbers of neutrophils and phospholipid content were increased significantly(P<0.01) in IL-1 treated BAL(brochoalveolar lavage) compared to control rat. By increased phagocytosis, the lamellar structures in the macrophges of IL-1 treated rats' BAL were increased and the compositions of the cellular organelles were changed in comparison to control rat. This difference in compositions of cellular organelles denotes difference of functions in macrophages between control and IL-1 treated rats. As macrophages have been said to implicate (in the difference in the recycling of pulmonary surfactant, it is highly probable that the difference in compositions of cellular organelles is closely related to the recycling of pulmonary surfactant. In the present study circular structures were synthesized in the cytoplasm of the macrophages in BAL of normal rats. Based on these experimental results, it is suggested that macrohages might synthesize during recycling of surfacuant in the lung.

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Effects of Aerobic Exercise and Chrysin Supplementation on Macrophage Infiltration and Lipolysis Genes of High-Fat Diet Mice (고지방식이 동물모델에서 크리신 섭취와 유산소 운동이 대식세포 침윤과 지방분해 유전자들에 미치는 영향)

  • Choi, Do-Yourl;Lee, Young-Ran
    • Journal of Digital Convergence
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    • v.17 no.5
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    • pp.399-405
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    • 2019
  • This study was investigated the effect of aerobic exercise and chrysin supplementation on macrophage infiltration and lipolysis in high-fat diet mice. To accomplish the purpose of this study, C57BL/6 mice were fed high fat diet(60% fat diet) during experimental period. The animals were divided into 4 groups; NC (normal diet control, n=5), HC (high fat diet control, n=5), Hch(high fat diet with chrysin, n=5), and HME (high fat diet with aerobic exercise training, n=5). Exercise training was performed for 16 weeks on a treadmill running. As a result, macrophage marker, F480 and CD11c were significantly decreased in HME comparison with HD and Hch. Also, M2 macrophage marker CD11c, and lipolysis marker PRDM were significantly increased in HME compared with HC and Hch These findings suggest that regular aerobic exercise has beneficial effects to inhibit macrophage infiltration in high fat diet mice.

Effect of Arabinoxylane and PSP on Activation of Immune Cells (Arabinoxylane과 PSP의 면역세포 활성화 효과)

  • 채수연;신성해;배만종;박미현;송미경;황성주;이성태
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.2
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    • pp.278-286
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    • 2004
  • The objective of the current study was to determine the effects of arabinoxylane and PSP on mouse splenocytes, T cells, B cells and macrophages in vitro. Arabinoxylane and PSP directly induced the proliferation of spleen cells in a dose-dependent manner and increased IFN-${\gamma}$ synthesis. Especially, PSP induced IL-2, IL-4 and IL-10 production, Both arabinoxylane and PSP increased PFC (plaque forming cell) and RFC (rosette forming cell) formation. Arabinoxylane was not induced the proliferation of T cells, but PSP directly induced the proliferation of T cells in a high dose. Arabinoxylane and PSP increased the proliferation of B cells and the phagocytic effects of macrophage. When arabinoxylane and PSP were used in macrophage cell line stimulation, there was a marked induction of NO synthesis in a dose-dependent and an increased TNF-$\alpha$ and IL-6 synthesis. Especially, PSP also induced IL-1$\beta$ production. When arabinoxylane and PSP treated in macrophage cell line, there was induction of MHC class II expression. These results suggest that the capacity of arabinoxylane andPSP seem to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, arabinoxylane and PSP could be used as a biological response modifier having possible therapeutic effects against immunological disorders.

In vitro Studies on the Superoxide Scavenging Activities, the Cytotoxic and the Immunomodulating Effects of Thirteen Kinds of Herbal Extracts (13종 허브추출물의 수퍼옥사이드 소거능과 세포독성 및 면역증강 효과의 in vitro계 검색)

  • Chung, Ha-Yull;Kim, Hyun-Bae
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.699-705
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    • 2000
  • The superoxide scavenging activities of thirteen kinds of herbal extracts were examined together with their cytotoxic and immunomodulating effects. The extracts of 6 kinds of herbs such as eucalyptus, mate, peppermint, sage, thyme and yarrow, so called medicinal herbs, showed above 70% of the superoxide scavenging activities. They also showed cytotoxicities against the cancer cell lines of Hepa-1c1c7 and KB-3-1 as well as the normal fibroblast cell line of mouse. The $IC_{50}$ values of above 6 herb extracts on the cancer cell lines were above or similar to the $IC_{50}$ values on the normal cell line, so it was unable to observe the herb extract which showed cytotoxicity only to the cancer cell lines. Considering the results of nitric oxide test that the sage extract was the only one having the immunomodulating effect(37% of positive control), there was no significant relationship between the superoxide scavenging activities of herbs and their immunomodulating effects.

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