• Toxicological Research
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• v.24 no.4
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• pp.315-320
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• 2008

Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a glycoprotein and hematopoietic growth factors that regulates the proliferation of myeloid precursor cells and activates mature granulocytes and macrophages. In a previous study, we reported that hGM-CSF could be produced in transgenic rice cell suspension culture, termed rhGM-CSF. In the present study, we examined the repeated dose toxicity of rhGM-CSF in SD rats. The repeated dose toxicity study was performed at each dose of 50 and 200 ${\mu}g/kg$ subcutaneous administration of rhGM-CSF everyday for 28-days period. The results did not show any changes in food and water intake. There were also no significant changes in both body and organ weights between the control and the tested groups. The hematological and blood biochemical parameters were statistically not different in all groups. These results suggest that rhGM-CSF may show no repeated dose toxicity in SD rats under the conditions.

• Korean Journal of Pharmacognosy
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• v.39 no.2
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• pp.104-109
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• 2008

We previously reported that anti-inflammatory properties of poncirin, isolated from fruit of Poncirus trifoliata, might be the result from the inhibition of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis $factor-{\acute{a}}$ ($TNF-{\alpha}$) and interlukin-6 (IL-6) expression via the down-regulation of $NF{-\kappa}B$ binding activity. In this study, we investigated whether poncirin has an inhibitory effect on the production of pro-inflammatory mediators ex vivo and whether poncirin could relieve the symptoms of dextran sulfate sodium (DSS)-induced colitis in mice model of inflammatory bowel disease. Poncirin significantly inhibited the productions of NO, IL-6 and $TNF-{\alpha}$ in lipopolysaccharide (LPS)-induced mouse peritoneal macrophage. In addition, poncirin-treated mice when compared to control mice not receiving treatment recovered better from the weight loss caused by DSS-induced colitis. Changes in disease activity index (DAI) of poncirin-treated mice were also more favorable than for control mice and were comparable with mice treated with a typical anti-inflammatory-drug, 5-aminosalichylic acid (5-ASA). In addition, suppression of plasma NO and IL-6 productions of poncirin-treated mice was also observed in DSS-induced colitis. These results suggest that poncirin has potentially useful anti-inflammatory effects mediated by suppression of inflammatory mediator productions.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.1000-1010
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• 2005

This study was carried out to investigate the comparison of immunomodualtory effects of water-extracted Aconiti lateralis Preparata Radix(PR), Zingiberis Rhizoma(ZR), Cinnamomi Cortex(CC) and Evodiae Fructus(EF). The parameter examined to assess apparent immunomodulatory effect of the water-extracted PR, ZR, CC and EF included the regulation of Nitric oxide (NO). Also, ZR and EF represent the expression of Th1/Th2 type cytokine, the change of B cell phenotype. The water-extracted PR, ZR, CC and EF inhibited NO production and iNOS protein expression in LPS stimulated RAW 264.7 macrophage cells. In the Th1 and Th2 cytokine expression, the water-extracted ZR and EF induced IL-2, IFNr and IL-10 mRNA gene expression. Therefore, it seems that the water-extracted ZR and EF have a inducing effect of Th1 and Th2 type cytokines. In the Flow cytometry analysis, the water-extracted ZR and EF changed B cell phenotype (CD45R/B220), did NOT in PR and CC. The water-extracted PR, ZR, CC and EF have a reducing effect of immune suppression cause by Methotrexate (MTX), an agent of immune suppression. These results suggest that the immunomodulatory effects of the water-extracted ZR and EF may be, in part, associated with the inducing IL-2 and IFNr mRNA gene expression In and regulation of NO production in macrophage cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.3
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• pp.603-608
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• 2006

In activated macrophage, large amounts of nitric oxide (NO) are generated by inducible nitric oxide synthase (iNOS), resulting in acute or chronic inflammatory disorders. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, water extracts from the fruit of Chaenomeles sinensis, the root of Polygonum cuspidatum and Boswellia carterii inhibited the LPS-induced NO production in a parallel dose-dependent manner. To investigate the mechanism by which those extracts inhibits NO production, we examined the expression of iNOS and COX-2 in both mRNA and protein levels. We observed a significant change in the iNOS expression between LPS alone and LPS plus those extracts-treated cells. However, water extracts from Chaenomeles sinensis, Polygonum cuspidatum and Boswellia carterii did not inhibit COX-2 expression which was induced by LPS treatment in Raw 264.7 cells. These data suggest that water extracts from Chaenomeles sinensis, Polygonum cuspidatum and Boswellia carterii can modulate anti-inflammatory immune response, which may be in part associated with the regulation of NO synthesis through the regulation of iNOS expression in mouse macrophage cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.6
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• pp.956-961
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• 2010

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a major mediator of immuno-inflammatory activity. The purpose of this study is to investigate whether TNF-${\alpha}$ productions of mouse macrophage RAW 264.7 and human hepatocyte HepG2 are modulated by Artemisiae argi Folium water extract (AW), Lactobacillus pentosus-fermented Artemisiae argi Folium water extract (AFL), and Saccharomyces cerevisiae-fermented Artemisiae argi Folium water extract (AFS) for 3 h of incubation. Effect of AW on cell viability of HepG2 was also investigated. TNF-${\alpha}$ productions were measured by Enzyme-Linked Immnunosorbent Assay method and cell viability was measured by MTT assay. Both AFL and AFS significantly increased TNF-${\alpha}$ productions of RAW 264.7 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). Also, AFL and AFS significantly increased TNF-${\alpha}$ productions of HepG2 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). AW significantly increased TNF-${\alpha}$ production of HepG2 at the concentration of 100 and 200 ${\mu}g$/mL (p<0.05). AW did not show any cytotoxicity on HepG2 cells for 3 h. These results suggest that AFL, AFS, and AW have the immune-enhancing property related with its increasing effect on TNF-${\alpha}$ production of macrophage and hepatocyte.

• Journal of Korean society of Dental Hygiene
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• v.13 no.1
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• pp.174-180
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• 2013

연구목적 : 자극성 섬유종은 만성자극에 의해 발생하는 구강내 증식성 병변이다. 상처치유의 초기 과정에서는 비만 세포와 대식 세포가 섬유모세포의 이주, 증식, 아교질합성 등에 연관되어 있는 성장인자와 사이토카인을 분비하여 상처 치유에 중요한 역할을 한다. 저자들은 자극성 섬유종을 조직학적 특성에 따라 세분하고, 각각의 조직학적 아형에서 비만 세포와 대식 세포의 발현을 조사하여 자극성 섬유종의 발생 기전을 이해하고자 하였다. 연구방법 : 본 연구에서는 82예의 자극성 섬유종을 조직 소견에 따라 4가지 유형으로 분류하였으며, 자극성 섬유종과 10예의 정상 구강점막에 톨루이딘 블루 염색과 CD 68 면역조직화학염색을 시행하였다. 이를 통계화하여 자극성 섬유종의 조직학적 아형에 따른 비만 세포와 대식 세포의 분포 정도를 관찰하였다. 연구결과 : 통계 결과 비만 세포와 대식 세포의 분포는 자극성 섬유종에서 현저히 증가하였으며, Spearman 상관계수는 0.693이었다. 결론 : 조직의 섬유화에 관여하는 비만 세포는 자극성 섬유종의 cellular type에서 유의하게 증가하였으며, 대식 세포도 자극성 섬유종의 모든 아형에서 유의하게 증가하였다. 따라서 자극성 섬유종의 형성 과정에는 비만 세포와 대식 세포의 증가가 중요한 역할을 하는 것으로 생각되었다.

• Journal of the Korean Dietetic Association
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• v.12 no.1
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• pp.82-88
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• 2006

This study was performed to evaluate the effect of Sorghum bicolor L. Moench(Sorghum, su-su) extracts on mouse immune cell activation. As ex vivo experiment, different concentrations(0, 50, 500mg/kg B.W.) of Sorghum bicolor L. Moench water extracts were orally administrated into mouse every other day for four weeks. The proliferation of mouse splenocytes, the number of plaque forming cells(PFC) and the cytokine IL-1β production by activated macrophage were used as indices for immunocompetence. Splenocyte proliferation was enhanced in mouse orally administrated with 50mg/kg B.W./day concentration compared to that of control group. Especially, the highest proliferation of spleoncyte was seen in the mouse orally administrated at the concentration of 50mg/kg B.W./day. The number of plaque forming cells(PFC) to SRBC were significantly enhanced when compared with control group. Also, the mouse of Sorghum bicolor L. Moench water extracts 50mg/kg B.W./day supplementation group with LPS stimulation enhanced level of IL-1$\beta$ cytokine production. This study suggest that supplementation of Sorghum bicolor L. Moench water extracts may enhance the immune function by regulating the splenocytes proliferation, increasing the number of PFC and enhancing the cytokine production by activated macrophage.

• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.1-10
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• 2005

Objective: We examined the antimetastatic effect of Curcuma zedoaria Roscoe (CZ) on pulmonary metastasis of B 16 cells. Methods: For 6 weeks, Zedoariae Rhizoma made from dried CZ were dissolved in distilled water and administered to mice 2 weeks before they were injected with B]6 melanoma cells. Mice were given CZ at doses of 250 and 500 mg/kg, and were compared for lung weight, survival days, and NO production. Results: Intake of CZ throughout the experiment extended the average survival time. Intake after B16 cell injection slightly prolonged survival time, but intake before B]6 cell injection did not influence life span. We examined the effect of CZ on macrophage function by measuring NO production. After the macrophages were given CZ for 6 weeks, the amount of NO generated by the macrophages stimulated with LPS in culture medium increased. NO generated by the macrophages also served as a cytotoxic factor against B16 melanoma cells. B16 melanoma-conditioned medium reduced NO production by macrophages. However, CZ treatment reversed the reduction in NO production by the conditioned medium significantly. Conclusion : These findings may suggest that macrophage function-modulating activity by CZ appears to underlie its antimetastatic activity, which leads to a decrease in the number of lung metastatic surface nodules and the extension of life span.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.628-634
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• 2002

A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.

• YAKHAK HOEJI
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• v.47 no.4
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• pp.234-238
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• 2003

Linarin is a main compound from Chrysanthemum zawadskii var, latilobum. However, the biological mechanisms of these activities are unclear. Because of this wide diversity of effects, it is believed that they may be exerted through pluripotent effectors of linarin. In our previous screening study, the effects of linarin on the mouse macrophages cell line, RAW 264.7 cells, were investigated. It was found that linarin could stimulate macrophages activation by the production of tumor necrosis factor (TNF). The linarin (6.25∼12.5 $\mu\textrm{g}$/mι) inhibited the production of NO in LPS-activated RAW 264.7 cells and linarin became an useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO. However, linarin-treated total lymphocyte showed cytotoxicity in a dose dependent manner between 20 $\mu\textrm{g}$/mι and 40 $\mu\textrm{g}$/mι. In this study, linarin derivative (acetylated linarin) was synthesized in order to obtain less-cytotoxicity of linarin and evaluated for their in vitro cytotoxic activity aganist mouse total lymphocyte. There was no cytotoxic activity in a dose dependent manner (20∼40 $\mu\textrm{g}$/mι) of acetylated linarin whereas linarin showed. The production of NO, however, was not the case by this modified linarin. The cell morphological change was not significantly changed in response to acetylated linarin alone and these effects were potentiated by the addition of LPS. These results suggest that acetylated linarin may be developed to be a promising new drug candidate without cytotoxicity on the basis of its activity of macrophage activation.