• YAKHAK HOEJI
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• v.42 no.6
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• pp.558-566
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• 1998

Two isoforms of cyclooxygenase (COX) have been identified - COX-1, which is constitlitively expressed in most tissues, and the inducible form, COX-2, of which expression is induced by inflammatory signals and mitogens. It has been considered that the beneficial effects of NSAIDs are due to the inhibition of COX-2 activity and the side effects are from the inhibition of COX-1 activity. Therefore, it is essential to develop selective COX-2 inhibitor for developing new GI-tolerable NSAIDS. To discover new leads for developing selective COX-2 inhibitors, three-hundred extracts of natural products were primarily screened with the system of prostaglandin accumulation in LPS-stimulated mouse peritoneal macrophages. To identify whether these inhibitory activities of crude extracts on the accumulation of Prostaglandins were derived from direct action against COX-2, the effects of selected extracts on exogenous arachidonic acid-derived production of prostaglandins by LPS-stimulated macrophages were determined. Among them, 5 methanol extracts of natural products, such as Zingiberis Rhizoma, Alpinae Officinarum Rhizoma, Caryophilli Flos, Scutellariae Radix, Dalbergia ordorifera. inhibited more than 70% of the prostaglandin production in LPS-stimulated mouse peritoneal macrophages at a con-centration of 1${\mu}$g/ml.

• Herbal Formula Science
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• v.19 no.1
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• pp.207-217
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• 2011

Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.342-348
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• 2000

In this experiment, we show the effects of herbal plant extracts on the production of nitric oxide (NO) and TNF-$\alpha$. The extracts of Angelica gigas, Astragalus membranaceus, Acanthopanax sessiliflorus and Houttuynia cordata had no effect on NO synthesis by itself in mouse macrophage cell line (RAW264.7). However, the stimulation with these extracts in the presence of murine interferon-${\gamma}$(mIFN-${\gamma}$) resulted in increased NO synthesis. When these extracts were used in combination with mIFN-${\gamma}$, there were a marked cooperative induction of NO and TNF-$\alpha$ synthesis in a dose-dependent manner. The same results were obtained in the mouse peritoneal macrophages used. The optimal concentration of these extracts on NO synthesis was shown at 100$\mu\textrm{g}$/mL with 100U/mL of mIFN-${\gamma}$. NO synthesis was inhibited by NG-monomethyl-L-arginine. When cell lines were treated with extracts, the expression of inducible NO synthetase (iNOS) was markedly increased in RT-PCR analysis. In addition, synergy between mIFN-${\gamma}$ and extracts was dependent on extracts-induced tumor necrosis factor-$\alpha$(TNF-$\alpha$). These results suggest that water extracts of herbal plants can induce iNOS, NO and TNF-$\alpha$ synthesis of mouse macrophage cell line (RAW264.7) and peritoneal macrophages in combination with mIFN-${\gamma}$.

• Journal of Life Science
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• v.27 no.8
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• pp.873-878
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• 2017

We previously reported that NAD(P)H:quinone oxidoreductase 1 (NQO1)-knockout (KO) mice exhibited spontaneous inflammation in the gut. We also found that NQO1-KO mice showed highly increased inflammatory responses compared with NQO1-WT control mice when subjected to DSS-induced experimental colitis. In a Clostridium difficile toxin-induced mouse enteritis model, NQO1-KO mice were also sensitive compared with NQO1-WT mice. Moreover, numerous studies have shown that NQO1 is functionally associated with immune regulation. Here, we assessed whether NQO1 defects can alter macrophage activation. We found that peritoneal macrophages isolated from NQO1-KO mice produced more IL-6 and $TNF-{\alpha}$ than those isolated from NQO1-WT mice. Moreover, the dicumarol-induced inhibition of NQO1 significantly increased IL-6 and $TNF-{\alpha}$ production in peritoneal macrophages isolated from NQO1-WT mice, as well as in the cultured mouse macrophage cell line, RAW264.7. These results indicate that NQO1 may negatively regulate the activation of macrophages. Knockout or chemical inhibition of NQO1 markedly reduced the expression of $I{\kappa}B$ (inhibitor of $NF{\kappa}B$) in both mouse peritoneal macrophages and RAW264.7 cells. Finally, RAW264.7 cells treated with dicumarol exhibited morphological changes reflecting macrophage activation. Our results suggest that NQO1 may suppress the $NF{\kappa}B$ pathways in macrophages, thereby suppressing the activation of these cells. Thus, immunosuppressive activity may be among the many possible functions of NQO1.

• The Journal of the Korean Society for Microbiology
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• v.7 no.1
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• pp.29-41
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• 1972

To grow Myocbacterium leprae in cultured mouse peritoneal macrophages, studies were made with trypsin-purified Myco. laprae on 1) the dynamics of infection of mouse peritonal macrophages in vivo with Myco. leprae by intraperitoneal inoculation, 2) growth experiment of Myco. leprae in cultured mouse peritoneal macrophages by in vivo infection and in vitro cultivation and 3) the observation of pathological changes in spleens of mice induced by intraperitoneal inoculation of Myco. leprae. Results are summarized as follows; 1. Continuing and significant decreases were observed in the numbers of both acid-fast bacilli in cultured macrophage and of macrophages harboring.acid-fast bacilli by the length of inter vats between the time of intraperitoneal inoculation of Myco. leprae and the time of initiation of macrophage culture. 2. No evidence of multiplication of Myco. leprae in the peritoneal macrophages in vivo was found up to 5 months after intraperitoneal inoculation. 3. With cultures of macrophages made 24 hours and 1 week after intraperitoneal inoculation of Myco. leprae and maintained in vitro up to 2 to 3 months, microscopic examination of the stained preparations of cultured macrophages indicated that an apparent increase in the number of acid-fast bacilli in the macrophages did occur. 4. Quantitative experiment with in vivo infected-in vitro cultured macrophages revealed certain features of increase in the number of total acid-fast bacilli in the cultured macrophages 7 and 9 weeks after initiation of the cultures. 5. Pathological changes in the spleens mice inoculated with Myco. leprae were of mainly degenerative nature in the red pulp. No multiplication of Myco. leprae was observed in the spleens of mice up to 5 months after intraperitoneal inoculation.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.4
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• pp.325-331
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• 2000

Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ plays important roles in inflammatory responses. Some of tetrahydroisoquinoline (THI) compounds exhibited to inhibit iNOS expression in animal studies and RAW 264.7 cells, but the action of THI on inflammatory reaction was not fully investigated. In the present study, we examined a limited series of THIs (higenamine, YS-51 and THI-52) on the $TNF-{\alpha}$ mRNA expression in mouse peritoneal macrophages by Northern analysis. When thioglycollate-stimulated peritoneal macrophages were incubated with LPS (100 ng/ml), expression of $TNF-{\alpha}$ mRNA was evident and reached its maximum at 2.5 h, which was reduced concentration-dependently by treatment with THIs. When the $TNF-{\alpha}$ activity of macrophage-conditioned media was measured using a TNF-sensitive L929 fibroblast cell line, CCL 1, all THIs increased the cell viability in a concentration dependent manner. The concentrations of THIs used are not cytotoxic by itself when analysed by MTT. Furthermore, nitrite/nitrate level was significantly reduced by the presence of THIs in cells treated with $LPS+interferon-{\gamma}\;(IFN-{\gamma}).$ It is concluded, thus, that these results strongly indicated that THIs can suppress the $TNF-{\alpha}$ expression and reduce NO, which may be useful for the inflammatory disorders.

• 대한한약학회:학술대회논문집
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• 2007.11a
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• pp.176-176
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• 2007

• Journal of Haehwa Medicine
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• v.9 no.2
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• pp.315-324
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• 2001

To investigate the capacity of anti-inflammatory cytokines and biological response modifiers (BRMs) to induce IL-$1{\beta}$, IL-6, TNF-${\alpha}$ gene overexpression from mouse macrophages, we isolated the resident peritoneal macrophages from BALB/c mouse (8 week old) and incubated for 6 h with lipopolysaccaride (LPS) and Moschus moschiferus (MOMS) extracts. Analysis of inflammatory cytokines gene expression was carried out by RT-PCR amplification. Amplified PCR products were electrophoresed on 1.2% agarose gel, and the analysis (Ht) was used to 1D-density program. 1. LPS and MOMS extract treatments resulted in a significant decrease in IL-$1{\beta}$, IL-6, TNF-${\alpha}$ mRNA expression level compared with the LPS treatment. 2. Among four sample of MOMS, Inhibitory effects of MOMS-A and MOMS-D for inflammatory cytokines gene expression were to be fine compared with the MOMS-Band MOMS-C. According to the above data, Because the anti- tumoral and anti-inflammatory response activities of macrophage are known to be dependent on the production of inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) by macrophages, we suggest that evaluations of BRM for the reduction of inflammatory cytokines production by macrophages are important for clinical application.

• Journal of Nutrition and Health
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• v.37 no.1
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• pp.23-30
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• 2004

Ginger (Zingiber officinale Roscoe) has been used as a raw material in many traditional preparations since the ancient time. As a component of traditional health products, Ginger is known to be effective as appetite enhancer, anticold and anti-inflammation. This study was performed to investigate the immunomodulative effects of Ginger in mouse, using in vitro and ex vivo experiments. In vitro experiment, the mice splenocytes proliferation and three kinds of cytokines (IL-1 $\beta$, IL-6, and TNF-$\alpha$) prodution by peritoneal macrophages cultured with ethanol and water extracts of Ginger were used to indicate the immunomodulative effect. In order to elucidate the immunomodulative effects of Ginger ex vivo, water extract of Ginger was orally administrated into mice, and isolated splencytes and macrophages were used as experimental model. Ex vivo experiment, six to seven week old mice were fed ad libitum on a chow diet, and water extract of finger was orally administrated every other day for four weeks at two different concentractions (50 and 500 mg/kg B.W./day). In vitro study, the splenocytes proliferation was increased when water extract was supplemented in the range of 50-500 $\mu$l/ml concentration. In case of cytokines production, IL-1 $\beta$, IL-6 and TNF-$\alpha$ released by activated peritoneal macrophages were augmented by the supplementation of water extract of the Ginger. Ex vivo experiment, the highest proliferation of splenocytes and production of cytokines by activated peritoneal macrophages were seen in the mice orally administrated at the concentration of 500 mg/kg B.W./day. In conclusion, this study suggests that Ginger extracts may enhance the immune function by regulating the splenocytes proliferation and enhancing the cytokine prodution capacity by activated macrophages in mice.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.813-818
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• 2002

Expression of monokine induced by IFN-$\gamma$(Mig) mRNA is well-known to strictly depend on Interferon-$\gamma$(IFN-$\gamma$). Lipopolysaccharide (LPS) alone Is weakly effective on Mig mRNA expression in mouse Peritoneal macrophages. This study was undertaken to investigate the synergistic effect of LPS and IFN-$\beta$ on chemokine Mig gene expression in mouse peritoneal macrophages. Although IFN-$\beta$ alone was minimally effective, LPS plus IFN-$\beta$ synergized to produce a high level of Mig mRNh. The synergistic effect of LPS and IFN-$\beta$ (LPS/IFN-$\beta$) on Mig mRNA expression was strain-specific. The most effective synergistic effect of LPS/IFN-$\beta$ on the mRNh expression was found in simultaneous stimulation of LPS/IFN-$\beta$. This synergy was modulated at the level of the gene transcription and was not dependent on a new protein synthesis. Synergistic effect of LPS/IFN-$\beta$ also required the activation of $NF-_KB$. Accordingly, these data suggest that LPS/IFN-$\beta$ synergizes the expression of Mig mRNA through a process that depends on a pretranscriptional level and/or coincident Mig mRNA transcription.