• Journal of Pharmacopuncture
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• v.6 no.3
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• pp.39-47
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• 2003

Background & Methods : In order to measure the acute and subacute toxicity of Vermilionum and it's anti-cancer effects, Sarcoma-180 abdominal cancer cells were injected intravenously. The following results were obtained after measuring the survival rate, toxicity of the NK cells, and IL- 2 productivity. Results : 1. It was impossible to measure $LD_{50}$ value in the acute toxicity test and no toxic effects were witnessed in the clinical observation. 2. No significant differences were shown in the weight changes between the experiment groups and the control group in the acute toxicity test. 3. No peculiar toxic effects were shown in the subacute toxicity test and the weight changes were insignificant between the experiment groups and the control group. 4. In measuring the survival rate after inducing abdominal cancer by Sarcoma-180, the experiment groups showed increased of 9.52% compared to the control group. 5. In measuring the activity of NK cells, no significant changes were shown between the experiment groups and the control group. 6. In measuring the productivity of IL-2, significant reduction was shown in the experiment groups compared to the normal group, but no significance was witnessed compared to the control group.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.46-46
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• 2021

The first purpose of this study was to evaluate the scavenging capacity (SC) of DPPH and ABTS free radicals for ethanol extract (STR-E) and its active fractions from Sophora tonkinensis root (STR). Four different fractions from STR-E were prepared by using different types of solvents such as chloroform (STR-E-C), ethyl acetate (STR-E-EA), n-butanol (STR-E-B), and water (STR-E-W). STR-E-C showed the highest value of total phenolic content, while STR-E showed the highest value of total flavonoid and terpenoid content. In STR-E and its four fractions, STR-E-EA showed the strongest SC with the lowest SC50 values of the DPPH radicals and ABTS radicals. The second purpose of this study was to evaluate anti-inflammatory activity in the lipopolysaccharide (LPS)-induced RAW 264.7 macrophages treated with STR-E, STR-E-C, and STR-E-EA, respectively. No cytotoxic effect to RAW 264.7 cells was observed at 20 ~ 25 ㎍/ml of STR-E, 10 ㎍/ml of STR-E-C, and 5 ㎍/ml of the STR-E-EA, presenting cell viability values close to that of the untreated control (100%). STR-E, STR-E-C, and STR-E-EA significantly suppressed the LPS-induced nitric oxide (NO) in a dose-dependent manner. Results of reverse-transcription (RT)-qPCR analysis showed that the peak mRNA levels of IL-1β, TNF-α, iNOS, IL-6, and IL-10 were observed in the LPS-stimulated macrophages at 4 h, 2 h, 12 h, 12 h, and 12 h, respectively. The peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced in the LPS-stimulated macrophages co-treated with 20 ㎍/ml and 25 ㎍/ml of STR-E, respectively. In the case of IL-10, its peak mRNA level slightly increased without statistical significance. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 10 ㎍/ml and 20 ㎍/ml of STR-E-C, respectively. In contrast, the peak mRNA level of IL-10 significantly increased at 8 h. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 5 ㎍/ml and 10 ㎍/ml of STR-E-EA, respectively. In contrast, the peak mRNA level of IL-10 increased at 4 h. Taken together, our data indicated that STR-E, STR-E-C, and STR-E-EA activate macrophages to secrete both pro-inflammatory and anti-inflammatory cytokines.

• Korean Journal of Plant Resources
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• v.34 no.5
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• pp.411-419
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• 2021

In this study, we investigated in vitro immune-enhancing and anti-obesity activity of Abelmoschus manihot roots (AMR) in mouse macrophage RAW264.7 cells and mouse adipocytes 3T3-L1 cells. AMR increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in RAW264.7 cells. The inhibition of toll like receptor (TLR) 2 and 4 blocked AMR-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of mitogen-activated protein kinases (MAPKs) signaling pathway reduced AMR-mediated production of immunostimulatory factors. From these results, AMR is considered to have immune-enhancing activity through TLR2/4-mediated activation of MAPKs signaling pathway. In addition, AMR inhibited lipid accumulation and reduced the protein level such as CCAAT enhancer-binding protein alpha (CEBPα), peroxisome proliferator-activated receptor gamma (PPARγ), perilipin-1, adiponectin and fatty acid binding protein 4 (FABP4) associated with lipid accumulation in 3T3-L1 cells, indicating that AMR may have anti-obesity activity. Based on these results, AMR is expected to be used as a potential functional agent for immune enhancement and anti-obesity.

• IMMUNE NETWORK
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• v.5 no.1
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• pp.11-15
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• 2005

Background: Throughtout the last three decades, the therapy of leukemias and lymphoma has set the stage for curative cancer therapy in systemic malignant disease. This was the result of an integrated work of basic reaserch and clinical investigators leading to more aggressive albeit tolerable protocol of chemotherapy and radiotherapy. High dose therapy marks the most elaborated strategies in this field today. However, intensification of conventional therapeutic modalities as mentioned has to be based on new approaches and the exploration of new antineoplastic mechanisms. This insight has resulted in immune therapy of cancer. Among the cells of the immune system, natural killer (NK) cells and T cells are of major interest for the development of therapeutic strategies. Methods: Cytotoxicity to target cells was measured by LDH release method, Characterization of activated lymphocyte was measured by Flow cytometry analysis. Anti-CD3, 16, 56 monoclonal antibody and IL-2 were used for the activation of NK and T cell. The analysis of effect of activated lymphocyte, in vivo, were used by Balb/c nude mouse. Results and Conclusion: Cytotoxicity to K562 cells was significantly higher in the mixture group of NK and T cells than that of a group of activating T cells. The survivors and the rate of reduction of size of tumor craft of nude mouse group treatment with activated lymphocyte was higher than that of the group without treatment with activated lymphocyte. Therefore, this results are suggested that the activated lymphocytes by anti-CD3, CD16 and CD56 can reduce the malignancy effect of lymphoma.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.23-28
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• 2015

Objectives : The aim of this study is to investigate cell viability, anti-inflammatory, antioxidant, immunoenhancing activity using various extracts of Yeonsan Ogye.Methods : In order to evaluate cytotoxicity, MTT assay was performed. We investigated production levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-αand interleukin (IL)-6, and nitric oxide(NO) in LPS-induced RAW 264.7 cells. NO production in RAW 264.7 cells was measured by using Griess reagent. Cytokines including IL-6 and TNF-αwere measured by Luminex and ROS was measured by Flow cytometry.Results : No cytotoxicity of various extracts of Yeonsan Ogye was observed in RAW 264.7 cells. Productions of ROS in RAW 264.7 cells were increased from extraction of bones and decreased from extraction of skin. Also, productions of NO in RAW 264.7 cells were increased to bone extract and decreased at skin extract. In addition, productions of pro-inflammatory cytokines (IL-6 and TNF-α) in LPS-induced RAW 264.7 cells were decreased at skin, meat extracts, respectively. Finally, the levels of immune-related cytokines (IL-6 and TNF-α) were increased compared to those of the normal group.Conclusions : It is concluded that Yeonsan Ogye extracts seem to have significant biological activities likes anti-inflammatory, antioxidant, immuno-enhancing etc. These results may be developed as a raw material for new health food and new therapeutics to ease the symptoms related with inflammatory and oxidative stress. In terms of oriental traditional medicine, we expect that it contribute to building of EBM (Evidence-Based Medicine) from the this result.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.785-791
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• 2005

Corydalis Tuber has traditionally been used for the treatment of water retention in the body. Administration of the aqueous extract of Corydalis Tuber has been known to be effective for the control of pain and treatment of arthritis. It was reported that Corydalis Tuber possesses anti-inflammatory activity and modulates the intestinal immune system. The effect of Corydalis Tuber against LPS-stimulated expressions of COX-2, iNOS, and $IL-1{\beta}$ in cells of the murine RAW 264.7 macrophages was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), $PGE_2$ immunoassay, and NO detection. The aqueous extract of Corydalis Tuber was shown to suppress $PGE_2$ production by inhibition on the LPS-stimulated enhancement of COX-2 enzyme activity, $IL-1{\beta}$, and iNOS expression in the RAW 264.7 macrophages. Present results suggest that Corydalis Tuber exerts anti-inflammatory and analgesic effects probably by suppressing of COX-2, iNOS, and $IL-1{\beta}$ expressions, resulting in inhibition of $PGE_2$ synthesis. Corydalis Tuber has anti-inflammatory and analgesic effects probably by suppressing of COX-2, iNOS, and $IL-1{\beta}$ mRNA expressions, resulting in inhibition of $PGE_2$ and NO synthesis.

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.79-84
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• 2016

Objective : Nypa fruticans Wurmb. Fruit (NF) has been used as a conventional medicine to treat inflammatory peridontal diseases in Myanmar and Eastern Asia. However, the anti-inflammatory effect of NF aqueous extract on lipopolysaccharide (LPS)-induced inflammatory responses was not well-investigated. Therefore, this study was aimed to investigate the anti-inflammatory effect of NF on LPS-induced inflammatory responses on RAW 264.7 cells.Methods : To induce inflammation on the macrophage cell line, RAW 264.7 cells were treated with 500 ng/mL of LPS. Water extracts of NF was treated 1 h prior to treatment of LPS. Cell viability was measured by MTT assay. Production of nitrite was measured with Griess assay and pro-inflammatory cytokines such as interleukine (IL)-1β and IL-6, and tumor necrosis factor (TNF)-α was measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR). In addition, we examined the inhibitory mechanisms of NF by western blot and immunocytochemistry.Result : Water Extract from NF itself did not have any cytotoxic effect at the concentration of 200 ㎍/ml in RAW 264.7 cells. Treatment of NF inhibited the production of nitrite, and pro-inflammatory cytokines inlcuding IL-1β, IL-6 and TNF-α in a dose dependant. In addition, NF treatment inhibited the LPS-induced activation and translocation of nuclear factor (NF)-κB.Conclusion : In summary, our result suggest that treatment of NF could reduce the LPS-induced inflammatory responses via deactivation of NF-κB. This study could suggest that NF could be a beneficial drug or agent to prevent inflammation.

• Journal of Acupuncture Research
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• v.36 no.4
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• pp.256-263
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• 2019

Background: The purpose of this study was to investigate the anti-inflammatory response of lipopolysaccharide (LPS) activated macrophages (RAW 264.7 murine cell line) to JCE003 which is an extract including Eucommia ulmoides, Juglans regia, Eleutherococcus senticosus, and Zingiber officinale. Methods: An MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed to analyze the survival rate of RAW 264.7 cells. The production of nitric oxide and pro-inflammatory cytokines (IFN-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in LPS-induced RAW 264.7 cells was measured by enzyme-linked immunosorbent assay. mRNA expression levels of pro-inflammatory cytokines (IFN-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6) were analyzed by quantitative polymerase chain reaction analysis. Results: Exposure of LPS-activated RAW 264.7 cells to JCE003 was not cytotoxic up to $400{\mu}g/mL$, but cell survival was statistically significantly decreased at $800{\mu}g/mL$ (p < 0.001). Nitric oxide production was not markedly lowered in LPS-activated RAW 264.7 cells by exposure to JCE003 (10, 50, 100, 200, 400, $800{\mu}l/mL$) compared with the Control group. In addition, JCE003 reduced the production of TNF-${\alpha}$ in LPS-induced RAW 264.7 cells at $400{\mu}g/mL$ (p < 0.05), but IFN-${\gamma}$ and TNF-${\alpha}$ mRNA expression in LPS-induced RAW 264.7 cells was decreased at 100, 200, and $400{\mu}g/mL$ JCE003 (p < 0.01). Conclusion: These results suggest that JCE003 inhibited the expression and production of pro-inflammatory cytokines in LPS-activated RAW 264.7 cells. The findings of this study provide basic data for the development of new Korean medicine anti-inflammatory drugs.

• Journal of Radiation Industry
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• v.5 no.3
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• pp.273-277
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• 2011

This study evaluated the cytotoxicity and anti-inflammatory activities of natural herbal extracts including Houttuynia cordata and Eucommia ulmoides against human mast cell (HMC-1). Houttuynia cordata (HC) and Eucommia ulmoides(EU) were extracted with distilled water (at $75^{\circ}C$) and then freeze-dried for 5 days. Finally, the mixture of HC and EU were sterilized by ${\gamma}$-rays irradiation. Cytotoxicity of the mixture against HMC-1 cell was measured using cell counting kit-8 (CCK-8) assay. In addition, inflammatory mediator cytokines such as TNF-${\alpha}$, IL-6 and IL-8 were evaluated by ELISA kit on the HMC-1 cells with calcimycin A23187 and phorbol 12-myristate 13-acetate (PMA). The results showed that mixture of HC and EU had no cytoxicity and reduced TNF-${\alpha}$, IL-6, IL-8 response on HMC-1 cells.

• Journal of Periodontal and Implant Science
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• v.42 no.6
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• pp.185-195
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• 2012

Purpose: (-)-epigallocatechin-3-gallate (EGCG) has been reported to exert anti-inflammatory and antibacterial effects in periodontitis. However, its exact mechanism of action has yet to be determined. The present in vitro study evaluated the anti-in-flammatory effects of EGCG on human periodontal ligament fibroblasts (hPDLFs) and human periodontal ligament stem cells (hPDLSCs) affected by bacterial lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis. Methods: hPDLFs and hPDLSCs were extracted from healthy young adults and were treated with EGCG and/or P. gingivalis LPS. After 1, 3, 5, and 7 days from treatment, cytotoxic and proliferative effects were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine assay, respectively. And then, the gene expressions of hPDLFs and hPDLSCs were observed for interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor (TNF)-${\alpha}$, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL), and RANKL/OPG using real-time polymerase chain reaction (PCR) at 0, 6, 24, and 48 hours after treatment. The experiments were performed with the following groups for hPDLFs and hPDLSCs; 1) No treat, 2) EGCG alone, 3) P. gingivalis LPS alone, 4) EGCG+P. gingivalis LPS. Results: The 20 ${\mu}M$ of EGCG and 20 ${\mu}g/mL$ of P. gingivalis LPS had the lowest cytotoxic effects, so those concentrations were used for further experiments. The proliferations of hPDLFs and hPDLSCs increased in all groups, though the 'EGCG alone' showed less increase. In real-time PCR, the hPDLFs and hPDLSCs of 'EGCG alone' showed similar gene expressions to those cells of 'no treat'. The gene expressions of 'P. gingivalis LPS alone' in both hPDLFs and hPDLSCs were highly increased at 6 hours for IL-$1{\beta}$, IL-6, TNF-${\alpha}$, RANKL, and RANKL/OPG, except the RANKL/OPG in hPDLSCs. However, those increased gene expressions were down-regulated in 'EGCG+P. gingivalis LPS' by the additional treatment of EGCG. Conclusions: Our results demonstrate that EGCG could exert an anti-inflammatory effect in hPDLFs and hPDLSCs against a major pathogen of periodontitis, P. gingivalis LPS.