• International Journal of Oral Biology
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• 제35권2호
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• pp.43-49
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• 2010

Enterococcus faecalis, a gram-positive bacterium, has been implicated in endodontic infections, particularly in chronic apical periodontitis. Proinflammatory cytokines, including tumor necrosis factor-$\alpha$ (TNF-$\alpha$), are involved in the pathogenesis of these apical lesions. E. faecalis has been reported to stimulate macrophages to produce TNF-$\alpha$. The present study investigated the mechanisms involved in TNF-$\alpha$ production by a murine macrophage cell line, RAW 264.7 in response to exposure to E. faecalis. Both live and heat-killed E. faecalis induced high levels of gene expression and protein release of TNF-$\alpha$. Treatment of RAW 264.7 cells with cytochalasin D, an inhibitor of endocytosis, prevented the mRNA up-regulation of TNF-$\alpha$ by E. faecalis. In addition, antioxidant treatment reduced TNF-$\alpha$ production to baseline levels. Inhibition of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinase also significantly attenuated E. faecalis-induced TNF-$\alpha$ expression by RAW 264.7 cells. Furthermore, activation of NF-${\kappa}B$ and AP-1 in RAW 264.7 cells was also stimulated by E. faecalis. These results suggest that the phagocytic uptake of bacteria is necessary for the induction of TNF-$\alpha$ in E. faecalis-stimulated macrophages, and that the underlying intracellular signaling pathways involve reactive oxygen species, ERK, p38 MAP kinase, NF-${\kappa}B$, and AP-1.

• Neonatal Medicine
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• 제18권1호
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• pp.42-48
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• 2011

목적: 기관지폐이형성증(bronchopulmonary dysplasia,BPD)의 발생에는 폐의 염증반응이 중요하게 작용한다. Tumor necrosis factor-$\alpha$ (TNF-$\alpha$)는 대표적인 proinflammatory cytokine으로, TNF-$\alpha$ 고생산 부위의 단일염기다형성과 BPD발생 사이의 연관성을 알아보고자 하였다. 방법: 2006년 1월부터 12월까지 서울대학교병원에서 출생하여 신생아중환자실에 입원한 재태주령 32주 미만이면서 출생체중 1,500 g 미만의 미숙아 93예를 대상으로, 실시간 중합효소연쇄반응을 이용하여, TNF-$\alpha$ 유전자 촉진자 5개 부위(-1031T/C,-863C/A, -857C/T, -308G/A, -238G/A)의 단일염기다형성을 분석하였다. 결과: BPD 군은 전체연구대상93예 중에서 38예로40.9%에 해당하였다. BPD 군에서 대조군보다 재태연령($27^{+5}{\pm}2^{+0}$ wk vs. $29^{+2}{\pm}1^{+4}$ wk, P<0.0001)과 출생체중(990${\pm}$270 g vs. 1,220${\pm}$230 g, P<0.0001)이 낮고, 신생아 호흡곤란증후군(71.1% vs. 49.1%, P=0.035) 과 동맥관개존(71.1% vs. 50.9%, P=0.052) 의 발생빈도가 높아서 두 군간의 임상적 특성에는 차이가 있었다. 그러나 TNF-$\alpha$ 유전자 단일염기다형성 분석에서는, 각각의 TNF-$\alpha$ 대립유전자의 빈도와 일배체형의 분포 모두가 BPD 군과 대조군 사이에 차이를 보이지 않았다. 결론: 한국인 조산아에서는, TNF-$\alpha$ 유전자 다형성보다는 임상적인 위험인자가 BPD 발생에 중요하게 작용하였다.

• Journal of Genetic Medicine
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• 제6권2호
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• pp.155-160
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• 2009

목 적: 자간전증은 인간의 임신 특이적 증후군으로 임신 기간 동안 감소된 자궁 관류 압에 의해 나타나는 태반 허열에 의해 시작된다. 자간전증은 염증성 싸이토카인의 비정상적인 발현과 연관되어 있는 것으로 알려져 있다. 싸이토카인 중 대표적인 종양 사멸 인자-알파(tumor necrosis factor-alpha; TNF-alpha)는 자간전증 여성에서 증가되는 것으로 보고되었다. 하지만 TNF-alpha 유전자 다형성과 자건전증 사이의 연관성에 관한 연구는 미비한 실정이다. 따라서 이번 연구에서는 TNF-alpha 유전자 프로모터 지역의 C-850T의 단일염기다형성을 한국인 자간전증 여성에서 확인하고 자간전증의 발달과의 연관성을 연구하고자 한다. 대상 및 방법: 이 유전자 다형성은 SNapShot kit와 ABI Prism3100 Genetic analyzer를 사용하여 198명의 자간전증 임산부와 194명의 정상 임산부의 말초 혈액에서 분석하였다. 결 과: C-850T 유전자형과 대립유전자 빈도는 자간전증 임산부와 정상 임산부 사이에 차이가 없었다. 유전자형인CC, CT, TT는 자간전증 임산부에서 각각 74.3%, 22.2%, 3.5% 였고, 정상 임산부에서 71.6%, 25.8%, 2.6%였다. 그리고 C와 T 대립유전자 빈도는 자간전증 임산부에서 각각 0.85, 0.15 였고 정상 임산부에서 0.84, 0.16였다. 자간전증 발생 위험도는 C-850T의 이종접합 유전자형(CT)이나 돌연변이 유전자형(TT)을 수반하는 그룹에서 증가되지 않았다. 결 론: 우리는 이번 연구에서 자간전증과 정상 임신부 사이에 C-850T의 유전자형과 대립유전자 빈도는 차이가 없음을 발견했다. 따라서 이번 연구는 TNF-alpha 유전자 다형성인 C-850T가 한국인 임신부의 자간전증 발생과 관련이 없을 가능성을 시사한다.

• 대한한방내과학회지
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• 제25권1호
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• pp.28-45
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• 2004

Objectives : The main purpose of this study is to evaluate the effect of Injinchunggan-tang on $TNF-{\alpha}$ signal transmission system. Materials and Methods : We analyzed the following with quantitative RT-PCR method; the effect of Injinchunggan-tang on secretion of $TNF-\alpha$ mRNA/protein and stability, the effect on gene revelation that consists of signal transmission system (TRAIL, NIK, A20, TRADD, RAIDD, RIP TNFR-I, TNFR-II, TRAF1, TRAF2, FADD), the one on activation of p38, Erk1/2 MAPK and the rate of nuclear $NF-{\kappa}B/cytosolic\;NF-{\kappa}B$ in HepG2 cell. We also analyzed the inhibitory effect of Injinchunggan-tang on the apoptosis of HepG2 cell that $TNF-{\alpha}$ induces and the $NF-{\kappa}B$ restraint effected by transfection of $I{\kappa}B{\Delta}N$ through tryphan blue exclusion assay. Results : Injinchunggan-tang prohibits revelation of $TNF-{\alpha}$ mRNA in HepG2 cell and the creation of protein. However, it has no effect on the stability of $TNF-{\alpha}$ mRNA. While it did not have any effect on the generation of TRAIL, NIK, A20, TRADD, RAIDD and RIP genes, Injinchunggan-tang reduces the revelation of TNFR-I, TNFR-II, TRAF1, TRAF2 and FADD genes. It has been confirmed that Injinchunggan-tang restraints the revelation of $TNF-{\alpha}$ mRNA that is promoted by ethanol, acetaldehyde, lipopolysaccharide, in proportion to the treatment density and time. It activated $NF-{\kappa}B$ of HepG2 cell and promoted activation of $NF-{\kappa}B$ that is occurred by $TNF-{\alpha}$. It has been observed that the restraint effect against the $TNF-{\alpha}$ inducing apoptosis is lost when it is intercepted the function of $NF-{\kappa}B$ in HepG2 cell. Conclusion: It has been confirmed that Injinchunggan-tang has restraining effect against the revelation of $TNF-{\alpha}$ and mRNA that is constituent element of TNF-a signal transmission system. It also has been revealed that it restraints the activation of p38, Erk1/2 by $TNF-{\alpha}$. Through this prohibiting effect, it is inferred that it restraints signal transmission among various cells that are related to inflammation reaction. Meanwhile, Injinchunggan-tang protects liver cell from apoptosis that is caused by $TNF-{\alpha}$, by maintaining the activating function for $NF-{\kappa}B$.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6217-6220
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• 2012

Introduction: Up to present, EGF $61^*A$/G, TGF-${\beta}1$-$509^*T$/C and TNF-${\alpha}$-$308^*A$/G gene polymorphisms have been analysed in other cancer entities than hepatocellular carcinoma (HCC). We here investigated the frequency of these gene polymorphisms among HCC patients. Materials and Methods: A total of 73 HCC patients and 117 cancer-free healthy people were recruited at the Surgical Department of Zhongshan Hospital. Genomic DNA was isolated from peripheral blood and gene polymorphisms were analyzed by PCR-RFLP. Results: The distribution of EGF $61^*G$/G homozygotes among HCC patients was more frequent than that in the control group (24.7% vs 11.1%, OR=2.618, 95%CI=1.195-5.738). In parallel, the frequency of the "G" allele in the HCC patient group was also higher than that in the control group (45.9% vs 33.3%, OR= 1.696, 95%CI=1.110-2.592). No difference could be found for the TGF-${\beta}1$-509 and TNF-${\alpha}$-308 genotypes. Conclusion: EGF $61^*G$/G genotype and G allele are significantly increased among patients with HCC. TGF-${\beta}1$-$509^*T$/C and TNF-${\alpha}$-$308^*A$/G gene polymorphisms are not related to this cancer entity.

• 약학회지
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• 제56권3호
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• pp.198-203
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• 2012

Chrysin and chlorogenic acid are natural products derived from Scutellariae Radix and Lonicerae Flos, respectively. We examined whether chrysin and chlorogenic acid affect airway mucin production induced by TNF-${\alpha}$ in NCI-H292 cells. Cells were pretreated with each agent for 30 min and then stimulated with TNF-${\alpha}$ for 24 h. Of the two compounds, chrysin suppressed airway MUC5AC mucin production. Also, chrysin suppressed MUC5AC mucin gene expression and translocation of NF-${\kappa}B$ p65 induced by TNF-${\alpha}$. This result suggests that chrysin can regulate the production and gene expression of mucin induced by TNF-${\alpha}$ through the inactivation of NF-${\kappa}B$ in airway epithelial cells.

• 대한한방소아과학회지
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• 제28권2호
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• pp.56-71
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• 2014

Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

• 한방안이비인후피부과학회지
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• 제23권2호
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• pp.57-68
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• 2010

Objective : This study was performed to evaluate the effect of Lithospermum erythrorhizon extracts on the inflammatory cytokines gene expression by the bacteria of Propionibacterium acnes (P. acnes) which elicits acne in human monocytes, THP-1 cell line. Experiment : Cytotoxicity of Lithospermum erythrorhizon extracts was analyzed by XTT assay. Real time RT-PCR was applied to analyze the cytokines gene expressions of IL-8, MCP-1 and TNF-$\alpha$. Translocation of transcription factor NF-${\kappa}B$ from cytoplasm into nucleus was observed using immunocytochemistry and confocal microscopy. Results : Lithospermum erythrorhizon extracts did not show cytotoxicity as high as in $1,000\;{\mu}g/ml$ of concentration. Transcription levels of inflammatory cytokines, IL-8, MCP-1 and TNF-$\alpha$ were increased by P. acnes in THP-1 and Lithospermum erythrorhizon extracts decreased the upregulated transcription levels. Lithospermum erythrorhizon extracts significantly inhibited the translocation of NF-${\kappa}B$ into nucleus by P. acnes. Conclusion : This study suggests that Lithospermum erythrorhizon extracts have anti-inflammatory effects on P. acnes treated THP-1 as decreasing the mRNA expressions of IL-8, MCP-1 and TNF-$\alpha$. This anti-inflammatory effect of Lithospermum erythrorhizon extracts may be useful in therapeutic treatments for acne vulgaris.

• 대한한의학회지
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• 제24권2호
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2004년도 International Meeting of the Microbiological Society of Korea
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• pp.65-67
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• 2004

O-GlcNAcylation of p53 has been already identified and reported, but the function of O-GlcNAc on p53 has not been studied well. In this report, the general function of O-GlcNAc modification on p53 has been investigated using mouse fibroblast cell, L929. When streptozotocin (STZ), a non-competitive O-GlcNAcase inhibitor was treated to L929, O-GlcNAc modification level was dramatically increased on nucleocytoplasmic proteins, including p53. Because it has been already reported that $TNF\alpha$ induced the production of p53 in L929, $TNF\alpha$ was treated to obtain more p53. Approximately two times more amount of p53 was found from the cells treated STZ and $TNF\alpha$ simultaneously compared to the cell treated $TNF\alpha$ alone. The p53 increment in the presence of STZ was not caused by the induction of p53 gene expression. When new production of p53 induced by the $TNF\alpha$ was inhibited by the treatment of cycloheximide, O-GlcNAc modification decreased and phosphorylation increased on pre-existing p53 after $TNF\alpha$ treatment. But in the presence of STZ and $TNF\alpha$ at the same time, more O-GlcNAcylation occurred on p53, The level of ubiquitination on p53 was also reduced in the presence of STZ. Approximately three times less amount of Mdm2 bound to this hyperglycosylated p53. From this result it might be concluded that treatment of STZ to inhibit O-GlcNAcase increased O-GlcNAc modification level on p53 and the increment of O-GlcNAc modification stabilized p53 from ubiquitin proteolysis system.