• IMMUNE NETWORK
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• v.12 no.5
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• pp.207-212
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• 2012

T cell immunoglobulin mucin domain (TIM)-3 is an immunomodulatory molecule and upregulated in T cells by several cytokines. TIM-3 also influences mast cell function but its transcriptional regulation in mast cells has not been clarified. Therefore, we examined the transcript level and the promoter activity of TIM-3 in mast cells. The TIM-3 transcript level was assessed by real-time RT-PCR and promoter activity by luciferase reporter assay. TIM-3 mRNA levels were increased in HMC-1, a human mast cell line by TGF-${\beta}1$ stimulation but not by stimulation with interferon (IFN)-${\alpha}$, IFN-${\lambda}$, TNF-${\alpha}$, or IL-10. TIM-3 promoter -349~+144 bp region relative to the transcription start site was crucial for the basal and TGF-${\beta}1$-induced TIM-3 promoter activities in HMC-1 cells. TIM-3 promoter activity was increased by over-expression of Smad2 and Smad4, downstream molecules of TGF-${\beta}1$ signaling. Our results localize TIM-3 promoter activity to the region spanning -349 to ＋144 bp in resting and TGF-${\beta}1$ stimulated mast cells.

• Parasites, Hosts and Diseases
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• v.52 no.4
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• pp.355-365
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• 2014

The enteric protozoan parasite Entamoeba histolytica is the causative agent of human amebiasis. During infection, adherence of E. histolytica through Gal/GalNAc lectin on the surface of the amoeba can induce caspase-3-dependent or -independent host cell death. Phosphorylinositol 3-kinase (PI3K) and protein kinase C (PKC) in E. histolytica play an important function in the adhesion, killing, or phagocytosis of target cells. In this study, we examined the role of amoebic PI3K and PKC in amoeba-induced apoptotic cell death in Jurkat T cells. When Jurkat T cells were incubated with E. histolytica trophozoites, phosphatidylserine (PS) externalization and DNA fragmentation in Jurkat cells were markedly increased compared to those of cells incubated with medium alone. However, when amoebae were pretreated with a PI3K inhibitor, wortmannin before being incubated with E. histolytica, E. histolytica-induced PS externalization and DNA fragmentation in Jurkat cells were significantly reduced compared to results for amoebae pretreated with DMSO. In addition, pretreatment of amoebae with a PKC inhibitor, staurosporine strongly inhibited Jurkat T cell death. However, E. histolytica-induced cleavage of caspase-3, -6, and -7 were not inhibited by pretreatment of amoebae with wortmannin or staurosporin. In addition, we found that amoebic PI3K and PKC have an important role on amoeba adhesion to host compartment. These results suggest that amebic PI3K and PKC activation may play an important role in caspase-independent cell death in Entamoeba-induced apoptosis.

• Journal of Korean Medicine for Obesity Research
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• v.7 no.2
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• pp.39-44
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• 2007

Objectives In recent years, we are concerned about anti-aging and longevity. And I heard that Mori Fructus has been used for promotion of health in local clinics. So I wanted to know immuno-ability of Mori Fructus and carried out this experiment. Methods We cultivated 3T3-L1 Preadipocytes and Protein chip used for $ProteoPlex^{TM}$ 16-Well Murine Cytokine Array Kit. Results We known the immunity of Mori Fructus about 3T3-L1 Preadipocytes and gained the increase of Cytokines(IL-2, IL-4, IL-12p70, GM-CSF, INF-${\gamma}$, TNF-${\alpha}$). Conclusions So I guess that Mori Fructus has effect of immuno-ability, etc.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.292-292
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• 2017

This study was conducted to evaluate the effect of anti-obesity and antioxidant enzyme activities in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The cytotoxicity of different solvent fractions of C. lanceolata on 3T3-L1 preadipocytes were evaluated using the MTT assay, the rate of cell survival progressively decreased in a dose-dependent manner. Butyl alcohol fraction at $200{\mu}g/mL$ exhibited a pronounced cytotoxic effect (75.73%) on 3T3-L1 cell comparable to that of the hexane fraction (79.82%), methylene chloride fraction (84.02%), ethyl acetate fraction (87.62%) and DW fraction (86.30%) at the same concentration. The Oil Red O solution was used to determine whether different solvent fractions of C. lanceolata induce adipocyte differentiation in 3T3-L1 preadipocytes. Confluent 3T3-L1 cells were treated with $50{\mu}g/mL$ concentration of solvent fraction extracts from C. lanceolata. Inhibitory degree of lipid accumulation against solvent fraction extracts showed a significant level compared with the control. Both lipid accumulation and adipocyte differentiation showed relatively high effect on methyl chloride fraction. The root extract of C. lanceolata had the highest SOD enzyme activity of 84.5% in ethyl acetate partition layer and while water partition layer of diploid showed the lowest SOD enzyme activity of 57.9%. The activity of CAT, APX and POD showed a significantly higher activity in ethyl acetate partition layer compared with the other fraction. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on anti-obesity and antioxidant capacity.

• The Korea Journal of Herbology
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• v.21 no.1
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• pp.101-107
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• 2006

Objectives : It is well known that hexavalent chromium has toxic effect on normal cells. Recently, toxic effect of hexavalent chromium is diminished by the some extracts derived from herbs or plants. But, the toxic or protective mechanism of hexavalent chromium is well unknown. This study was performed to examine the protective effect of poncirin against $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Methods : The protective effect of the cytotoxicity induced by $Na_2Cr_2O_7$ was measured by the cell viability after NIH3T3 fibroblasts were cultured with or without $Na_2Cr_2O_7$ for 48 hours. Antitoxic effects of poncirin on the cytotoxicity induced by $Na_2Cr_2O_7$ were examined by colorimetric assays such as MTT or XTT assay. Results : $Na_2Cr_2O_7$ decreased cell viability by the decreased absorbance in MTT or XTT assay, but, the poncirin increased cell viability which was decreased by $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Conclusion : These results suggest that $Na_2Cr_2O_7$ showed cytotoxicity effect on NIH3T3 fibroblasts by the decrease of cell viavility, and poncirin was effective in the protection of $Na_2Cr_2O_7$-induced cytotoxicity in these cultures.

• Journal of Animal Science and Technology
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• v.66 no.1
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• pp.204-218
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• 2024

Elsholtzia fruticosa (EF) is present in tropical regions throughout South Asian countries as well as the Himalayas. Although it has been used as a traditional medicine to treat digestive, respiratory, and inflammatory issues, its effect on preadipocyte differentiation is unknown. In this study, we examined the effects of a methanol extract prepared from EF on the differentiation of 3T3-L1 preadipocytes. Cell differentiation was assessed by microscopic observation and oil-red O staining. The expression of adipogenic and lipogenic genes, including PPARγ and C/EBPα, was measured by western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR), to provide insight into adipogenesis and lipogenesis mechanisms. The results indicated that EF promotes the differentiation of 3T3-L1 preadipocytes, with elevated lipid accumulation occurring in a concentration-dependent manner without apparent cytotoxicity. EF enhances the expression of adipogenic and lipogenic genes, including PPARγ, FABP4, adiponectin, and FAS, at the mRNA and protein levels. The effect of EF was more pronounced during the early and middle stages of 3T3-L1 cell differentiation. Treatment with EF decreased C/EBP homologous protein (CHOP) mRNA and protein levels, while increasing C/EBPα and PPARγ expression. Treatment with EF resulted in the upregulation of cyclin E and CDK2 gene expression within 24 h, followed by a decrease at 48 h, demonstrating the early-stage impact of EF. A concomitant increase in cyclin-D1 levels was observed compared with untreated cells, indicating that EF modulates lipogenic and adipogenic genes through intricate mechanisms involving CHOP and cell cycle pathways. In summary, EF induces the differentiation of 3T3-L1 preadipocytes by increasing the expression of adipogenic and lipogenic genes, possibly through CHOP and cell cycle-dependent mechanisms.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1576-1583
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• 2006

In order to investigate the effect of Goamshimshinhwan(GASSW) on SD rats with deteriorated immunity caused by methotrexate. Methotrexate was fed to the SD rats once a day for 4 days. After the immune responses of the at a dosage 1,000, 500 and 250mg/kg/10ml. and the changes on body weight and gains, spleen weight, total blood leukocyte numbers, total lymphocyte numbers, the percentage of B-cell, T-cell, CD3+CD+4 T-cell, CD3+CD8+ T-cell and CD4+/CD8+ T-cell ratios in the bolld and spleen were observed. In addition, the serum IL-2 levels and productivity of IL-2 of splenic cells were also demonstrated in this study. The changes on body weight were increased significantly in 100 and 500mg/kg of GASSW groups and the changes on body gain were increased significantly in 1000mg/kg of GASSW groups as compared with control group. The changes on the spleen weight (absolutely or relatively) were increased significantly in all GASSW groups as compared with control group. The total blood leukocyte numbers were increased significantly in 1000 and 500mg/kg of GASSW groups as compared with control group. The total lymphocyte numbers were increased significantly in all GASSW groups in the blood and increased significantly in 1000 and 500mg/kg of GASSW goups in spleen as compared with control group. The percentage of B-cell and T-cell were increased significantly in 1000mg/kg of GASSW groups in the blood and increased significantly in 1000 and 500mg/kg of GASSW groups in spleen as compared with control group. The percentage of CD3+CD4+ T-cell and the serum IL-2 levels and productivity of IL-2 of splenic cells were increased significantly in 100 and 500 mg/kg of GASSW groups in the blood and spleen as compared with control group. The percentage of CD3+CD8+ T-cell were increased significantly in 1000mg/kg of GASSW groups only in spleen as compared with control groups. The CD4+/CD8+ T-cell ratios were increased significantly in 1000 and 500mg/kg of GASSW groups only in the blood as compared with control group. Goamshimshinhwan(GASSW) has immuno-stimulating effect on SD rats with deteriorated immunity caused by methotrexate.

• KSBB Journal
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• v.28 no.6
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• pp.394-399
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• 2013

Astragalus membranaceus Bunge is used in herbal medicine in Eastern Asian countries including Korea. In this study, we assessed the effects of A. membranaceus extract (AM) on the aquaporin-3 (AQP3) protein expression in HaCaT cells. AM did not affect viability of HaCaT cells. AQP3 expression and cell migration seem to be maximal at $100{\mu}g/mL$ concentration. Epidermal growth factor receptor (EGFR) kinase inhibitor, PD153035, blocked AM-induced AQP3 expression and cell migration. In addition, an 80% ethanol extracts of herbal prescription, SinhyoTakleesan (ST), which is composed of A. membranaceus, Angelicae gigantis, Glycyrrhiza glabra Linne, and Lonicera japonica Flos also induced AQP3 expression at $20{\mu}g/mL$ in HaCaT cells. Collectively, these results suggest that AM induce AQP3 expression via EGFR pathway.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.6
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• pp.617-624
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• 2007

Insulin-like growth factor(IGF) is the most abundant growth factor in bone matrix. Recent studies have shown that it can sensitize apoptotic cell death of osteoblasts. Thus, this study investigated whether IGF-II aggravates irradiation-induced cell death of osteoblasts. Cultured MC3T3 osteoblasts were irradiated and IGF-II was added at the concentration of 50 ng/ml immediately after the irradiation. Cell viability was measured by MTT assay. Changes in cell death and cell cycle were analyzed by flow cytometry. The expression of proapoptotic gene bax and antiapoptotic gene bcl-2 was quantified by real time RT-PCR and Western blot. A dose of 30 Gy caused G2/M arrest and increased cell death through both necrosis and apoptosis, while irradiation from 4 to 10 Gy little affected cell cycle and death. IGF-II treatment reduced cell viability without stimulating cell proliferation and changing cell cycle. Combined treatment of IGF-II with irradiation decreased cell viability and proliferation and increased cell death along with G2/M arrest. These effects were not different from those of irradiation only. At transcriptional and protein levels, IGF-II treatment did not affect bax and bcl-2 expression, whereas irradiation increased the expression ofbax without changes in bcl-2. IGF-II in combination with irradiation showed similar findings. These results suggest that IGF-II could modulate apoptotic cell death through mechanisms other than an imbalance between bax and bcl-2 gene expression, although its effect was overridden by irradiation.

• The Korean Journal of Pharmacology
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• v.23 no.1
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• pp.45-49
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• 1987

Employing the approach to isolate the genes expressed preferentially in cytolytic T cell (CTL) but not in other types of cell, 3 CTL-specific cDNAs were recently cloned. To characterize these cDNA clones in relation to CTL activation, their expression pattern after T cell antigen receptor (TCR) or interleukin 2 (IL-2) stimulation were investigated by RNA blot analysis of cloned CTL L3 cells. Transcripts level of two cDNA clones were markedly elevated by TCR stimulation but not by IL-2. In addition, transcripts expression of both clones were abrogated by cyclosporin A treatment. These results indicated that gene activation mediated by TCR is distinct from that mediated by IL-2 and imply that those two unidentified cDNA clones are related to TCR-mediated, IL-2-independent but cyclosporin A-sensitive pathway for CTL activation.