• Title/Summary/Keyword: Macrophage cell

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Anti-Inflammatory Effects of Vegetable Soup in Murine Macrophage RAW 264.7 Cells (야채스프의 RAW 264.7 세포에서 항염증 효과)

  • Sim, Jae-Geun;Lee, Jae-Hyeok;Shin, Tae-Yong;Shin, Hye-Young;Jeong, Sook-Hee;Kim, Mi-Hye;Ku, Ho-Jun;Park, Jeong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1097-1101
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    • 2010
  • The aim of the present study is to investigate the anti-inflammatory effect of a vegetable soup (VS). The present study was designed to determine the effect of the vegetable soup on pro-inflammatory factors such as NO, iNOS and TNF-$\alpha$ in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cell toxicity was determined by MTS assay. To evaluate the anti-inflammatory effect of vegetable soup, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Also, proinflammatory cytokines were measured by ELISA kit. The results showed that the vegetable soup reduced NO, iNOS and TNF-$\alpha$ production without cytotoxicity. Our results suggest that the vegetable soup may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful to develop the functional food realted to anti-inflammation.

Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells

  • Lee, Min-Jung;Kim, Dae-Won;Lee, Yeom-Pyo;Jeong, Hoon-Jae;Kang, Hye-Won;Shin, Min-Jae;Sohn, Eun-Jeong;Kim, Mi-Jin;Jang, Sang-Ho;Kang, Tae-Cheon;Won, Moo-Ho;Min, Bon-Hong;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.42 no.5
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    • pp.286-292
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    • 2009
  • Arginine deiminase (ADI), an arginine-degrading enzyme, has anti-proliferative and anti-tumor activities and is capable of inhibiting the production of nitric oxide (NO). Modulation of nitric oxide (NO) production is considered a promising approach for the treatment of various diseases including cancer, inflammation and neuronal disorders. In this study, an ADI gene was fused with an HIV-1 Tat peptide in a bacterial expression vector to produce an genetic in-frame Tat-ADI fusion protein. When added exogenously to the culture media, the expressed and purified Tat-ADI fusion proteins were efficiently transduced into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced Tat-ADI fusion proteins markedly increased cell viability in cells treated with lipopolysaccharide (LPS). This increase in viability was mediated by an inhibition of NO production. These results suggest that this Tat-ADI fusion protein can be used in protein therapies of NO-related disorders such as cancer, inflammation and neuronal diseases.

Development of High-specificity Antibodies against Renal Urate Transporters Using Genetic Immunization

  • Xu, Guoshuang;Chen, Xiangmei;Wu, Di;Shi, Suozhu;Wang, Jianzhong;Ding, Rui;Hong, Quan;Feng, Zhe;Lin, Shupeng;Lu, Yang
    • BMB Reports
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    • v.39 no.6
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    • pp.696-702
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    • 2006
  • Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/macrophage colony-stimulating factor and FMS-like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno-fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters.

Enhancement of Anti-inflammation Effect by Fermentation Process in Aronia melanocarpa (Michx.) Elliott Extract (발효 공정을 통한 아로니아 추출물의 항염증 효능 증진)

  • Kim, Nam Young;Lee, Young Duck;Cho, Seok Cheol;Shin, Yun Chul;Lee, Hyeon Yong
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.6
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    • pp.475-482
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    • 2014
  • This study was indicated to enhance the anti-inflammation activities by the fermentation of the fruits of Aronia melanocarpa (Michx.) Elliott. The extracts by 70% ethanol (EE) showed better biological activities than those by hot water (WE) from campared result of the effect of extraction solvents. Then, the extract from 70% ethanol extraction was further fermented by lactic acid, denoted as FEE. For antioxidant activities, the FEE had showed the highest value as 0.832 of reducing powder, in comparison with those of EE and WE. Cytotoxicity of the water extraction (WE) was measured for 12.06% in addition of $1.0mg/m{\ell}$ of FEE. For anti-inflammation activities, NO production from the macrophage, RAW 264.7 was observed as $7.24{\mu}M$ and $8.52{\mu}M$ from FEE and EE, respectively. Prostaglandin $E_2$ ($PGE_2$) production from human fibroblast cell, CCD-986sk, was also estimated for $152pg/m{\ell}$ in addition of $1.0mg/m{\ell}$ of the FEE. The lowest production of both IL-6 and TNF-${\alpha}$ were $3.5pg/m{\ell}$ and $865.5pg/m{\ell}$, respectively in addition of $1.0mg/m{\ell}$ of the FEE, whereas $74.5pg/m{\ell}$ and $982.4pg/m{\ell}$ in treated with same concenrations of the EE. It was also found that the FEE was higher amounts than other extracts through HPLC analysis of the anthocyanins. These results strongly indicate that fermentation process of the lactic acid could enhance anti-inflammation activities of extracts by increasing the amounts of the anthocyanins, especially cyanidin-galactoside. Our results suggest that the application of the fermentation process for other medicinal herbs can be improved their biological activities.

Physiological Activities of Extracts of Cedrela sinensis leaves (참죽나무 잎 추출물의 생리활성)

  • Shin, Hee-June;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.23 no.2
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    • pp.164-168
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    • 2008
  • The purpose of this study was to confirm the content of total polyphenol, antioxidative and immune activities of the extracts of Cedrela sinensis leaf. The content of total polyphenol of water extracts ranged from 46.5-59.6 mg/100 g, which was higher than other extracts using organic solvents such as EtOAc, $CH_2Cl_2$ and $C_6H_{14}$. The antioxidant activity of the water and organic solvents extracts showed 6-33% in terms of 2,2-diphenyl-picryl-hydrazyl (DPPH) scavenging activity. To analyze the immuno-stimulation activity of C. sinensis leaf extract, we investigated the effect of the extracts on NO synthesis which is important in host defense against bacterial infection. Hot water extracts significantly increased NO generation by RAW 264.7, macrophage cell line, while organic solvent extract has no significant effect on NO production. To further analyzed the anti-inflammatory effect of the extracts, we investigated the effects of the extracts on lipopolysaccharide(LPS)-induced NO generation. Organic solvent extracts of C. sinensis leaves showed strong inhibitory effect on NO production in LPS-stimulated RAW 264.7 cells. These results suggest that C. sinensis leaf extract may represent a useful immune stimulating agent and anti-inflammatory agent.

Antioxidant Activity and Physiological Function of the Anomala albopilosa Extracts (청동풍탱이(Anomala albopilosa)추출물의 항산화성 및 생리기능)

  • Yoon, Weon-Jong;Lee, Jung-A;Kim, Ji-Young;Kim, Sang-Bum;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.670-677
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    • 2007
  • We analyzed antioxidant and physiological activities to investigate the functional effects of ethanol extracts of Anomala albopilosa imago and A. albopilosa larva. In order to effectively screen for anti-inflammatory agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, xanthine oxidase inhibitory activity, and nitric oxide scavenging capacity of the A. albopilosa extracts. By the screening system, we found that A. albopilosa extracts had antioxidant activity which increased with increments of the extract concentration. Moreover, we examined the inhibitory effect of the A. albopilosa extracts on the production of anti-inflammatory factors that the nitric oxide (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and prostaglandin $E_2\;(PGE_2)$ production activated with LPS ($1{\mu}g/mL$) in murine macrophage cell line RAW 264.7. A. albopilosa extracts potentially inhibited the iNOS and COX-2 in a dose-dependent manner. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. Additionally, the $PEG_2$ production is markedly inhibited after a treatment with the A. albopilosa extracts.

ORF5a Protein of Porcine Reproductive and Respiratory Syndrome Virus is Indispensable for Virus Replication (PRRS 바이러스 ORF5a 단백질의기능학적역할)

  • Oh, Jongsuk;Lee, Changhee
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.1-8
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    • 2015
  • In this study, a DNA-launched reverse genetics system was developed from a type 2 porcine reproductive and respiratory syndrome virus (PRRSV) strain, KNU-12. The complete genome of 15,412 nucleotides was assembled as a single cDNA clone and placed under the eukaryotic CMV promoter. Upon transfection of BHK-tailless pCD163 cells with a full-length cDNA clone, viable and infectious type 2 progeny PRRSV were rescued. The reconstituted virus was found to maintain growth properties similar to those of the parental virus in porcine alveolar macrophage (PAM) cells. With the availability of this type 2 PRRSV infectious clone, we first explored the biological relevance of ORF5a in the PRRSV replication cycle. Therefore, we used a PRRSV reverse genetics system to generate an ORF5a knockout mutant clone by changing the ORF5a translation start codon and introducing a stop codon at the 7th codon of ORF5a. The ORF5a knockout mutant was found to exhibit a lack of infectivity in both BHK-tailless pCD163 and PAM-pCD163 cells, suggesting that inactivation of ORF5a expression is lethal for infectious virus production. In order to restore the ORF5a gene-deleted PRRSV, complementing cell lines were established to stably express the ORF5a protein of PRRSV. ORF5a-expressing cells were capable of supporting the production of the replicationdefective virus, indicating complementation of the impaired ORF5a gene function of PRRSV in trans.

The Inhibitory Effects of Sabaek-san and Sabaeksan plus Sasam on the IL-6, IL-8 and GM-CSF mRNA Levels in Human Epithelial Cells (사백산(瀉白散)과 사백산가사삼(瀉白散加沙蔘)이 인간 기관지상피세포의 IL-6, IL-8 및 GM-CSF 발현에 미치는 영향)

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Chung, Kwang-Jin
    • The Journal of Internal Korean Medicine
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    • v.23 no.1
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    • pp.15-23
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    • 2002
  • Objective : We aimed to identify the dose-dependent inhibitory effects of Sabaek-san(瀉白散) and Sabaeksan plus Sasam(Adenophorae Radix) 瀉白散加沙蔘) on the mRNA expressions of Interleukin(IL)-6, IL-8 and granulocyte macrophage colony stimulating factor(GM-CSF) involved in the asthma model. Materials and Methods : Through this study, BEAS-2B cell lines, human epithelial cells were used. These cells were stimulated by tumor necrosis factor(TNF)-${\alpha}$, IL-1${\beta}$ and histamine for artificial inflammatory expression. ${\beta}$-action messenger RNA(mRNA) was used for standards. After each 24hours of Sabaeksan and Sabaeksan plus Sasam treatment, total cellular RNAs were collected by treating RNA zol directly on living cells, Then the transcriptional activities of IL-6, 8 and GM-CSF were measured by RT-PCR with electrophoresis, Results : The mRNA expressions of IL-6 are significantly inhibited compared to those of controlled group at 40 and 100ug/ml of Sabaeksan extract and $100{\mu}g/ml$ of Sabaeksan plus Sasam extract (p<0.05). The mRNA expressions of IL-8 are significantly inhibited compared to that of controlled group at 2.40 and 100 ug/ml of Sabaeksan extract and $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract(p<0.05) THe mRNA expressions of GM-CSF are significantly inhibited compared to those of the controlled group at $100{\mu}g/ml$ of Sabaeksan extract adn $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract.(p<0.05) Conclusions : This study shows that Sabaeksan and Sabaeksan plus Sasam have dose-dependent inhibitory effects on the mRNA expressions of IL-6, IL-8 and GM-CSF in human epithelial cells. Therefore, these types of herb medicine may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanisms of inhibition by herb medicine in the asthma model.

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Stimulation of the Immune Response by Herbal Formulas for Wind-Cold and Heat Pain Symptom (대표적인 풍한열(風寒熱)에 의한 통증 치료 처방의 면역 활성화 비교 연구)

  • Jung, Da-Young;Ha, Hye-Kyung;Lee, Ho-Young;Lee, Jin-Ah;Lee, Jun-Kyoung;Huang, Dae-Sun;Shin, Hyeun-Kyoo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.4
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    • pp.616-623
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    • 2010
  • Three herbal formulas (Bangpungtongsung-san, Ohyaksungi-san, and Ojeok-san) for wind-cold and heat pain symptom were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, lgG1, and total IgM classes) in plasma were measured. Ohyaksungi-san increased NO synthesis in RAW 264.7 cells. Ojeok-san and Ohyaksungi-san significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Three herbal formulas for wind-cold and heat pain symptom also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. These results suggested that three herbal formulas for wind-cold and heat pain symptom could be used as stimulator of immune response.

Isolation of porcine reproductive and respiratory syndrome virus(PRRSV) in Korea (돼지생식기 및 호흡기증후군(Porcine Reproductive and Respiratory Syndrome ; PRRSV) 바이러스의 국내분리주 작성에 관한 연구)

  • Kweon, Chang-hae;Kwon, Byung-joon;Lee, Han-jung;Cho, Jae-jin;Hwang, Eui-kyung;Shin, Jin-ho;Yoon, Yong-dhuk;Kang, Yung-bai;An, Soo-hwan;Kim, Yong-hee;Huh, Won;Jun, Moo-hyung;Wensvoort, G.
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.77-83
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    • 1994
  • Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.

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