• Journal of Life Science
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• v.5 no.1
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• pp.20-25
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• 1995

기생충감영시 cytokine으로 활성화된 대식세포가 방어기전의 Effector cell로 작용할 때 분비하는 nitric oxide의 양 및 TNF-$\alpha$ 와 IFN-$\gamma$의 분비정도와 nitric oxide와의 상관관계 등을 알아보기 위하여 3종의 흡충류, Fasciola, paragonimus, Schistosma의 조항원 (100mg/ml)을 마우스 복강내에 주사　 후 24시간, 72시간, 9일간격으로 마우스의 대식세포(1X10$^{6}$/ml)를 분리하여 RPMI 배지 (10% FCS 첨가)에서 48시간 배양후 Nitric TNF-$\alpha$ 및 IFN-$\gamma$를 ELISA로 정량하여 다음과 같은 결과를 얻었다. Nitrite 생성정도는 Fasciola 조항원으로 24시간 감작시킨 대식세포에서 가장 높게 나타났으며 (140$\mu$M/ml) Paragonimus 항원군에서는 24시간에 최고치에 달하였다가(34 $\mu$M/ml) 시간이 경과함에 따라 점차 감소하였다. IFN-$\gamma$는 Paragonimus 항원군에서만 대조군에 비해 높았으며 9일경에 최고치를 보였다(475ng/ml). TNF-$\alpha$는 Schistosoma 항원군에서는 nitric oxide의 생성과 분비 양상이 일치하였다. 위의 결과에 의하면, 흡충류항원으로 감작된 마우스 대식세포의 nitric oxide 생성에 영향을 미치는 cytokine의 종류는 흥충류에 따라 차이를 보였으며, 이 중 Paragonimus 항원에 의해서는 IFN-$\gamma$의 분비가 촉진되는 것으로 나타났고, Schistosoma 의 경우에는 TNF-$\alpha$가 nitric oxide의 생성에 관계함을 알 수 있었다.

• Clinical and Experimental Pediatrics
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• v.46 no.7
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• pp.702-709
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• 2003

Purpose : $IFN{\gamma}$ sentitizes many tumor cells to $TNF{\alpha}$ and FASL-mediated apoptosis by enhancing the expression of TNF or FAS/CD95 receptor and modulating the activation of caspase and Bcl-2 family. It has been reported that $IFN{\gamma}$ and $TNF{\alpha}$ synergistically caused differentiation and growth inhibition of neuroblastoma cells. Even though some neuroblastoma cell express FASR/FASL on the cell surface, they could not induce apoptosis by ligation of the FAS/CD95 receptor. But the treatment of $IFN{\gamma}$ is reported to induce apoptosis in some neuroblastoma cell lines through the CD95/CD95L autocrine circuit. In this study, we examined whether $IFN{\gamma}$ could affect $TNF{\alpha}$ and agonistic FAS/CD95 antibody(CH-11)-induced apoptosis against neuroblastoma cell lines that had shown diverse drug sensitivity and resistance. Methods : CHLA-15, CHLA-90 and LA-N-2 neuroblastoma cells were cultured in IMDM and treated with recombinant $IFN{\gamma}$, $TNF{\alpha}$ and CH-11 antibody. Cell viability was measured by DIMSCAN with a fluorescent calcein-AM. Apoptosis was analyzed through flow cytometry using Annexin V-PE and 7-ADD staining and confirmed by pancaspase and caspase-8 blocking experiments. The expression of TNF RI and FAS/CD95 receptor was evaluated by flow cytometry using the corresponding antibody and PE-conjugated secondary antibody. Results : $IFN{\gamma}$ or $TNF{\alpha}$ alone had no demonstrable cytotoxic effects, whereas both cytokines in combination induced apoptosis synergistically in CHLA-15 and CHLA-90 cells. Although there was no cytotoxicity with the ligation of CH-11 alone in CHLA-90 cells, pretreatment of $IFN{\gamma}$ increased the sensitivity of CH-11-mediated apoptosis. The expression of TNFRI and FAS/CD95R were non-specifically enhanced after treatment of $IFN{\gamma}$ without relation to sensitivity to $TNF{\alpha}$ and CH-11. This finding suggest up-regulation of both receptors may contribute to sensitization of $TNF{\alpha}$ and CH-11-mediated apoptosis by $IFN{\gamma}$ in only sensitive cell lines. Conclusion : $IFN{\gamma}$ induced sensitization of $TNF{\alpha}$ and agonistic FAS/CD95 antibody-mediated apoptosis on some neuroblastoma cells through up-regulation of TNFRI and FAS/CD95 receptor.

• Herbal Formula Science
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• v.12 no.1
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• pp.209-223
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• 2004

Objective: This study was conducted to investigate the effects of Coptidis Rhizoma on the plasma IL-6 and $TNF-{\alpha}$ level in mice by intracerebroventricular(I.C.V.) injection of Lipopolysaccharide (LPS). Method: 6 mice were assigned to each of the Normal group, the Control group, and the individual Experimental groups. In the Normal group only saline was administered intragastrically, and in the Control group LPS was injected intracerebroventricularly 1 hr after intragastric administration of saline. In the Experimental groups Coptidis Rhizoma(0.5g/kg, 1.0g/kg, 3.0g/kg) was administered intragastrically to mice 1 hr prior to LPS (100ng/mouse) I.C.V. Injection. To measure the plasma IL-6 and $TNF-{\alpha}$ level of mice, their blood samples were collected from retro-orbital venous plexus, immediately centrifuged at $4^{\circ}C$, and plasma was removed and stored frozen at $-83^{\circ}C$ for later determination of plasma IL-6 and $TNF-{\alpha}$. Result: 1. LPS I.C.V. Injection increased plasma IL-6 level significantly in a dose-dependent manner compared with Normal group. (P<0.01) The plasma IL-6 concentration reached a significant maximal level about 1 hr after LPS(100ng/mouse) I.C.V. Injection.(P<0.001) 2. Both the 0.5g/kg(Sample A) and 1.0g/kg(Sample B) groups to which Coptidis Rhizoma was administered intragastrically 1 hr prior to LPS(100ng/mouse) I.C.V. Injection showed insignificant lower plama IL-6 level in 1 hr than Control group(P>0.05), and 3.0g/kg group(Sample C) conversely showed higher plama IL-6 level than Control group. 3. LPS I.C.V. Injection increased plasma $TNF-{\alpha}$ level significantly in a dose-dependent manner compared with Normal group.(P<0.05) The plasma $TNF-{\alpha}$ concentration reached a significant maximal level about 1 hr after LPS(100ng/mouse) I.C.V. Injection.(P<0.001) 4. All Sample groups(0.5g/kg, 1.0g/kg, and 3.0g/kg) to which Coptidis Rhizoma was administered intragastrically with each constituent-dose 1 hr prior to LPS(100ng/mouse) I.C.V. Injection showed significant lower $TNF-{\alpha}$ plama level in 1 hr than Control group.(P<0.001) These data revealed that Coptidis Rhizoma might have anti inflammatory effect by reducing the plasma $TNF-{\alpha}$ level in a dose dependent manner in mice LPS I.C.V. Injection.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.452-461
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• 1994

Background: Tumor necrosis factor(TNF)-$\alpha$ and Interleukin(lL)-$1{\beta}$ are thought to play a major role in the pathogenesis of the septic syndrome, which is frequently associated with adult respiratory distress syndrome(ARDS). In spite of many reports for the role of TNF-$\alpha$ in the pathogenesis of ARDS, including human studies, it has been reported that TNF-$\alpha$ is not sensitive and specific marker for impending ARDS. But there is a possibility that the results were affected by the diversity of pathogenetic mechanisms leading to the ARDS because of various underlying disorders of the study group in the previous reports. The purpose of the present study was to evaluate the roles of TNF-$\alpha$ and IL-$1{\beta}$ as a predictable marker for development of ARDS in the patients with septic syndrome, in which the pathogenesis is believed to be mainly cytokine-mediated. Methods: Thirty-six patients of the septic syndrome hospitalized in the intensive care units of the Asan Medical Center were studied. Sixteens suffered from ARDS, whereas the remaining 20 were at the risk of developing ARDS(acute hypoxemic respiratory failure, AHRF). In all patients venous blood samples were collected in heparin-coated tubes at the time of enrollment, at 24 and 72 h thereafter. TNF-$\alpha$ and IL-$1{\beta}$ was measured by an enzyme-linked immunosorbent assay (ELISA). All data are expressed as median with interquartile range. Results: 1) Plama TNF-$\alpha$ levels: Plasma TNF-$\beta$ levels were less than 10pg/mL, which is lowest detection value of the kit used in this study within the range of the $mean{\pm}2SD$, in all of the normal controls, 8 of 16 subjects of ARDS and in 8 in 20 subjects of AHRF. Plasma TNF-$\alpha$ levels from patients with ARDS were 10.26pg/mL(median; <10-16.99pg/mL, interquartile range) and not different from those of patients at AHRF(10.82, <10-20.38pg/mL). There was also no significant difference between pre-ARDS(<10, <10-15.32pg/mL) and ARDS(<10, <10-10.22pg/mL). TNF-$\alpha$ levels were significantly greater in the patients with shock than the patients without shock(12.53pg/mL vs. <10pg/mL) (p<0.01). There was no statistical significance between survivors(<10, <10-12.92pg/mL) and nonsurvivors(11.80, <10-20.8pg/mL) (P=0.28) in the plasma TNF-$\alpha$ levels. 2) Plasma IL-$1{\beta}$ levels: Plasma IL-$1{\beta}$ levels were less than 0.3ng/mL, which is the lowest detection value of the kit used in this study, in one of each patients group. There was no significant difference in IL-$1{\beta}$ levels of the ARDS(2.22, 1.37-8.01ng/mL) and of the AHRF(2.13, 0.83-5.29ng/mL). There was also no significant difference between pre-ARDS(2.53, <0.3-8.34ngfmL) and ARDS(5.35, 0.66-11.51ng/mL), and between patients with septic shock and patients without shock (2.51, 1.28-8.34 vs 1.46, 0.15-2.13ng/mL). Plasma IL-$1{\beta}$ levels were significantly different between survivors(1.37, 0.4-2.36ng/mL) and nonsurvivors(2.84, 1.46-8.34ng/mL). Conclusion: Plasma TNF-$\alpha$ and IL-$1{\beta}$ level are not a predictable marker for development of ARDS. But TNF-$\alpha$ is a marker for shock in septic syndrome. These result could not exclude a possibility of pathophysiologic roles of TNF-$\alpha$ and IL-$1{\beta}$ in acute lung injury because these cytokine could be locally produced and exert its effects within the lungs.

• Bulletin of the Korean Chemical Society
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• v.32 no.12
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• pp.4215-4220
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• 2011

This study introduces a new concept for a simple, efficient and cheap sensitivity amplification of a Quartz Crystal Microbalance (QCM) based immunosensor system for the detection of tumor necrosis factor-alpha (TNF-${\alpha}$, TNF) by using an in-built magnetic system. The frequency shift due to the applied magnetic field was successfully observed on magnetic particles labeled detection antibodies, anti-human TNF-${\alpha}$, which were bound to the immunologically captured TNF-${\alpha}$ on the gold coated quartz crystals. In the present system, the magnitude of frequency shift depends on both the strength of magnetic field and the amount of target antigen applied. Significant signal amplification was observed when the additional built-in residual stress generated by the modified magnetic particles under the magnetic field applied. Used in conjunction with a sandwich type non-competitive immunoassay format, the lower detection limit was calculated to be 25 $ngmL^{-1}$ and showed good linearity up to TNF-${\alpha}$ concentrations as high as 2.0 ${\mu}gmL^{-1}$. The sensitivity, most importantly, was improved up to 4.3 times compared with the same QCM system which was used only an antigen-antibody binding without additional magnetic amplification.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.1
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• pp.11-18
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• 1999

Nuclear factor $_{\kappa}B\;(NF-_{\kappa}B)$ activation is modulated by various protein kinases. Activation of $NF-_{\kappa}B$ is known to be important in the regulation of cell viability. The present study investigated the effect of inhibitors of protein tyrosine kinase (PTK), protein kinase C (PKC) and protein kinase A (PKA) on $NF-_{\kappa}B$ activity and the viability of bovine cerebral endothelial cells (BCECs). In serum-deprivation-induced BCEC death, low doses of $TNF{\alpha}$ showed a protective effect. $TNF{\alpha}$ induced $NF-_{\kappa}B$ activation within 4 h in serum-deprivation. PTK inhibitors (herbimycin A and genistein) and PKC inhibitor (calphostin C) prevented $NF-_{\kappa}B$ activation stimulated by $TNF{\alpha}.$ Likewise, these inhibitors prevented the protective effect of $TNF{\alpha}.$ In contrast to $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activity, basal $NF-_{\kappa}B$ activity of BCECs in media containing serum was suppressed only by calphostin C, but not by herbimycin A. As well BCEC death was also induced only by calphostin C in serum-condition. H 89, a PKA inhibitor, did not affect the basal and $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activities and the protective effect of $TNF{\alpha}$ on cell death. These data suggest that modulation of $NF-_{\kappa}B$ activation could be a possible mechanism for regulating cell viability by protein kinases in BCECs.

• Journal of Veterinary Clinics
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• v.28 no.2
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• pp.183-189
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• 2011

The objective of this study was to examine the effect of fucoidan on the phagocytic capapcity and oxidative burst activity (OBA) of canine peripheral blood polymorphonuclear cells (PMNs). The phagocytic capacity and OBA of PMNs were evaluated simultaneously by using a flow cytometer. Fucoidan itself did not cause any direct effect on the phagocytic capacity and OBA of PMNs. However, the phagocytic capacity and OBA of PMNs were enhanced by the culture supernatant from PBMCs treated with fucoidan. The phagocytic capacity and OBA of PMNs were also increased by treatment with recombinant canine (rc) tumor necrosis factor (TNF)-${\alpha}$. The ability of the culture supernatant from fucoidan-treated PBMCs to stimulate the phagocytic capacity and OBA of PMNs was inhibited by addition of anti-rc TNF-${\alpha}$ polyclonal antibody (PAb) prior to the culture. The amount of TNF-${\alpha}$ in the culture supematant from PBMCs was shown to increase upon treatment of fucoidan as compared with that of vehicle-treated PBMCs culture supematant. The level of TNF-${\alpha}$ mRNA expression in PBMCs was also up-regulated by the fucoidan treatment. These results suggest that fucoidan has an immunoenhancing effect on the phagocytic capacity and OBA of canine PMNs, which is mainly mediated by TNF-${\alpha}$ released from fucoidan-stimulated PBMCs.

• The Journal of Internal Korean Medicine
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• v.22 no.1
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• pp.79-85
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• 2001

Object : Hepatitis Treatment-tang No.1 has been used for the treatment of Liver disease and Jaundice. Long-term EtOH exposure leads to immunoregulatory and detoxification impairment. This study aimed to determine the relationship between TNF-${\alpha}$ production and expression, and EtOH-induced cytotoxicity on Hep G2 cells. Method : Cells were incubated with EtOH in the presence or absence of HT. The cells were tested after 24 hours and, again, after 48 hours. Cytoviability and TNF-${\alpha}$ release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability decreased, and the release of TNF-${\alpha}$ was increased. Increased amounts of TNF-${\alpha}$ contribute to EtOH-induced cytotoxicity. The Anti-TNF-${\alpha}$ antibody almost abolished it. Interestingly, EtOH-induced cytotoxicity and TNF-${\alpha}$ production were inhibited by HT. Moreover, when HT was used in combination with the anti-TNF-${\alpha}$ antibody, there was a marked inhibition of EtOH-induced cytotoxicity. Results : These results suggest that HT may prevent the cytotoxicity through partial inhibition of the TNF-${\alpha}$ secretion.

• The Journal of the Korea Contents Association
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• v.14 no.6
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• pp.241-246
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• 2014

Acute promyelocytic leukemia (APL) is a cancer of the blood. Although electron beam (EB) irradiation is used with other anti-cancer agents, EB irradiation can be harmful to normal tissues around the cancer. In the present study, we evaluate the differential cytotoxic effect of EB irradiation with other molecules, including TNF-${\alpha}$, on DMSO-treated HL-60 cells and HL-60 cells. HL-60 cells are the human promyleocytic leukemia cell line and are differentiated by DMSO. DMSO-treated HL-60 cells are considered to be normal granulocytic cells. In these results, TNF-${\alpha}$ may be used as the potential agent for the treatment of blood cancer without side effects in low dose of EB irradiation therapy.

• Journal of Physiology & Pathology in Korean Medicine
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• v.30 no.1
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• pp.54-60
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• 2016

Amomum cadamomum Linne (ACL) has long been utilized against the inhibited qi movement related diseases such as dyspepsia, acute gastroenteritis, vomiting and diarrhea in Korean medicine. We speculated that ACL could improve the metabolic disorders such as obesity and type 2 diabetes through removing the phlegm-dampness and promoting the qi movement or stagnation. This study was designed to investigate effects and molecular mechanisms of ACL extract on the improvement of adipocyte dysfunction induced by TNF-α in 3T3-L1 adipocytes. Potential roles of ACL extract in the lipogenesis, inhibition of inflammatory cytokines and insulin resistance, were investigated in this study. Also, we examined the adipose genes and signaling molecules related to insulin resistance and glucose uptake to elucidate its mechanism. Our data demonstrated that TNF-α significantly incresed the release of lipid droplets and the production of MCP-1 and IL-6 from adipocytes. In gene expression, TNF-α reduced the expression of aP2, PPARγ, C/EBPα, GLUT4, and IRS-1 related to lipogenesis and insulin sesitivity, while TNF-α increased the expression of MCP-1 related to inflammation. In addition, TNF-α down-regulated the PPARγ and IRS-1 protein and up-regulated the IRS-1 Ser307 phosphorylation. These alterations induced by TNF-α were prevented by the treatment of ACL extract. Thus, our results indicate that ACL extract can be used to prevent from the TNF-α-induced adipocyte dysfunction through insulin and PPARγ pathways.