• 대한한방내과학회지
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• 제29권4호
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• pp.1048-1060
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• 2008

Cirsii Japonici Herba(CJ) is a wild perennial herb found in many areas of Korea as well as China and Japan, which has been used to treat bleeding and inflammation. Silibinin is the main flavonoid extracted from milk thistle (Cirsii Japonici Herba). It exhibits potent antioxidant activity and anti-inflammatory effect. In this study, the effect of CJ and silibinin extract on lipopolysaccharide-induced inflammation was investigated using MTS assay, RT-PCR, western blot, and nitric oxide detection on mouse BV2 microglial cell lines. In the present results, CJ and silibinin extract suppressed nitric oxide production by inhibiting the lipopolysaccharide-stimulated enhancement of COX-2 and iNOS gene expression in BV2 cells. Moreover, CJ and silibinin also repressed some lipopolysaccharide-induced signaling molecules. Importantly, catalase-induced COX-2 and iNOS expression needed activations of $NF-{\kappa}B$, PI3K/Akt, and MAPK, which were important for the transcriptional up-regulation of COX-2 and iNOS. CJ and silibinin interaction on BV2 cells down-regulated $NF-{\kappa}B$-dependent proinflammatory cytokine (IL-2,IL-6) expression. They are involved in the regulation of inflammatory responses. These data shows that CJ and silibinin exerts anti-inflammatory and analgesic effects, probably by suppression of COX-2 and iNOS synthase expression in BV2 microglial cells.

• 대한의생명과학회지
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• 제19권2호
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• pp.118-123
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• 2013

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a proinflammatory cytokine that mediates the inflammatory response and immune functions, and modulates the proliferation, differentiation and cell death of cancer cells. The differential functions of TNF-${\alpha}$ in various human cells due to the formation of different stimulating pathway upon the binding of TNF-${\alpha}$ to its receptors. In the present study, we examined the different effects of TNF-${\alpha}$ on the survival and apoptosis between normal granulocytes and human myeloid leukemia HL-60 cells. Although TNF-${\alpha}$ did not affect on the constitutive apoptosis of granulocytes, TNF-${\alpha}$ strongly induced the apoptosis of HL-60 cells in a dose- and a time-dependent manner. TNF-${\alpha}$-induced apoptosis was occurred via the activation of caspase 8, caspase 9 and caspase 3/7 and the induction of ROS production in HL-60 cells. Also, BAY-11-7085, a NF-${\kappa}B$ inhibitor, blocked the TNF-${\alpha}$-induced apoptosis in HL-60 cells. NF-${\kappa}B$ may be involved in TNF-${\alpha}$-induced apoptotic signaling pathway in HL-60 cells. These results suggest that TNF-${\alpha}$ activates apoptotic pathways and its process depends on cell type and many cellular factors. A better understanding of the differential effect of TNF-${\alpha}$ on cell apoptosis and survival may provide important information that can be used to elucidate the specific inhibitory effect of TNF-${\alpha}$ on the cancer dis.

• The Korean Journal of Physiology and Pharmacology
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• 제19권2호
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• pp.183-189
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• 2015

Foeniculum vulgare Mill. (fennel) is used to flavor food, in cosmetics, as an antioxidant, and to treat microbial, diabetic and common inflammation. No study to date, however, has assessed the anti-inflammatory effects of fennel in experimental models of inflammation. The aims of this study were to investigate the anti-inflammatory effects of fennel in model of lipopolysaccharide (LPS)-induced acute lung injury. Mice were randomly assigned to seven groups (n=7~10). In five groups, the mice were intraperitoneally injected with 1% Tween 80-saline (vehicle), fennel (125, 250, $500{\mu}l/kg$), or dexamethasone (1 mg/kg), followed 1 h later by intratracheal instillation of LPS (1.5 mg/kg). In two groups, the mice were intraperitoneally injected with vehicle or fennel ($250{\mu}l/kg$), followed 1 h later by intratracheal instillation of sterile saline. Mice were sacrificed 4 h later, and bronchoalveolar lavage fluid (BALF) and lung tissues were obtained. Fennel significantly and dose-dependently reduced LDH activity and immune cell numbers in LPS treated mice. In addition fennel effectively suppressed the LPS-induced increases in the production of the inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, with $500{\mu}l/kg$ fennel showing maximal reduction. Fennel also significantly and dose-dependently reduced the activity of the proinflammatory mediator matrix metalloproteinase 9 and the immune modulator nitric oxide (NO). Assessments of the involvement of the MAPK signaling pathway showed that fennel significantly decreased the LPS-induced phosphorylation of ERK. Fennel effectively blocked the inflammatory processes induced by LPS, by regulating pro-inflammatory cytokine production, transcription factors, and NO.

• Journal of Microbiology and Biotechnology
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• 제30권3호
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• pp.333-340
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• 2020

Macrophages are the cells of the first-line defense system, which protect the body from foreign invaders such as bacteria. However, Gram-negative bacteria have always been the major challenge for macrophages due to the presence of lipopolysaccharides on their outer cell membrane. In the present study, we evaluated the effect of phloretin, a flavonoid commonly found in apple, on the protection of macrophages from Escherichia coli infection. RAW 264.7 cells infected with standard E. coli, or virulent E. coli K1 strain were treated with phloretin in a dose-dependent manner to examine its efficacy in protection of macrophages. Our results revealed that phloretin treatment reduced the production of nitric oxide (NO) and generation of reactive oxygen species along with reducing the secretion of proinflammatory cytokines induced by the E. coli and E. coli K1 strains in a concentration-dependent manner. Additionally, treatment of phloretin downregulated the expression of E. coli-induced major inflammatory markers i.e. cyclooxygenase-2 (COX-2) and hemeoxygenase-1 (HO-1), in a concentration dependent manner. Moreover, the TLR4-mediated NF-κB pathway was activated in E. coli-infected macrophages but was potentially downregulated by phloretin at the transcriptional and translational levels. Collectively, our data suggest that phloretin treatment protects macrophages from infection of virulent E. coli K1 strain by downregulating the TLR4-mediated signaling pathway and inhibiting NO and cytokine production, eventually protecting macrophages from E. coli-induced inflammation.

• IMMUNE NETWORK
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• 제3권1호
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• pp.61-68
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• 2003

Background: Fibroblast functions both as a structural element and as a vital immunoregulatory cell. Fibroblasts regulate inflammation through governing of chemokine expression. In order to elucidate the mechanisms by which the expressions of chemokines were regulated, the co-stimulatory effects of Th1 and proinflammatory cytokines were compared using nasal mucosal fibroblasts. Methods: Human nasal mucosa was obtained from surgery for septal deviation and the growth of fibroblasts was established. Fibroblasts from 4th to 6th passage were stimulated with various combinations of cytokines. To inhibit selected signaling pathways, fibroblasts were pretreated with cyclosporin A, wortmannin, staurosporine, and dexamethasone prior to the stimulation with cytokines. The supernatants were collected and chemokines were detected with a sandwich enzyme-linked immunosorbent assay. Results: $TNF-{\alpha}/IFN-{\gamma}$-induced production of RANTES was inhibited by all inhibitors used. MCP-1 was produced constitutively and $TNF-{\alpha}$-induced or $TNF-{\alpha}/IFN-{\gamma}$-induced production of MCP-1 was not inhibited by cyclosporin A or wortmannin, but by stauroporine or dexamethasone. All inhibitors used in this experiment inhibited $TNF-{\alpha}/IFN-{\gamma}$-induced or $IL-1{\beta}/IFN-{\gamma}$-induced production of MCP-2 in nasal mucosal fibroblasts. Although staurosporine or dexamethasone showed strong inhibitory effects, cyclosporin A or wortmannin did not inhibit the production of MCP-3 by $IL-1{\beta}/IFN-{\gamma}$ treatment. Conclusion: Chemokines were strongly induced by stimulation of cytokines in combination and showed different pattern of inhibition by the inhibitors. Therefore, it was assumed that cytokines acted on multiple pathways or on unknown pathways which converged to gene-specific transcription factors.

• Journal of Acupuncture Research
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• 제24권3호
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• pp.197-205
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• 2007

Objectives : Tumor necrosis factor-${\alpha}$(TNF ${\alpha}$) is a proinflammatory cytokine involved in the pathogenesis of rheumatoid arthritis. This study was designed to investigate the relation between TNF-${\alpha}$ gene polymorphism and rheumatoid arthritis in Korean population. Methods : This study was carried out on 103 rheumatoid arthritis patients who fulfilled the American College of Rheumatology 1987 revised criteria for rheumatoid arthritis and 208 healthy control subjects. Blood samples from all subjects were obtained for DNA extraction. The extracted DNA was amplified by polymerse chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. We investigated the genotyping of TNF-${\alpha}$ by using Pyrosequencing. Results: The genotypes of TNF-${\alpha}$ gene were GG, AG and AA. While the distribution of TNF-${\alpha}$ polymorphism in control subjects was 92.31%, 7.21%, 0.48% respectively, in rheumatoid arthritis patients was 93.20%, 6.80%, 0.00%(GG, AG, AA). There was no statistical significant allelic frequency difference between control and rheumatoid arthritis groups. Conclusions : We concluded that there was no significant association between TNF-${\alpha}$ gene polymorphism and rheumatoid arthritis. However, the findings of this study need to be confirmed in more patients and further studies.

• 대한본초학회지
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• 제26권2호
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• pp.75-81
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• 2011

Objectives : This study aims at examining the anti-inflammatory effects of Scutellariae Radix extract. Methods : Scutellariae Radix was hot water extracted to make the samples(SR) for the experiment. Their effects were examined on the increase of cell viability in mouse macrophage Raw 264.7 cells, the creation of nitric oxide(NO) in lipopolysaccharide(LPS)-induced Raw 264.7 cells, and the creation of cytokines of interleukin(IL)-$1{\beta}$ and others. Results : The results of the experiment are as follows. 1. The MTT assay was carried out to check the cellular toxicity of the water extract of Scutellariae Radix. The results were found no significant toxicity caused to macrophages by the water extract of Scutellariae Radix. 2. The water extract of Scutellariae Radix significantly restricted the increase of NO in the LPS-induced macrophages after 24-hour culture. 3. The water extract of Scutellariae Radix significantly restricted the creation of IL-6, IL-10, IL-12p40, IL-17, interferon-inducible protein(IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor(VEGF) in the LPS-induced macrophages at the concentration of $25{\mu}g/mL$ or higher. Conclusion : The samples(SR) of hot water extract of Scutellariae Radix caused no significant cellular toxicity to macrophages and significantly restricted the creation of NO, IL-6, IL-10, IL-12p40, IL-17, IP-10, KC, and VEGF in the LPS-induced macrophages at $25{\mu}g/mL$ or higher, thus demonstrating significant anti-inflammatory effects.

• BMB Reports
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• 제33권5호
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• pp.402-406
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• 2000

In order to understand the role of AGP on the differentiation of promyelocytic leukemia cells, the AGP expression and its relation to cytokines were investigated during granulocytic or monocytic differentiation of HL-60 cells. When HL-60 cells were treated with all-trans-retinoic acid (ATRA) for 5 days, the cells were fully differentiated into granulocytes, and the AGP mRNA and protein levels were continuously increased up to 5 days in a dose- and time- dependent manner. However, in the case of the monocytic differentiation of HL-60 cells by tetradeanoyl phorbol acetate (TPA), the AGP gene expression was not induced. In addition, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNAs were also enhanced during granulocytic differentiation. These cytokine transcripts showed a peak level 3 days after the ATRA treatment. It decreased gradually thereafter. However, direct addition of recombinant cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) and dexamethasone to the HL-60 cell cultures showed no AGP induction. These findings suggest that the AGP and proinflammatory cytokines are expressed in ATRA-treated promyelocytic cells. However, these cytokines do not act as autocrine inducers on AGP expression. This fact implies that the AGP expression during granulocytic differentiation of HL-60 cells is induced through a signal pathway different from hepatocyte signaling in inflammation.

• 동의신경정신과학회지
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• 제19권3호
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• pp.85-100
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• 2008

Objective: This experiment was designed to investigate the effect of the KSKH hot water extract & ultra-fine powder on microglia and memory deficit model. Method: The effects of the KSKH hot water extract on expression of IL-1$\beta$, IL-6, TNF-$\alpha$ mRNA and production of IL-1$\beta$, IL-6, TNF-$\alpha$ in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. The effects of the KSKH hot water extract & ultra-fine-fine powder on the behavior of the memory deficit mice induced by scopolamine and AChE in serum of the memory deficit mice induced by scopolamine were investigated. Results: 1. The KSKH hot water extract suppressed the expression of IL-1$\beta$, IL-6, TNF-$\alpha$ mRNA in BV2 microglial cell line treated by LPS. 2. The KSKH hot water extract suppressed the production of IL-1$\beta$, IL-6, TNF-$\alpha$ in 100$\mu g/m\ell$ concentration of BV2 microglial cell line culture supernatant. 3. The KSKH hot water extract & ultra-fine powder decreased AChE activation significantly in the serum of the memory deficit mice induced by scopolamine. 4. The KSKH hot water extract & ultra-fine powder showed significant effect on memory impairment in the stop-through latency type of Morris water maze test. Conclusions: This experiment shows that the KSKH hot water extract & ultra-fine powder might be effective for the prevention and treatment of amnesia and Alzheimer's disease. Investigation into the clinical use of the KSKH hot water extract & ultra-fine powder for amnesia and Alzheimer's disease is suggested for future research.

• Biomolecules & Therapeutics
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• 제5권2호
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• pp.165-173
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• 1997

This study was conducted to investigate the effect of DA-9601, an extract of Artemisiae Herba, which is known to possess mucoprotective action either by free radical scavenging effect or increase of mucus secretion, against animal models of inflammatory bowel disease (IBD) induced by trinirobenzene sulfonic acid (TNBS) or other noxious agents. Experimental colitis was induced by intracolonic administration of TNBS in 50% ethanol, or 1 ml of 7% acetic acid solution (AA), by subcutaneous injection of indomethacin (INDO) in rats, or by supplementing drinking water with 5% dextran sodium sulfate (DSS) in albino mice. DA-9601 was treated orally for 4 to 7 days. Animals were euthanized 1 day after the last treatment for morphological and biochemical analysises. All the noxious agents including TNBS, AA, INDO and DSS elicited severe colitis. The animals treated with DA-9601 showed a consistent, dose-related reduction in the severity of colitis, grossly and histologically. The reduction was significant (p<0.05) after administration of DA-9601 at dose range of 10 mg/kg or above. In TNBS-induced colitis, the rats receiving DA-9601 showed significantly decreased mucosal myeloperoxidase (MPO) and thiobarbituric acid-reactive substances (TBA-RS), when compared to control and mesalazine groups. Mucosal proinflammatory cytokine levels were also decreased after DA-9601 treatment. In conclusion, DA-9601 ameliorated macroscopic and histologic scores in experimental colitis either through decreasing oxidative stress or by attenuating cytokines involved in inflammation. DA-9601 could be a promising drug for the therapy of IBD.