• The Journal of Korean Medicine
• /
• v.30 no.6
• /
• pp.69-79
• /
• 2009

Objective: In this study, the immunomodulatory activity of a mixture of wild Panax ginseng and red-mold rice extracts (MPR) on RAW 264.7 macrophage cells in the presence and absence of methotrexate (MTX), an anti-cancer drug, was investigated. Methods and Results: In the cell viability, MPR showed a significant cell proliferation and inhibited cell regression by red-mold rice (RMR) alone or MTX alone. MPR induced moderate increase in nitric oxide (NO) production. NO production and inducible nitric oxide synthase (iNOS) mRNA expression by LPS decreased after MPR treatment. In addition, MPR slightly induced COX-2 mRNA expression, but it did not affect the expression of COX-2 mRNA by LPS treatment. In RT-PCR analyses, MPR induced IL-$1{\alpha}$, IL-$1{\beta}$, IL-6, and TNF-$\alpha$ mRNA expression, but had no effect on IL-10 and TGF-$\beta$, regardless of MTX treatment. Furthermore, MPR did not interfere with the cytotoxicity of MTX against MCF-7 human breast carcinoma cells. Conclusions: MPR is efficacious in protecting against MTX-induced cell regression as a result of macrophage activation, resulting in induction of cytokine expression, implying that MPR could be considered an adjuvant in MTX-chemotherapy.

• Journal of Life Science
• /
• v.21 no.9
• /
• pp.1259-1265
• /
• 2011

The purpose of this study was to determine whether a single bout of aerobic exercise affects the expression level of toll-like receptor4 (TLR4), IL-6, TNF-${\alpha}$, and suppressor of cytokine signaling-3 (SOCS-3) expression in rat hindlimb muscles depending on fiber types. To accomplish this, thirteen 7-wk Balb/c male mice were randomly assigned to an experimental group or a control group. The exercise protocol consisted of a single bout of treadmill exercise (inclination $10^{\circ}$, speed 17 cm/sec 10 min, 33 cm/sec 10 min, 50 cm/sec) and the animals were killed 24 hr after the exhaustion protocol. The level of TLR4, IL-6, TNF-${\alpha}$, and SOCS-3 mRNA expression was measured by quantitative real-time PCR in soleus and plantaris muscles. A single bout of aerobic treadmill exercise increased TLR4 mRNA expression in the soleus muscle (p<0.05), whereas plantaris TLR4 mRNA expression did not change. Additionally, acute exercise led to a significant increase in IL-6, TNF-${\alpha}$, and SOCS-33 mRNA in the soleus muscle, while transcripts of these genes were not affected by exercise in the plantaris muscle. In conclusion, expression level of several immune-related genes such as TLR4, cytokines, and SOCS-3 is regulated by acute exercise in a fiber type specific manner.

• Journal of Pharmacopuncture
• /
• v.7 no.2
• /
• pp.43-56
• /
• 2004

Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu($BL_{21}$) and Chung- wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant differences. 2. For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3. For survival time, all of experiment groups showed 11.1% increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5. For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6. For expression of cytokine mRNA using RT-PCR, significant increase of IL.-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all off experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effccts in anti-cancer and immune improvement if increasing concentration.

• Korean Journal of Medicinal Crop Science
• /
• v.13 no.5
• /
• pp.213-218
• /
• 2005

Phellinus linteus (PL), one of the immune-regulatory substances, is recognized to play the role in the metabolic process on inflammation and immunity. It has been traditionally used in the oriental medicine to treat inflammatory related disease. The purpose of this study was to evaluate the effects of water extracts of PL on the mesenteric lymph node lymphocytes immune function in the ICR male mice. Control mice received vehicle only. The PL treated mice were administered the respective extract by oral gavages for 4 weeks. IgE concentrations in serum and MLN lymphocytes were significantly lower in PL treated mice than in control mice. PL increased the proportion of $CD4^+\;and\;CD8^+$ T cells in MLN lymphocytes. PL significantly decreased Th2 cytokine concentrations and mRNA expression levels in cytokine secretions. Therefore, water extracts of PL modulate inflammatory parameters through regulation of immunoglobulin production resulting from decreased Th2 cytokine secretion and mRNA expression levels and reduce pro-inflammatory cytokine secretion and mRNA expression in MLN lymphocytes.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.5
• /
• pp.1443-1448
• /
• 2004

The present study was conducted to evaluate the immunomodulatory effects of Dangguijakyak-san(당귀작약산). We investigated the effects of cell proliferation in mouse spleen cell and RAW 264.7 macrophages cells. Dangguijakyak-san enhanced mitogenic activity in the dose-response manner in mouse spleen cells and RAW 264.7 macrophages cells. In nitric oxide (NO) synthesis and iNOS mRNA expression by Dangguijakyak-san, Dangguijakyaksan alone had an effect on NO synthesis and iNOS mRNA expression in RAW 264.7 cells. NO production and iNOS mRNA expression which is excessively induced by LPS decreased after treatment of Dangguijakyak-san. The expressions of cytokine gene by Dangguijakyak-san investigated using reverse transcription polymerase chain reaction (RT-PCR). In RT-PCR, IL-1α, IL-1β and IL-6 mRNA expressions induced in Dangguijakyak-san-treated RAW 264.7 cells. These data indicate that 1) Dangguijakyak-san can modulate various immune response and 2) the immunomodulatory effects of Dangguijakyak-san may be, in part, associated with the regulation of NO synthesis, the expressions of these cytokine as well as the mitogenic effect on spleen cells and macrophages cells.

• Korean Journal of Veterinary Pathology
• /
• v.3 no.1
• /
• pp.15-25
• /
• 1999

To elucidate the mechanism of age-related development in FGS/NgaKIST mice with spontaneous glomerulosclerotic lesion, we examined expression and localization of various cytokine mRNA in the kidney in the progression of diseases. This mouse model is the first to develop spontanously occuring glomerosclerotic lesion in the kidney. In this study, we detected the up-regulation of local cytokine genes such as IL-1$\beta$, IL-2, IL-6, IL-10, TNF-$\alpha$, TGF-$\beta$, and IFN- $\gamma$ in the kidneys. In RT-PCR and Southern blot analysis, we detected gradual expressions of cytokine mRNA of IL-1$\beta$, IL-2, IL-6, IFN- $\gamma$, and TNF $\alpha$ mRNA during the course of disease. Other cytokines including IL -10 and TGF -$\beta$ were found to be appeared the slightly expressed level at 3 to 12 weeks before onset of inflammatory lesion but they are highly expressed at the end-stage of the disease accompaning high proteinurea and wasting. In situ RT-PCR, each cytokine mRNA were specifically localized in a variety of cells including mesangial, endothelial, parietal epithelial, tubular epithelial, arterial muscle cell, and infiltrated inflammatory cells. In addition, TNF - $\alpha$was detected moderately in the visceral and parietal epithelial cell, but weakly in endothelial and mesangial cells, whereas IL-1 $\beta$ and IL -6 were strong in mesangial regions. IL-6 and TNF- $\alpha$ was highly localized in the damaged proximal and collecting tubules. Especially, TGF -$\beta$ mRNA was highly found in mesangial cells within glomerulus and interstitium during the end-stage of this disease.. These results indicate that pro inflammatory cytokines such as IL-1 $\beta$, IL-2, IL-6, and TNF- $\alpha$ were gradually expressed from the early stage of this disease to the end-stage, and that IL-10 and TGF-$\beta$ may be important in the accumulation of extracellular matrix(ECM) within glomerulus and periglomerular fibrosis in the progression of this disease as well as tissue destruction in end-stage of this disease.

• Tuberculosis and Respiratory Diseases
• /
• v.47 no.2
• /
• pp.172-183
• /
• 1999

Background: Nitric oxide is a short-lived effector molecule derived from L-arginine by the nitric oxide synthase(NOS). Nitric oxide plays a role in a number of physiologic and pathophysiologic functions including host defense, edema formation, and regulation of smooth muscle tone. Some kinds of cells including macrophage are known to produce large quantities of nitric oxide in response to inflammatory stimuli such as interleukin-$1\beta$(IL-$1\beta$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), interferon-$\gamma$(IFN-$\gamma$) and lipopolysaccharide(LPS). Reactive oxygen species are also known to be important in the pathogenesis of acute cell and tissue injury such as acute lung injury model Methods: Using the RA W264.7 cells, we have examined the ability of oxidant hydrogen peroxide($H_2O_2$) to stimulate nitric oxide production and inducible NOS mRNA expression. Also, we have examined the effects of NOS inhibitors and antioxidants on $H_2O_2$ induced nitric oxide production. Results: Stimulation of RAW264.7 cells with combinations of 100 ng/ml IL-$1\beta$, 100 ng/ml TNF-$\alpha$, and 100 U/ml IFN-$\gamma$ or 100 U/ml IFN-$\gamma$ and $1{\mu}g/ml$ LPS induced the synthesis of nitric oxide as measured by the oxidation products nitrite($NO_2^-$) and nitrate($NO_3^-$). Addition of $250 {\mu}M-2$ mM $H_2O_2$ to the cytokines significantly augmented the synthesis of $NO_2^-$ and $NO_3^-$(p<0.05). When cells were incubated with increasing concentrations of $H_2O_2$ in the presence of IL-$1\beta$, TNF-$\alpha$ and IFN-$\gamma$ at constant level, the synthesis of $NO_2^-$ and $NO_3^-$ was dose-dependently increased(p<0.05). $N^G$-nitro-L-arginine methyl ester(L-NAME), dose dependently, significantly inhibited the formation of $NO_2^-$ and $NO_3^-$ in cells stimulated with LPS, IFN-$\gamma$ and $H_2O_2$ at constant level(p<0.05). Catalase significantly inhibited the $H_2O_2$-induced augmentation of cytokine-induced $NO_2^-$ and $NO_3^-$ formation(p<0.05). But, boiled catalase did not produce a significant inhibition in comparison with the native enzyme. Another antioxidant 2-mercaptoethanol and orthophenanthroline dose-dependently suppressed $NO_2^-$ and $NO_3^-$ synthesis(p<0.05). Northern blotting demonstrated that H:02 synergistically stimulated the cytokine-induced iNOS mRNA expression in RA W264.7. Conclusion: These results suggest that $H_2O_2$ contributes to inflammatory process by augmenting the iNOS expression and nitric oxide synthesis induced by cytokines.

• Korean Journal of Food Science and Technology
• /
• v.36 no.6
• /
• pp.986-990
• /
• 2004

The immunomodulatory effect of Codonopsis lanceolata based on the production of cytokines and the activation of macrophage was studied. The mRNA expression of nitric oxide synthase (iNOS) was gradually induced after 24 hr treatment of Codonopsis lanceolata, and NO production was a maximum after 24 hr treatment with 1 mg/mL. RAW 264.7 cell on in vitro treatment with Codonopsis lanceolata induced mRNA of cytokines such as interleukin-1(IL-1)${\beta}$, interleukin-6(IL-6), tumor necrosis $factor(TNF)-{\alpha}\;and\;interferon(IFN)-{\gamma}$; $IL-1{\beta}$ and IL-6 mRNA were gradually induced up to 24 hr, $TNF-{\alpha}\;mRNA$ was regularly induced up to 24 hr, and $IFN-{\gamma}\;mRNA$ level was a maximum within 1 hr. These results suggest that Codonopsis lanceolata exerts as an effective immunomodulator and enhances antitumor activity of macrophages.

• The Journal of Korean Medicine
• /
• v.24 no.3
• /
• pp.84-95
• /
• 2003

Objective : This study was carried out to investigate the effects of Kamisopunghwalhyeol-tang (Jiaweishujenghuoxie-tang; Kami-SPHHT) on the immunity responses of the Synoviocytes isolated from the patients on rheumatoid arthritis. Methods : Cells were stimulated by $Interleukin-1{\beta}$ and Tumor Necrosis $Factor-{\alpha}$ in the presence or absence of Kami-SPHHT, and then induced cytokine mRNA levels were determined by RT-PCR and real-time quantitative RT-PCR. Results : Levels of $IL-1{\beta},{\;}IL-6,{\;}TNF-{\alpha}$, COX-2, and NOS II mRNA expressions significantly decreased in Kami-SPHHT treated cells compared to non-treated control cells. Also, DNA-binding activity of $NF-{\kappa}B$ and AP-l decreased in Kami- SPHHT treated hFLSs. Conclusion : These results suggest that Kami-SPHHT may be involved in anti-inflammatory reactions by inducing cytokine gene expression in synoviocytes, and further in vivo examination on its efficacy can provide potential application for the treatment of rheumatoid arthritis.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.26 no.6
• /
• pp.915-921
• /
• 2012

Artemisiae Asiaticae Herba (AAH) has been used to remedy of symptoms such as bleeding, dysmenorrhea, eczema and itchy skin in Oriental Medicine. In this study, we investigated the protective effect of AAH on allergic response. The effect of AAH was analyzed by ELISA and RT-PCR in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase and histamine as markers of degranulation, production of IL-4 and TNF-${\alpha}$, and gene expression of HDC2, cytokines and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit. We found that AAH suppressed ${\beta}$-hexosaminidase and histamine release, the production of IL-4 and TNF-${\alpha}$ in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. AAH also significantly decreased cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12, IL-13, TNF-${\alpha}$, and GM-CSF, and increased cytokine mRNA expressions of IL-10 in RBL-2H3. In addition, AAH suppressed mRNA expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit on cell surface. Our results indicate that AAH protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit.