• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.254-259
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• 2004

Jeongshin-tang (JST) is a Korean herbal prescription, which has been successfully applied for the various neuronal diseases. However, it's effect remains unknown in experimental models. To investigate the biological effect of JST in Alzheimer's disease (AD) in vitro model, we analized the production of interleukin (IL)-6 and IL-8, and expression of cyclooxygenase (COX)-2 in IL-1β plus β-amyloid [25-35] fragment (A)-stimulated human astrocytoma cell line U373MG. JST alone had no effect on the cell viability. The production of IL-6 and IL-8 was significantly inhibited by pretreatment with JST (1mg/㎖) on IL-1β plus A-stimulated U373MG cells. Maximal inhibition rate of IL-6 and IL-8 production by JST was about 41.22% (P<0.01) and 34.45% (P<0.05), respectively. The expression level of COX-2 protein was up-regulated by IL-1β plus A but the increased level of COX-2 was inhibited by pretreatment with JST (1 mg/㎖). These data indicate that JST has a regulatory effect on cytokine production and COX-2 expression, which might explain it's beneficial effect in the treatment of AD.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.1044-1050
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• 2006

The Korean genuine medicine, 'Gigukjihwangtanggami (GJT)' has long been used clinically for hypertension and various cerebrovascular diseases. However, experimental study has been carried out very little. Recently cytokines involved in the regulation of inflammatory reactions and immune responses may play an important role in the pathogenesis of cerebral infarction (CI). The aim of this study is to investigate the effect of GJT on the production of pro-inflammatory cytokines and anti-inflammatory cytokines in peripheral blood mononuclear cells (PBMCS) stimulated with lipopolysaccaride (LPS) from Cl patients. The amount of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha}),\;interleukin-1{\beta}\;(IL-1{\beta})$, IL-6, IL-8, IL-4 and IL-10 in PBMC culture supernatant was significantly increased in the LPS treated cells compared to unstimulated cells. GJT inhibited the production of $TNF-{\alpha},\;IL-1{\beta}$, IL-6 and IL-8 induced by LPS. The maximal inhibition rate of the $TNF-{\alpha},\;IL-1{\beta}$, IL-6 and IL-8 production by pretreatment of GJT (1 mg/ml) was $57.32{\pm}2.5%$ (p.0.05), $42.02{\pm}3.5%$ (p.0.05), $40.02{\pm}2.3%$ (p.0.05) and $48.02{\pm}3.1$, (p.0.05), respectively. In the Other hand, GJT increased the production of IL-4 and IL-10. The maximal increase rate of the IL-4 and IL-10 production by pretreatment of GJT (1 mg/ml) was $42.4{\pm}3.3%$ (p.0.05) End $56.4{\pm}2.9%$ (p.0.05), respectively. Taken together, these results indicate that GJT Ray have regulatory effects on the cytokine production and suggest that GJT might use clinically for the treatment of CI.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.721-727
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• 2007

Scorpion (SCP) has been clinically used for the treatment of endogenous wind to relieve convulsion, clearing away toxins, resolving hard masses and removing obstruction in the collaterals to relieve pain. Recent studies showed that scorpion toxins that affect the activating mechanism of sodium channels and indian black scorpion venom induced anti-proliferative and apoptogenic activity against human leukemic cell lines U937 and K562. There is lack of studies regarding the effects of SCP on the immunological activities. The present study was conducted to evaluate the effect of SCP on the regulatory effects of cytokines and nitric oxide (NO) for the immunological activities in Raw 264.7 cells. After the treatment of SCP MeOH extract dissolved in media for 1 h prior to the addition of lipopolysaccharide (LPS: 1 ${\mu}$g/ml), cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. As results, SCP inhibited the production of nitrite and nitrate (0.3 and 1.0 mg/ml), iNOS and p-$I_KB_{\alpha}$ protein, tumor necrosis factor-${\alpha}$ (0.3 and 1.0 mg/ml), interleukin-1${\beta}$ (0.3 and 1.0 mg/ml) and interleukin-6 (1.0mg/ml) in Raw 264.7 cells activated with LPS. These findings suggest that SCP can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• The Journal of Pediatrics of Korean Medicine
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• v.18 no.2
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• pp.61-76
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• 2004

Objective : To demonstrate of regulatory effect of atopic allergic regulation by Chungsangbangpoong-Tang(CBT), This experiment was studied. Methods : The author investigated a possible effect of CBT on cytokines production using human T cell line (MOLT-4) or human mast cell line (HMC-1). In addition, the author investigated whether CBT has inhitory effects on compound 48/80- induced histamine release from rat peritoneal mast cells (RPMC) and compound 48/80-induced ear swelling in ICR mice. Results : CBT (0.01 mg/ml)-containing medium in stimulated culture supernatants significantly increased IL-2 secretion compared with untreated MOLT-4, whereas CBT (0.01-1.0 mg/ml)-containing medium in stimulated culture supernatants significantly decreased IL-4 secretion compared with untreated MOLT-4. Significant reduced levels of IL -6 and $TNF-{\alpha}$ were observed in the HMC-l with CBT (P<0.05). CBT did not inhibit the histamine release from the RPMC but inhibit ear swelling response. Conclusion : These results suggest that CBT contributes to the treatment of atopic allergic reactions, such as atopic dermatitis and that its action may be due to regulation of cytokine production.

• BMB Reports
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• v.48 no.3
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• pp.172-177
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• 2015

Upregulation of pro-inflammatory mediators contributes to ${\beta}$-cell destruction and enhanced infiltration of immune cells into pancreatic islets during development of type 1 diabetes mellitus. In this study, we examined the regulatory effects and the mechanisms of action of celastrol against cytotoxicity and pro-inflammatory immune responses in the RINm5F rat pancreatic ${\beta}$-cell line stimulated with a combination of interleukin-1 beta, tumor necrosis factor-alpha, and interferon-${\gamma}$. Celastrol significantly restored cytokine-induced cell death and significantly inhibited cytokine-induced nitric oxide production. In addition, the protective effect of celastrol was correlated with a reduction in pro-inflammatory mediators, such as inducible nitric oxide synthase, cyclooxygenase-2, and CC chemokine ligand 2. Furthermore, celastrol significantly suppressed cytokine-induced signaling cascades leading to nuclear factor kappa B (NF-${\kappa}B$) activation, including $I{\kappa}B$-kinase (IKK) activation, $I{\kappa}B$ degradation, p65 phosphorylation, and p65 DNA binding activity. These results suggest that celastrol may exert its cytoprotective activity by suppressing cytokine-induced expression of pro-inflammatory mediators by inhibiting activation of NF-${\kappa}B$ in RINm5F cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.2
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• pp.491-497
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• 2007

Hyeonggaeyeongyo-tang (HYT; Jingjielianqiao-tang), is known to be effective in lowering wind-heat blended as a pathogen of kidney. HYT has been traditionally used for the treatment of a syndrome in kidney meridian, due to invasion of pathogenic wind and heat. Nowadays, this prescription is used to treat diseases marked by excessive wind and heat in the kidney meridian, such as acute otitis media, empyema, hypertrophic rhinitis, nasal bleeding, nasal obstruction, acne and tonsillitis. The present study was conducted to evaluate the effect of HYT on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in Raw 264.7 cells. After the treatment of HYT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX-2_ and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. The production of No was significantly inhibited by pre-treatment (1h) with HYT(0.1-0.3 mg/ml) on LPS-activated Raw264.7 cells. The expression of iNOS and COX-2 protein were up-regulated by LPS, but the increased levels of iNOS and COX-2 were inhibited by pre-treatment of HYT (0.3-1.0 mg/ml), respectively. And the level of interleukin-6 (IL-6), cytokine released from macrophage, was reduced by HYT pre-treatment (0.3-1.0 mg/ml). Thus, the present data suggest that HYT may play an important role in adjunctive therapy in Gram-negative bacterial infections.

• Journal of fish pathology
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• v.34 no.1
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• pp.23-29
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• 2021

Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 of zebrafish (Danio rerio) was negative regulation of the interferonΦ1 and 3 response in vitro. In this study, we analyze the induction of in vivo immune response activation from the IRF10 gene of zebrafish and the protective effect against VHSV. As the results, the group inoculated with IRF10 expression vectors, there was no expression of IFNΦ1, suggestion that IRF10 may function as a negative regulator of IRF3, which binds to the IFNΦ1 promoter. And other types of interferon genes (IFNΦ2-4) are thought to have been activated, inducing to the expression of pro-inflammatory cytokine and Mx genes. As the results of challenge test performed at 14 days after inoculation of the expression vectors, the maximum survival rate [50% (1㎍ DNA) and 42.5% (10㎍ DNA)] for IRF10 group were recorded. Meanwhile, the survival rates of pcDNA3.1 and PBS as the control groups were 10% and 15%, respectively. This study suggests that the possibility that activation of IRF10 molecule could be exploited as a VHS control method.

• Advances in Traditional Medicine
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• v.7 no.3
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• pp.321-329
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• 2007

The present study was conducted to evaluate the effect of Yongdamsagantang (YST) on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in RAW 264.7 cells. After the treatment of YST water extract, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) and phospholylation of inhibitor of nuclear factor kappa B alpha ($p-I{\kappa}B{\alpha}$) were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidences that YST inhibited the production of NO. iNOS, and interleukin-6, and the activation of $p-I{\kappa}B{\alpha}$ in RAW 264.7 cells activated with lipopolysaccharide. These findings showed that YST could have some anti-inflammatory effects which might play a role in therapy in Gram-negative bacterial infections.

• The Journal of Internal Korean Medicine
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• v.31 no.2
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• pp.189-200
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• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.459-465
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• 2011

The nuclear factor of activated T cells (NFAT) protein induces transcriptions of cytokine genes including IL-2 for T-cell activation. Normally activation of NFAT is important to induce immune responses but excessive NFAT activation provokes immunopathological reactions such as autoimmunity, transplant rejection, and inflammation. Thus, for the treatment of autoimmune diseases drugs repressing the activation of NFAT have been searched. In this study, immnunosuppressive effects of Rosa chinensis Jacq. extracts identified as a potent NFAT inhibitor from a natural product library were examined. NFAT reporter assay, MTS assay, real time PCR, IL-2 ELISA, MLR, and FACS (Fluorescent Activated Cell Sorting) were used to measure inhibitory immunocyte activities of Rosa chinensis Jacq. The variety of natural products have been screened and some were found to show inhibitory activities against the NFAT transcription factor. Among them, extract of Rosa chinensis Jacq. showed an strong inhibitory effect on the activation of NFAT without affecting cell viability. Levels of IL-2 transcripts as well as IL-2 protein were decreased with treatment of Rosa chinensis Jacq. extract. In addition, immunosuppressive activity of Rosa chinensis Jacq. extract was exhibited in the mixed leukocytes reaction. The increasement of CD4+CD25+ (Treg) immunocyte was also detected in the analysis using FACS after applying Rosa chinensis Jacq. extract. Immunosuppressive effects of the Rosa chinensis Jacq. extracts were clearly demonstrated in the present study. In addition, Rosa chinensis Jacq. extract also positively affected regulatory T cell induction. Further investigations in particular on purification of single substance responsible for the immunosuppressive effects from the extract and analysis on possible actions of the extract in interfering cell signaling and cytokine production will be needed.