• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.05a
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• pp.183-184
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• 2003

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.

• Journal of Chest Surgery
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• v.36 no.2
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• pp.86-90
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• 2003

Bulla is an air-filled space within the lung parenchyma resulting from deterioration of the alveolar tissue. Molecular mechanism of the formation of the bulla is not well described. Fibroblast growth factor(FGF)-7, bone morphogenetic protein(BMP) receptor, and transforming growth factor(TGF)-$\beta$ receptor are known to have a stimulatory or inhibitory role in the lung formation. We investigated to see if these growth factor or cytokine receptors are involved in the bulla formation by immunohistochemical staining of bullous lung tissues from patients with primary spontaneous pneumothorax. Material and Method: Bullous lung tissues were obtained from 31 patients with primary spontaneous pneumothorax, including 30 males and 1 female from 15 to 39 years old. The bullous tissues were obtained by video-thoracoscopic surgery and/or mini-thoracotomy and fixed in formalin. Blocks of the specimens were embedded with paraffin and cut into 5-6 ${\mu}{\textrm}{m}$ thick slices. The sections were deparaffinized and hydrated and then incubated with primary antibodies against FGF-7, BMP-RII, or TGF-RII. Result: Of the 31 patients, 24 were TGF-RII positive including 18 strong and 6 weak positives. Observation with high magnification showed that strong immunostaining was detected in the boundary region between bullous and normal lung tissues. In contrast, all of the sections were negative with FGF-7 or BMP-RII antibodies. Conclusion: These results suggest that overexpression of TGF- P RII may be involved in the formation of bulla, although further molecular studies are needed to find out more detailed molecular mechanisms.

• Environmental Mutagens and Carcinogens
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• v.24 no.1
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• pp.33-39
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• 2004

To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals including agrochemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. Pyrazosulfuron-ethyl [Ethyl-5-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)-1-methylpyrazole-4-carboxylate, $C_{14}H_{18}N{6}O_{7}S,$ M.W. =414.39, CAS No. 93697-74-6], is one of well known rice herbicide belong in the sulfonyl urea group. To clarify the genotoxicity of this agrochemical, Ames bacterial reversion assay, in vitro chromosomal aberration assay with Chinese hamster lung (CHL) fibroblast and bone marrow micronucleus assay in mice were subjected. In Ames assay, although pyrazosulfuron-ethyl revealed cytotoxic at 5,000-140 $\mug/plate$ in Salmonella typhimurium TA100, no dose-dependent mutagenic potential in 4.4～70 $\mug/plate$ of S. typhimurium TA 98, TA 100, TA1535 and TA 1537 both in the absence and presence of S-9 metabolic activation system was observed. Using CHL fibroblasts, the 50% cell growth inhibition concentration $(IC_{50})$ of pyrazosulfuron-ethyl was determined as 1,243 $\mug/mL,$ and no chromosomal aberration was observed both in the absence and presence of S-9 mixture in the concentration range of 311-1,243 $\mug/mL.$ And also, in vivo micronucleus assay using mouse bone marrow, pyrazosulfuron-ethyl revealed no remarkable induction of MNPCE (micronucleated polychromatic erythrocytes/1000 polychromatic erythrocytes) in the dose range of 625-2,500 mg/kg body weight when administered orally. Consequently, Ames bacterial gene mutation with Salmonella typhimurium, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pyrazosulfuron-ethyl in this study.

• Journal of Food Hygiene and Safety
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• v.21 no.2
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• pp.107-112
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• 2006

Guh Sung Y.L.S-95 (GS95) is a kind of polyacidic solution, which contains acetic acid as a main component. We investigated in the present study tile genetic toxicity of GS95 according to the standard operation procedure from Korean Institute of Toxicology. In the Salmonella typhimurium reverse mutation assay using TA1535, TA1537, TA98 and TA100, GS95 did not induce mutation up to $5,000{\mu}g/plate$. GS95 did not induce chromosome aberration in Chinese hamster lung fibroblast in the concentration range between 1.25 and 5 mg/mL. In the rodent micronucleus assay, the frequency of micronucleated polychromatic erythrocyte in GS95 treated mice were not increased up to 5,000 mg/kg compared to the vehicle treated mice. Taken all these data together, GS95 was proven to be nongenotoxic in the concentration ranges tested.

• Journal of Pharmacopuncture
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• v.13 no.1
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• pp.53-61
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• 2010

Purpose : To investigate the effects of the lavender, lemon and eucalyptus oil mixture on the atopy dermatitis skin lesions induced on NC/Nga Mice by dinitrochlorobenzene (DNCB). Material and Method : For this purpose, we fabricated the oil mixture blending three essential oils (lavender, lemon, eucalyptus : ELL) with one carrier oil (jojoba) and apply it on the atopic dermatitis skin lesions of NC/Nga Mice. Atopic dermatitis in NC/Nga Mice was induced by DNCB treatment on the dorsal skin of mice for 8 weeks. The mixture of ratio of each essential oil drop was 1 (eucalyptus) : 2 (lemon) : 2 (lavender) and this mixture was blended with jojoba oil 50ml (0.025%). The ELL-ointment was supplied for 8 weeks. We evaluated the effects of ELL on cell viability of mouse lung fibroblast, clinical skin features and severity, the level of serum Immunoglobulin (Ig) E & Ig G1, Interleukin (IL)-4, IL-13 and Interferon (IFN)-$\gamma$. Results : ELL showed safety on the cell viability of mouse lung fibroblast compared with control group. The cell viability was measured by SRB method. The effects of ELL on clinical skin features and severity in DNCB-induced dermatitis model of NC/Nga mice was significant compared with control group. EEL also showed significant effects on clinical symptom score compared with control group. Serum IgE & IgG1 level and development of atopy dermatitis skin lesions were evaluated. Serum IgE & IgG1 production was significantly down-regulated in EEL group compared with control group. ELL also down-regulated the levels of IL-4 and IL-13, and up-regulated the level of IFN-$\gamma$ compared with control group significantly. Conclusion : ELL was effective on atopy dermatitis by modulating Th2 related factors.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.4
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• pp.54-71
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• 2005

Purpose : The purpose of this research was to investigate the effects of Saurui Herba Seu Rhizoma(SHSR) on Anti-inflammatory properties in Raw264.7 cell line and murine models of inflammation. Methods : To investigate the effects of Saurui Herba Seu Rhizoma(SHSR) on anti-inflammation, we study cytotoxicity effects of SHSR on Mouse Lung Fibroblast Cells and Peritoneal Macrophages, Inhibitory effects of SHSR on the nitric oxide (NO) release, the ROS production, and the interleukin-6 production. Results : The cytotoxicity of SHSR on mouse lung fibroblast Cells and Raw264.7 cell line was not observed. SHSR in RAW264.7 cell line inhibited $IL-1{\beta}$, IL-6 mRNA gene expression depending upon the concentrations of extract and inhibited IL-18 mRNA gene expression at 100 ${\mu}g/ml$ of extract. SHSR in RAW264.7 cell line inhibit COX-2 mRNA gene expression at 100, 10 ${\mu}g/ml$ of extract. SHSR in RAW264.7 cell line inhibited NOS-II mRNA gene expression depending upon the concentrations of extract. SHSR in RAW264.7 cell line didn't inhibit $TNF-{\alpha}$ mRNA　gene expression. SHSR in RAW264.7 cell line decreased IL-6 production depending　upon the concentrations of extract. SHSR in RAW264.7 cell line decreased $ITNF-{\alpha}$ production according to the concentrations of extract. SHSR in RAW264.7 cell line inhibited NO release specially SHSR 100, 10 ${\mu}g/ml$ concentrations of extract. SHSR inhibit ROS production depending upon the concentrations of extract. Conclusion : These results suggest that SHSR can be used treating a lot of women disease caused by inflammation.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.796-805
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• 1997

Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

• Tuberculosis and Respiratory Diseases
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• v.67 no.6
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• pp.528-535
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• 2009

Background: Dust clouds blown by the wind from the arid deserts of Mongolia and Northeast China are known as Asian dust storms. Ambient particulate matter with a diameter <10 ${\mu}m$ ($PM_{10}$) is associated with the exacerbation of respiratory diseases and increased mortality of heart and lung disease patients. The fibrotic effects of $PM_{10}$ of Asian dust to pulmonary fibroblast cells are unknown. This study examined the production of reactive oxygen species (ROS), TGF-${\beta}$, NF-${\kappa}B$, PDGF-$\alpha$ and Fibronectin in fibroblasts exposed to Asian dust particles. Methods: Air samples were collected using a high volume air sampler (Sibata model HV500F) with an air flow of 500 L/min for at least 6 hours. The MRC-5 cells were exposed to 0, 50 and 100 ${\mu}g/mL$ of $PM_{10}$ for 24 hours. ROS was detected by measuring the level of oxidized DCF using FACS. TGF-$\beta$, NF-${\kappa}B$, PDGF-$\alpha$ and fibronectin were detected by western blotting. Results: There was no increase in the ROS, TGF-$\beta$ and PDGF-$\alpha$ levels in the MRC-5 cells exposed to $PM_{10}$. The NF-${\kappa}B$ level was higher in the MRC-5 cells exposed to 50 and 100 ${\mu}g/mL$ of $PM_{10}$ for 24 hours. The fibronectin level in the MRC-5 cells after 24 hours incubation with 50 ${\mu}g/mL$ $PM_{10}$ was significantly higher than the control group ($PM_{10}$ 50 ${\mu}g/mL$ 113.27${\pm}$8.65 of control, p=0.005). Conclusion: $PM_{10}$ from Asian dust increases the activation of NF-${\kappa}B$ and fibronectin expression in MRC-5 fibroblast cells.

• Tuberculosis and Respiratory Diseases
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• v.71 no.2
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• pp.88-96
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• 2011

Background: It is known that cigarette smoke (CS) causes cell death. Apoptotic cell death is involved in the pathogenesis of CS-related lung diseases. Some members of the protein kinase C (PKC) family have roles in cigarette smoke extract (CSE)-induced apoptosis. This study was conducted to investigate the role of PKC epsilon in CSE-induced apoptosis in human lung fibroblast cell line, MRC-5. Methods: Lactate dehydrogenase release was measured using a cytotoxicity detection kit. The MTT assay was used to measure cell viability. Western immunoblot, Hoechst 33342 staining and flow cytometry were used to demonstrate the effect of $PKC{\varepsilon}$. Caspase-3 and caspase-8 activities were determined using a colorimetric assay. To examine $PKC{\varepsilon}$ activation, Western blotting was performed using both fractions of membrane and cytosol. Results: We showed that CSE activated $PKC{\varepsilon}$ by demonstrating increased expression of $PKC{\varepsilon}$ in the plasma membrane fraction. Pre-treatment of $PKC{\varepsilon}$ peptide inhibitor attenuated CSE-induced apoptotic cell death, as demonstrated by the MTT assay (13.03% of control, 85.66% of CSE-treatment, and 53.73% of $PKC{\varepsilon}$ peptide inhibitor-pre-treatment, respectively), Hoechst 33342 staining, and flow cytometry (85.64% of CSE-treatment, 53.73% of $PKC{\varepsilon}$ peptide inhibitor-pre-treatment). Pre-treatment of $PKC{\varepsilon}$ peptide inhibitor reduced caspase-3 expression and attenuated caspase-3, caspase-8 activity compared with CSE treatment alone. Conclusion: $PKC{\varepsilon}$ seem to have pro-apoptotic function and exerts its function through the extrinsic apoptotic pathway in CSE-exposed MRC-5 cells. This study suggests that $PKC{\varepsilon}$ inhibition may be a therapeutic strategy in CS-related lung disease such as chronic obstructive pulmonary disease.

• Tuberculosis and Respiratory Diseases
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• v.77 no.2
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• pp.73-80
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• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.