• Journal of Yeungnam Medical Science
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• v.20 no.2
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• pp.129-141
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• 2003

Background: Leptin is a 16-KDa non-glycosylated peptide hormone synthesized almost exclusively by adipocytes. The well-known function of leptin is regulation of food intake and energy expenditure. Leptin also plays a regulatory role in immune and inflammatory process including cytokine production. The purpose of this study was to investigate the effect of leptin on the expression of several chemokine genes(RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$) in THP-1 cells. Materials and Methods: Total RNA of THP-1 cells were prepared by Trizol method, and then stimulated with the leptin(250 ng/$m{\ell}$) or LPS(100 ng/$m{\ell}$). We examined the expression patterns of various chemokine mRNAs in THP-1 cell lines by RT-PCR and Northern blot. Results: Leptin did not induce the expression of chemokine mRNAs in THP-1 cells. The expression patterns of RANTES, IL-8, MCP-1, IP-10, and Mig mRNAs in THP-1 cells stimulated with leptin and LPS simultaneously was almost same to the patterns of LPS alone-induced chemokine mRNAs. RANTES mRNA expression was independent on the concentrations of leptin. Although leptin did not have strong effect on the expression of RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$ mRNAs in THP-1 cells, leptin could induce the expression of long isoform of leptin receptor(OB-RL) mRNA, and its expression was elevated in simultaneous stimulation of leptin and LPS. Conclusion: These data suggest that leptin is able to induce OB-RL in THP-1 cells, however, leptin has little effect on the expression of pro-inflammatory chemokine genes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.779-784
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• 2005

Mast cells are a potent source of mediators that regulate the inflammatory response in allergic reaction. To evaluate the atopic allergic effect of Asiasari radix(AR), the author investigated a possible effect of AR on mast cell-mediated allergic reaction, cytokines secretion and mRNA expression in vivo and in vitro. In mice orally administered by AR (0.01, 0.1 and 1.0 g/kg) for 1 h, compound 48/80-induced ear swelling was significantly reduced (P < 0.05). AR had inhibitory effects on tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-8, and IL-6 secretion from HMC-1. Significant reduced levels of $TNF-{\alpha}$ mRNA was observed in the human mast cell line (HMC-1) with AR (1.0 mg/ml). In addition, AR had no cytotoxic effect on cell viability. These results suggest that AR contributes to the treatment of atopic allergic reactions, and that its action may be due to inhibition of cytokine secretion and mRNA expression in HMC-1.

• The Journal of Korean Medicine
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• v.27 no.4
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• pp.1-11
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• 2006

Objectives : We investigated the effect of Dohongsamul-tang (DHSMT) on the production of various cytokines in lipopolysaccaride (LPS) stimulated peripheral mononuclear cells (PBMCs) from CI patients. Methods: Cell viability was determined using MTT assay. ELISA was carried out for investigating $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, IL-8, IL-4, and $TGF-{\beta}$ 1 Results : The amount of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, IL-6, IL-8, IL-4, and transforming growth factor(TGF)-${\beta}$ 1 in PBMC culture supernatant significantly increased in the LPS treated cells compared to unstimulated cells. We show that DHSMT inhibited the production of TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 induced by LPS in a dose dependent manner. The maximal inhibition rate of the TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 production by pretreatment of DHSMT (1.0mg/ml) was 38.52 ${\pm}$ 2.5% (P < 0.01), 44.02 ${\pm}$ 3.5% (P < 0.05), 45.32 ${\pm}$ 2.3% (P < 0.01), 42.30 ${\pm}$ 3.1% (P < 0.05), and 49.70　${\pm}$　3.1%(P ＆lt; 0.05), respectively. On the other hand, DMSMT significantly increased the LPS-induced TGF-${\beta}$ 1 production(P<0.05). Conclusions : Taken together. these results suggest that DHSMT might have regulatory effects on cytokine production, which might explain its beneficial effect in the treatment of CI.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.902-908
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• 2006

Tetrandra is the root of Stephania tetrandra 5. Moore (family Menispermaceae), or of Aristolochia frangchi Wu (family Aristolochiaceae). It is a Differ-flavored and cold-property herb acting on the urinary bladder, kidney and spleen meridiands. Known biological effects of this herb are expelling wind to relieve pain and inducing diuresis to alleviate edema. This herb also has anti-inflammatory and anti-hypersensitivity actions. Recent studies have shown that Stephanniae Tetrandrae Radix has antimicrobial effects, namely, a protective effect on acute renal failure induce by gentamicin sulfate and a suppressive effect against clostridium perfringes. However, there is a lack of studies concerning the immunological activities of this herb. The present study was conducted to evaluate the immunological activities of Stephanniae Tetrandrae Radix on the regulatory mechanisms of cytokines and nitric oxide (NO) in Raw 264.7 cells. Cell viability was measured by MTT assay after the treatment of Stephanniae Tetrandrae Radix extract (STRE) and NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidences that STRE inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$ and interleukin-6 (IL-6) in Raw 264.7 cells activated with lipopolysaccharide (LPS). These findings showed that STRE could produce some anti-inflammatory effects which might play a role in adjunctive therapy in Gram-negative bacterial infections.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.11-18
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• 2008

SunBangHwalMyungEum (SBH) has the effects of subduing swelling, resolving masses and alleviating pain in traditional oriental medicine. Recent studies showed that SunBangHwalMyungEum produced anti-cancer, anti-metastasis and immuno-modulatory effects. However there is lack of studies regarding the effects of SBH on the immunological activities. The present study was conducted to evaluate the effect of SBH on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Methods : After the treatment of SBH, 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. Results : Results provided evidence that SBH inhibited the production of NO, iNOS, $interleukin-1{\beta}$ ($(IL-1{\beta})$), IL-6, and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with lipopolysaccharide. Conclusions : These findings suggest that SBH can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• IMMUNE NETWORK
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• v.23 no.6
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• pp.48.1-48.21
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• 2023

Mesenchymal stromal/stem cells (MSCs) possess immunoregulatory properties and their regulatory functions represent a potential therapy for acute lung injury (ALI). However, uncertainties remain with respect to defining MSCs-derived immunomodulatory pathways. Therefore, this study aimed to investigate the mechanism underlying the enhanced effect of human recombinant bone morphogenic protein-2 (rhBMP-2) primed ES-MSCs (MSC^BMP2) in promoting Tregs in ALI mice. MSC were preconditioned with 100 ng/ml rhBMP-2 for 24 h, and then administrated to mice by intravenous injection after intratracheal injection of 1 mg/kg LPS. Treating MSCs with rhBMP-2 significantly increased cellular proliferation and migration, and cytokines array reveled that cytokines release by MSC^BMP2 were associated with migration and growth. MSC^BMP2 ameliorated LPS induced lung injury and reduced myeloperoxidase activity and permeability in mice exposed to LPS. Levels of inducible nitric oxide synthase were decreased while levels of total glutathione and superoxide dismutase activity were further increased via inhibition of phosphorylated STAT1 in ALI mice treated with MSC^BMP2. MSC^BMP2 treatment increased the protein level of IDO1, indicating an increase in Treg cells, and Foxp3⁺CD25⁺ Treg of CD4⁺ cells were further increased in ALI mice treated with MSC^BMP2. In co-culture assays with MSCs and RAW264.7 cells, the protein level of IDO1 was further induced in MSC^BMP2. Additionally, cytokine release of IL-10 was enhanced while both IL-6 and TNF-α were further inhibited. In conclusion, these findings suggest that MSC^BMP2 has therapeutic potential to reduce massive inflammation of respiratory diseases by promoting Treg cells.

• Journal of Life Science
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• v.23 no.1
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• pp.38-43
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• 2013

The objective of this study was to evaluate the effect of the synthetic CopA3 peptide of Copris tripartitus on skin inflammation. Regulatory mechanisms of cytokines and nitric oxide (NO) are involved in the immunological activity of RAW 264.7 cells. Tested cells were treated with different concentrations of CopA3 and further cultured for an appropriate time after lipopolyssacharide (LPS) addition. During the entire experimental period, 5, 25, 50, and 100 ${\mu}g/ml$ of CopA3 had no cytotoxicity. At these concentrations, CopA3 inhibited tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). CopA3 also inhibited the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). CopA3 inhibited the activity of iNOS and COX-2 by 41% and 59%, respectively, at 100 ${\mu}g/ml$. In addition, CopA3 reduced the release of inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These results suggest that CopA3 may have significant effects on inflammatory factors and that it may be a potential anti-inflammatory therapeutic agent.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.117-125
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• 2007

Objectives : Berberine, a main alkaloid component of Coptidis rhizoma, has an antimicrobial and anti-tumor activities and antiinflammatory effect. In the present study, we investigated effect of berberine on the production of inflammatory mediators such as nitric oxide(NO), prostaglandin E2(PGE2), TNF-${\alpha}$ and IL-1${\beta}$ in LPS-stimulated BV2 microglial cells, Methods : BV2 cells were pre-treated with berberine and then stimulated with LPS. The cytotoxicity of berberine was determined by MTT assay. The NO production was measured by Griess assay. The mRNA expression and protein levels of inducible nirtic oxide synthase(iNOS) were determined by RT-PCR and Western blot. The production of PGE2 and cytokines was measured by ELISA. Results : Berberine inhibited the production of NO, PGE2 and pro inflammatory cytokines, TNF-${\alpha}$ and IL-1${\beta}$ in a dose dependent manner in LPS-stimulated BV2 cells. In addition, berebrine greatly suppressed the mRNA expression and protein levels of iNOS and inflammatory cytokines induced by LPS stimulation. These results indicate that the post-transcriptional regulatory mechanism of iNOS and/or inflammatory cytokine gene expression by berberine is involved in its anti-inflammatory effects, respectively. Conclusion : The present study suggests that berberine can be useful as a potential anti-inflammatory agent for treatment of various neurodegenerative diseases such as Alsheimer's disease, Parkinson's disease and stroke.

• IMMUNE NETWORK
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• v.15 no.3
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• pp.150-160
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• 2015

We previously reported that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) accelerates hematopoiesis and has an improving effect on animal disease models such as sepsis and asthma. The effects of PLAG supplementation on immune modulation were assessed in healthy men and women. The objective was to evaluate the effects of PLAG supplementation on immune regulatory functions such as activities of immune cells and cytokine production. A randomized double blind placebo-controlled trial was conducted. Seventy-five participants were assigned to one of two groups; all participants had an appropriate number of white blood cells on the testing day. The PLAG group (n=27) received oral PLAG supplements and the control group (n=22) received oral soybean oil supplements. IL-4 and IL-6 production by peripheral blood mononuclear cells (PBMC) were lower (p<0.001 and p<0.001, respectively) with PLAG than with soybean oil. However, the production of IL-2 and IFN-$\gamma$ by PBMC was unaltered with PLAG supplementation. The B cell proliferation decreased significantly in the PLAG group compared to the soybean oil control (p<0.05). The intake of PLAG in healthy adults for 4 weeks was deemed safe. These data suggest that PLAG has an immunomodulatory function that inhibits the excessive immune activity of immunological disorders such as atopic and autoimmune diseases. PLAG could improve the condition of these diseases safely as a health food supplement.

• Animal cells and systems
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• v.10 no.1
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• pp.15-20
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• 2006

Allergic inflammation is thought to be a Th2 cell-dominant immune response during which tissue-resident fibroblasts produce chemokines which contribute to the recruitment of migratory leukocytes to sites of tissue injury. Thymus and activation-regulated chemokine (TARC; CCL17) is a potent member of the CC chemokine family and a selective chemoattractant for Th2 cells. In order to study the regulatory profiles of TARC production by $TNF-{\alpha}$, $IFN-{\gamma}$, and Il-4 in human normal skin fibroblast, CCD-986sk cell line was used. The expression of TARC protein was measured using ELISA, and mRNA level was detected by RT-PCR. The combination of $TNF-{\alpha}$ and IL-4 induced a time-and dose-dependent synergistic increase in the expression of TARC at both protein and mRNA levels in the cultured human skin fibroblasts. Exposure of the cells to single cytokine had no effect on TARC expression. The high concentration (100 ng/ml) and long incubation time (72 h) of $IFN-{\gamma}$ further enhanced the TARC production induced by $TNF-{\alpha}$/lL-4 in the skin fibroblast. This synergistic effect of Th1 and Th2 type cytokines on TARC production by skin fibroblasts may contribute to the inflammatory cell infiltration and tissue damage with allergic inflammation.