• Journal of Oral Medicine and Pain
• /
• 제24권4호
• /
• pp.361-374
• /
• 1999

F. nucleatum is a gram-negative obligate anaerobe which is the principal and most frequent cause of gingival inflammation and is the predominant pathogen isolated in subsequent periodontal breakdown. It is also one of the most numerous bacteria found in subgingival plaque samples from healthy sites; its numbers are about 10-fold greater in plaque from periodontally diseased sites. The purpose of this study is to examine the effects of outer membrane(OM), outer membrane vesicle(OMV), and lipopolysaccharide(LPS) from F. nucleatum ATCC 25586 strain on the growth of human gingival fibroblasts and HOS 941 cells, and on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes. For the examination of cytotoxic effects, $TNF-{\alpha}$ production and $TNF-{\alpha}$ mRNA expression, the MTT assay, the ELISA and the RT-PCR were performed, respectively. All extracts of F. nucleatum tested were cytotoxic to both of human gingival fibroblasts and HOS 941 cells, and the significant difference of cytotoxic activity among the extracts was not observed. In the effects of these extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes, all extracts of F. nucleatum tested also stimulated the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression, but the effects of the OM extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression were higher than those of the OMV and the LPS extracts. The pattern of the $TNF-{\alpha}$ mRNA expression was similar to that of the $TNF-{\alpha}$ production. These results indicate that F. nucleatum seems to contribute to the pathogenesis of periodontal diseases at least by its cytotoxicity, directly and its $TNF-{\alpha}$ production, indirectly.

• Biomolecules & Therapeutics
• /
• 제6권1호
• /
• pp.31-36
• /
• 1998

Capsular polysaccharides (CPs) from Streeptococcus pneumoniae were examined for the ability to induce secretory responses in a pure population of peritoneal macrophages. The highly purified CPs were able to affect the macrophage, ie, secretion of tumor necrosis factor-alpha (TNF-$\alpha$) and nitrite. As after stimulation with CPs, secretion of TNF-u induced by these CPs reached its maximum within the first few hours of the interaction, while secretion of nitrite was increased with time. In addition, production of TNF-$\alpha$ and nitrite was increased in a dose-dependent manner. In the presence of indomethacin, CP-stimulated TNF-$\alpha$ production was not altered. In contrast, LPS with indomethacin stimulated 24.5% more TNF-$\alpha$ than LPS alone, suggesting that the intracellular signaling processes for TNF production are differentially stimulated by CP and LPS. The results demonstrate that CPs are potent inducer of macrophage secretory activities.

• 한국축산식품학회지
• /
• 제21권4호
• /
• pp.374-382
• /
• 2001

Lf는 cytokines의 생성 및 면역반응 등의 생체 방어적 기능을 하는 물질로 알려져 있다. 본 연구에서는 Lf과 Lf-h에 의해 macrophage에서 TNF-$\alpha$와 NO 생성에 미치는 효과를 조사하였다. K-Lf 및 K-Lf-h는 단독으로 TNF-$\alpha$의 생성을 증가시켰으며, Lf의 농도에 따라 그 생성량이 증가하였다. LPS와 함께 작용시켰을 경우에는 큰 효과가 없는 것으로 나타났으나, 세포 성장률은 증가시켰다. 그러나 B-Lf, H-Lf, 그리고 이들의 가수분해물들은 단독으로는, RAW264.7 cell을 자극하여 TNF-$\alpha$나 NO의 생성을 증가시키지 못하였다. 또한, K-Lf는 그 자체만으로 TNF-$\alpha$에서 보여준 것처럼 NO생성에 영향을 미치며 농도가 높아짐에 따라 NO 생성을 증가시키는 경향을 보여주었다.

• 약학회지
• /
• 제42권1호
• /
• pp.82-88
• /
• 1998

The effect of hepatoprotective agents and bile acids on tumor necrosis factor-alpha, (TNF-${\alpha}$) production in murine and human macrophage cell line (RAW264.7 and U937) was inve stigated. The hepatoprotective agents including silymarin and its major component, silybin, significantly inhibited TNF-alpha production in a concentration dependent manner ($IC_50$ of silybin=67.7${\mu}g$/ml (140.3${\mu}g$M)). In differentiated U937 cells, especially, silybin showed more effective inbitory activity ($IC_50$=35.1${\mu}g$g/ml (72.7${\mu}g$M)). These results suggest that silymarin and silybin may inhibit TNF-alpha production in the process of hepatic diseases in human. However, biphenyldimethyl dicarboxylate (DDB) was not effective. In the case of bile acids, chenodeoxycholic acid (CDCA) showed a concentration dependent inhibitory effect on TNF-alpha production ($IC_50$ of CDCA= 71.5${\mu}g$g/ml (182.1${\mu}g$M)). In contrast, glycine or taurine conjugated form (G-CDCA or T-CDCA) restored to the control level or significantly increased TNF-${\alpha}$ production. And also ursodeoxycholic acid (UDCA) and its conjugated forms (G-UDCA and T-UDCA) showed a variety of patterns on TNF-${\alpha}$ production by changes of functional groups and concentration. These results also indicate that bile acids may regulate TNF-${\alpha}$ production in normal hepatic function or disease conditions.

• Journal of Periodontal and Implant Science
• /
• 제40권3호
• /
• pp.119-124
• /
• 2010

Purpose: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-$\alpha$ production in differentiated THP-1 cells, a human monocytic cell line. Methods: LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-$\alpha$ and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-$\alpha$ and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. Results: Leptin enhanced P. intermedia LPS-induced TNF-$\alpha$ production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-$\alpha$ expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-$\alpha$ production was not mediated by the leptin receptor. Conclusions: The ability of leptin to enhance P. intermedia LPS-induced TNF-$\alpha$ production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

• The Korean Journal of Physiology
• /
• 제30권1호
• /
• pp.77-83
• /
• 1996

Platelet-activating factor(PAF) is a phospholipid mediator of pulmonary inflammation, and immunologic reaction. In this study, the role of PAF on tumor necrosis factor$(TNF_{-{\alpha}})$ production by rat alveolar macrophages(AM) was examined. When PAF $(10^{-12}{\sim}10{-16}\;M)$ alone was added to AM culture, $(TNF_{-{\alpha}})$ production was not significantly increased above the resting level. In contrast, the combined addition of PAF $(10^{-6}\;M)$ and muramyl dipeptide(MDP) $(1.0\;{\mu}g\ml)$ to AM cultures markedly enhanced $(TNF_{-{\alpha}})$ production with 8.2 fold increase compared with AM culture in resting state. This potentiative effect was 313% above the sum of the separate effects of PAF and MDP. To characterize MDP effects on $(TNF_{-{\alpha}})$ production, the dose-response of AM cultured with various concentrations of MDP was tested. High level of MDP $(10\;{\mu}g\ml)$ could not significantly enhance the potentiation effect on $(TNF_{-{\alpha}})$ production compared with AM cultures with low level of MDP $(0.1\;{\mu}g\ml)$, i.e. 112.5% vs 107.8%, respectively when $10^{-10}$ M of PAF was simultaneously added to the cell culture. These data support that the potentiation of TNF. g production in AM culture is mediated by PAF rather than MDf It was also evaluated whether the similar result was obtained in silica, respirable toxic particle-treated AM culture. $(TNF_{-{\alpha}})$ production was also significantly enhanced in the PAF $(10^{-6}\;M)$ and silica $(50\;{\mu}g\ml)$-added cell cultures with 4.7 fold above the value of silica alone-stimulated cells. These results indicate that PAF can potentiate $(TNF_{-{\alpha}})$ production by MDP-or silica- stimulated AM and suggest that PAF may play a potent role in lung inflammation and disease associated with microbe and occupational dust exposures.

• 동의신경정신과학회지
• /
• 제11권2호
• /
• pp.11-21
• /
• 2000

Substance P(SP) can stimulate production of tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) from astrocytes stimulated with lipopolysaccharide(LPS). The objective of the current study was to determine the effect of Taraxacum officinale(TO) on the production of TNF-${\alpha}$ from primary cultures of rat astrocytes. TO(100& 1000$\mu\textrm{g}$/ml) significantly inhibited the TNF-${\alpha}$ production by astrocytes stimulated with LPS and SP. Interleukin-1(IL-1) has been shown to elevate TNF-${\alpha}$ production from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore examined whether IL-1 mediated inhibition of TNF-${\alpha}$ production from primary astrocytes by TO. Treatment of TO(100 and 1000$\mu\textrm{g}$/ml) to astrocytes stimulated with both LPS and substance P decreased IL-1 production significantly. Moreover, the production of TNF-${\alpha}$ by LPS and substance P in astrocytes was progressively inhibited with increasing amount of IL-1 neutralizing antibody. These results suggest that TO may inhibit TNF-${\alpha}$ production by inhibiting IL-1 production and that TO has an antiinflammatory activity in the central nervous system.

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

• The Korean Journal of Physiology and Pharmacology
• /
• 제16권2호
• /
• pp.131-138
• /
• 2012

Alcoholic hepatitis is a leading cause of liver failure in which the increased production of tumor necrosis factor ${\alpha}$ (TNF${\alpha}$) plays a critical role in progression of alcoholic liver disease. In the present study, we investigated the effects of cilostazol, a selective inhibitor of type III phosphodiesterase on ethanol-mediated TNF${\alpha}$ production in vitro and $in$ $vivo$, and the effect of cilostazol was compared with that of pentoxifylline, which is currently used in clinical trial. RAW264.7 murine macrophages were pretreated with ethanol in the presence or absence of cilostazol then, stimulated with lipopolysacchride (LPS). Cilostazol significantly suppressed the level of LPS-stimulated TNF${\alpha}$ mRNA and protein with a similar degree to that by pentoxifylline. Cilostazol increased the basal AMP- activated protein kinase (AMPK) activity as well as normalized the decreased AMPK by LPS. AICAR, an AMPK activator and db-cAMP also significantly decreased TNF${\alpha}$ production in RAW264.7 cells, but cilostazol did not affect the levels of intracellular cAMP and reactive oxygen species (ROS) production. The $in$ $vivo$ effect of cilostazol was examined using ethanol binge drinking (6 g/kg) mice model. TNF${\alpha}$ mRNA and protein decreased in liver from ethanol gavaged mice compared to that from control mice. Pretreatment of mice with cilostazol or pentoxifylline further reduced the TNF${\alpha}$ production in liver. These results demonstrated that cilostazol effectively decrease the ethanol-mediated TNF${\alpha}$ production both in murine macrophage and in liver from binge drinking mice and AMPK may be responsible for the inhibition of TNF${\alpha}$ production by cilostazol.

• 한국임상수의학회지
• /
• 제31권6호
• /
• pp.469-476
• /
• 2014

본 연구는 염증상태에서의 CLA의 효과와 작용기전을 알아보기 위해 LPS-자극 RAW 264.7 macrophages에 있어 ROS 생성과 TNF-${\alpha}$ 생산, NF-${\kappa}B$ 및 $PPAR{\gamma}$ 활성을 검토하였다. t10c12-CLA는 LPS로 자극하지 않은 비염증시의 RAW 세포에서는 ROS 생성을 증가시켜 TNF-${\alpha}$ 생산을 유도하였으며, 이 효과는 $PPAR{\gamma}$ 활성화에 의존해서 NF-${\kappa}B$ 활성 증가에 의해 매개되었다. 반면, LPS로 자극한 염증조건의 RAW 세포에서는 t10c12-CLA가 $PPAR{\gamma}$ 활성화에 의존하지 않는 경로로 ROS 생성 및 과도한 TNF-${\alpha}$ 생산을 억제하였다. 본 결과로부터 CLA는 ROS 생성을 통해 TNF-${\alpha}$ 생산 및 NF-${\kappa}B$ 활성을 염증 유무에 따라 조절하는 것으로 사료되었다.