• Title/Summary/Keyword: Inflammatory gene expression

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Survey of the Expression Pattern and Immuno Stimulatory Effect of DNA Vaccine Using β-Galactosidase Reporter System in Olive Flounder (Paralichthys olivaceus)

  • Lee Sang-Jun;Hong Suhee;An Kyong-Jin;Kim Young-Ok
    • Fisheries and Aquatic Sciences
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    • v.7 no.2
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    • pp.70-75
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    • 2004
  • The CMV promoter driven lacZ reporter gene (pcDNA-lacZ) was constructed and used for DNA immunization study. The expression of the lacZ gene was confirmed in vitro using RTG-2 cell line before using for in vivo study in olive flounder (Paralichthys olivaceus). In the dose response study, the maximum expression of the lacZ gene was found in the group injected with 5 ${\mu} g$ of the plasmid DNA. Kinetic study showed a significantly increased expression of $\beta-galactosidase$ gene at 7 days after injection. Effects of DNA vaccine on specific and nonspecific immune responses such as antibody and NO production were studied and the significant effect was found in olive flounder injected with 10 and 15 ${\mu} g$ DNA (sub optimal dose for lacZ gene expression). Two pro inflammatory cytokine genes, $IL-l\beta$ and $TNF-\alpha$, were also found to be up regulated in the muscle injected with the plasmid, suggesting an induction of local inflammatory response.

Inhibition of p65 Nuclear Translocation by Radicicol, Heat Shock Protein Inhibitor

  • Kim, Sang-Gyu;Jeon, Young-Jin;Lee, Seog-Ki
    • Toxicological Research
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    • v.21 no.4
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    • pp.285-290
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    • 2005
  • We demonstrate that radicicol, a macrocyclic antifungal antibiotic originally isolated from Monosporium bonorden, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of peritoneal macrophages and RAW 264.7 cells with radicicol inhibited LPS-stimulated nitric oxide production in a dose-related manner. Immunohistochemical staining of iNOS and RTPCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression in RAW 264.7 cells. Immunostaining of p65, EMSA, and reporter gene assay showed that radicicol inhibited $NF-\kappa/Rel$ nuclear translocation. DNA binding, and transcriptional activation, respectively. Collectively, these series of experiments indicate that radicicol inhibits iNOS gene expression by blocking $NF-\kappa/Rel$ nuclear translocation. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of radicicol on iNOS suggest that radicicol may represent a useful anti-inflammatory agent.

Expression Patterns of Host Inflammatory Cytokine Genes during Infestation with Haemaphysalis longicornis, a Zoonotic Vector, in Blood Sucking Periods

  • Islam, Mohammad Saiful;You, Myung-Jo
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.53-59
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    • 2018
  • Tick saliva is critically important for continuous attachment to the host, blood feeding for days, and transmission of tick-borne pathogens. To characterize the patterns of inflammatory cytokine gene expression during its attachment and blood sucking time, peripheral blood samples of rabbits infested with Haemaphysalis longicornis ticks were collected at different intervals. Blood histamine concentration was evaluated as well as gene encoding IFN-${\gamma}$, TNF-${\alpha}$, IL-2, IL-6, IL-4, and IL-10 were compared with non-infested rabbits. Blood histamine concentration of tick-infested rabbits during fast feeding time was significantly higher than that of non-infested rabbits. In both nymph and adult tick infested rabbits, expression of TNF-${\alpha}$ and IFN-${\gamma}$ genes were decreased significantly (P<0.05), while expression of IL-4, IL-6, and IL-10 were increased 1.3 to 7 folds in adult infested rabbits with the exception of IL-6 that was significantly (P<0.05) decreased in nymph infested rabbits. IL-2 was not expressed in either nymph or adult infestation. H. longicornis saliva is capable of modulate host responses through a complex correlation with histamine and Th1, Th2 mediated cytokines that suppress the inflammatory responses directed toward inflammatory mediators introduced into the host during tick feeding.

Antibacterial Activity and Anti-inflammatory Effect of Methanol Extracts of Saliva miltiorrhiza Against Oral Pathogenic Bacteria (단삼 메탄올 추출물의 구강 병원성 세균에 대한 항균 및 항염증효과)

  • Lee, JungHyeok;Yim, Dongsool;Choi, SungSook
    • Korean Journal of Pharmacognosy
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    • v.52 no.1
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    • pp.41-48
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    • 2021
  • This research was conducted to investigate the antibacterial and anti-inflammatory effects of MeOH Ex. of Salvia miltiorrhiza (MESM) against oral pathogenic bacteria. Minimum inhibitory concentration (MIC), removal effect of biofilm produced by Streptococcus mutans, effect of gene expression of proinflammatory cytokines and effect of production of proinflammatory cytokine of MESM were tested. MESM showed moderated antibacterial activity against oral pathogenic bacteria. About 89±8% of biofilms produced by S. mutans were removed by MESM at a concentration of 1 mg/mL. Gene expression of IL-1β and TNF-α induced by Porphyromonas gingivalis were 8~9 folds reduced by MESM. Gene expression of IL-8 induced by Fusobacterium nucelatum were 12 folds reduced by MESM. Production of IL-1β, TNF-α and IL-8 were significantly suppressed by MESM. Conclusively, MESM showed potent antibacterial and anti-inflammatory effect against oral pathogenic bacteria.

Expression Profile of Inflammatory Genes in Human Airway Epithelial A549 Cells

  • Sohn, Sung-Hwa;Ko, Eun-Jung;Kim, Sung-Hoon;Kim, Yang-Seok;Shin, Min-Kyu;Hong, Moo-Chang;Bae, Hyun-Su
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.44-50
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    • 2009
  • This study was conducted to evaluate the inflammation mechanisms of tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-4 (IL-4), and IL-$1{\beta}$-induced stimulation of A549 human epithelial cells. In the present study, A549 cells were stimulated with TNF-$\alpha$, IL-4 and IL-$1{\beta}$ to induce expression of chemokines and adhesion molecules involved in eosinophil chemotaxis. The effects of TNF-$\alpha$, IL-4 and IL-$1{\beta}$ on gene expression profiles in A549 cells were evaluated by oligonucleotide microarray and Real time RT-PCR. The gene expression profiles for the A549 cells varied depending on the cytokines. Also, the results of the microarray and Real time RT-PCR revealed that inflammatory-related genes were up-regulated in cytokine stimulated A549 cells. Cytokines can affect inflammation in A549 cells. A microarray-based genomic survey is a high-throughput approach that enables evaluation of gene expression in cytokine stimulated cell lines.

Gold Sodium Thiomalate Inhibits iNOS Gene Expression in RAW 264.7 Macrophage: Differential Regulation by Gold Sodium Thiomalate and Sodium Salicylate (Gold Sodium Thiomalate에 의한 유도성 Nitric Oxide Synthase Gene의 발현억제: Gold 제제와 Sodium Salicylate의 차이점)

  • 임종호;배진영
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.230-235
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    • 2002
  • Gold sodium thiomalate (GST, gold compound) is a widely used anti-arthritic, anti-rheumatic and anti-inflammatory drug that is considered a good alternative to sodium salicylate (NaSA) for individuals who cannot tolerate salicylates. Nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation. Recent evidence suggests that anti-inflammatory effect of NaSA lies in the inhibition of iNOS, but nothing has been reported about the direct effect of iNOS expression by GST. The present study was designed to elucidate sequentially the action mechanisms of GST and NaSA on lipopolysaccharide (LPS) plus interferon-gamma (IFN-$\gamma$) induced iNOS expression in RAW 264.7 macrophages. Both GST and NaSA inhibited NO production and iNOS protein expression in a dose dependent manner. GST inhibited iNOS mRNA expression induced by LPS plus IFN-$\gamma$, whereas NaSA did not. These findings suggest that GST may exert anti-arthritic, anti-rheumatic and anti-inflammatory effect by inhibiting iNOS expression induced by LPS plus IFN-$\gamma$ at transcriptional level, whereas NaSA exert its effect by inhibiting iNOS expression at the translational or posttranslational level.

Chemokine Gene Expression in Mice during Orientia tsutsugamushi Infection

  • Koh, Young-Sang
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.266-270
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    • 2003
  • Orientia tsutsugamushi, an obligate intracellular bacterium, is the causative agent of scrub typhus which is histopathologically characterized by inflammatory manifestations. To understand the pathogenesis of scrub typhus, chemokine gene expression in mice after infection with O. tsutsugamushi was investigated. The mRNAs that were upregulated included macrophage inflammatory proteins 1${\alpha}$/${\beta}$ (MIP-1${\alpha}$/${\beta}$), MIP-2, monocyte chemoattractant protein 1, RANTES (regulated upon activation, normal T-cell expressed and secreted), and gamma-interferon-inducible protein 10. Peak expression of these chemokines was observed six days after infection. These responses returned to or approached baseline preinfection levels by eight days after infection. Chemokine profiles in infected mice were well correlated with the kinetics of inflammatory cell infiltration. Thus, O. tsutsugamushi appears to be a strong inducer of chemokines which may significantly contribute to the inflammation observed in scrub typhus by attracting and activating phagocytic leukocytes.

Effects of Nodakenin, Columbianadin, and Umbelliferone Isolated from the Roots of Angelica decursiva on the Gene Expression and Production of MUC5AC Mucin from Human Airway Epithelial NCI-H292 Cells

  • Lee, Hyun Jae;Lee, Choong Jae
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.201-207
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    • 2017
  • Angelica decursiva has been utilised as remedy for controlling the airway inflammatory diseases in folk medicine. We investigated whether nodakenin, columbianadin, and umbelliferone isolated from the roots of Angelica decursiva inhibit the gene expression and production of MUC5AC mucin from human airway epithelial cells. Confluent NCI-H292 cells were pretreated with nodakenin, columbianadin or umbelliferone for 30 min and then stimulated with epidermal growth factor (EGF), phorbol 12-myristate 13-acetate (PMA) or tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) for 24 h. The MUC5AC mucin gene expression was measured by reverse transcription - polymerase chain reaction (RT-PCR). Production of MUC5AC mucin protein was measured by enzyme-linked immunosorbent assay (ELISA). The results were as follows: (1) Nodakenin did not affect the expression of MUC5AC mucin gene induced by EGF, PMA or $TNF-{\alpha}$. Columbianadin inhibited the expression of MUC5AC mucin gene induced by EGF or PMA. However, umbelliferone inhibited the expression of MUC5AC mucin gene induced by EGF, PMA or $TNF-{\alpha}$; (2) Nodakenin also did not affect the production of MUC5AC mucin protein induced by EGF, PMA or $TNF-{\alpha}$. Columbianadin inhibited the production of MUC5AC mucin protein induced by PMA. However, umbelliferone inhibited the production of MUC5AC mucin protein induced by EGF, PMA or $TNF-{\alpha}$. These results suggest that, among the three compounds investigated, umbelliferone only inhibits the gene expression and production of MUC5AC mucin stimulated by various inducers, by directly acting on airway epithelial cells, and the results might explain the traditional use of Angelica decursiva as remedy for diverse inflammatory pulmonary diseases.

Healing Effect of Yukmijihwang-tang on Fracture Factor and Morphological Changes in Femur Fractured Mice (육미지황탕(六味地黃湯)이 대퇴골절 동물모델의 골절 유합인자 및 형태학적 변화에 미치는 영향)

  • Kim, Hyun-Seok;Jeon, Dong Hwi;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.4
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    • pp.17-30
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    • 2020
  • Objectives The purpose of this study is to evaluate the healing effect of Yukmijihwang-tang (YM) on femur fractured mice. Methods Mice were randomly divided into 6 groups: normal, control, positive control, YM with low, medium, high dosage each. All groups were prepared with femur fracture and treated diffrently. In order to measure bone regeneration effects, we analysed the levels of cyclooxygenase-2 (COX2), bone morphogenetic protein-2 (BMP2), collagen type II alpha 1 chain (Col2a1), Sox9, runt-related transcription factor 2 (Runx2), and osterix genes expressed in bone. For morphological analysis, muscles were removed and femur was observed with naked eye. Results COX2 gene expression in bone marrow significantly decreased. BMP2 gene expression significantly increased. Col2a1 gene expression significantly increased. Sox9 gene expression increased as well. Runx2 gene expression in bone marrow increased, but there was no statistical significance. Osterix gene expression significantly increased. Union of the fracture site progressed more in YM group compared to the control group. The fracture union score was significantly decreased in YM group compared to the control group. Conclusions YM showed anti-inflammatory effect, promoted bone regeneration by stimulating the bone regeneration factor. In conclusion, YM can help fracture healing and it well be applied clinically to patients with fracture.

Notochordal Cells Influence Gene Expression of Inflammatory Mediators of Annulus Fibrosus Cells in Proinflammatory Cytokines Stimulation

  • Moon, Hong-Joo;Joe, Hoon;Kwon, Taek-Hyun;Choi, Hye-Kyoung;Park, Youn-Kwan;Kim, Joo-Han
    • Journal of Korean Neurosurgical Society
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    • v.48 no.1
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    • pp.1-7
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    • 2010
  • Objective : Notochordal cells in the intervertebral disc interact with nucleus pulposus (NP) cells and support the maintenance of disc homeostasis by regulation of matrix production. However, the influence of notochordal cells has not been evaluated in the annulus fibrosus (AF), which is the primary pain generator in the disc. We hypothesized that the notochordal cell has the capacity to modulate inflammatory mediators secreted by AF cells secondary to stimulation. Methods : Notochordal and AF cells were isolated from adult New Zealand white rabbits. AF pellets were cultured with notochordal cell clusters or in notochordal cell-conditioned media (NCCM) for 24 or 48 hours with proinflammatory cytokines at varying concentrations. Gene expression in AF pellets were assayed for nitric oxide synthase (iNOS), cyclo-oxygenase (COX)-2, and interleukin (IL)-6 by real time reverse transcriptase polymerase chain reaction (RT-PCR). Results : AF pellet in NCCM significantly decreased the iNOS and COX-2 messenger ribonucleic acid (mRNA) levels compared to AF pellets alone and AF pellets with notochordal cells (p < 0.05). AF pellet resulted in dose-dependent iNOS and COX-2 expression in response to IL-$1{\beta}$, stimulation, demonstrating that 1 ng/ml for 24 hours yielded a maximal response. AF pellet in NCCM significantly decreased the expression of iNOS and COX-2 in response to 1ng/ml IL-$1{\beta}$, stimulation at 24 hours (p < 0.05). There was no difference in IL-6 expression compared to AF pellets alone or AF pellets with notochordal cell clusters. Conclusion : We conclude that soluble factors from notochordal cells mitigate the gene expression of inflammatory mediators in stimulated AF, as expected after annular injury, suggesting that notochordal cells could serve as a novel therapeutic approach in symptomatic disc development.