A new nucleoside from the Oenothera biennis L / 药学学报

Seven nucleoside compounds were isolated from the Oenothera biennis L. by various chromatographic techniques such as Diaion HP-20, silica gel, Sephadex LH-20, MCI and semi-preparative HPLC. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as 9-(3′-carbonyl methyl)hydroxypurine (1), 1-(3′-carbonyl methyl)purine-6,8-dione (2), N-methyl-2-pyridone-5-carboxamide (3), uracil (4), uridine (5), thymidine (6) and 2′-Ο-methoxy luridine (7). Compound 1 is a new nucleoside and compounds 2-7 were newly isolated from the Oenothera biennis L. Compounds 1-2 can significantly increase the viability of BEAS-2B cells induced by TGF-β1, showing potent anti-pulmonary fibrosis activity.

Evaluation of the viability of BEAS-2B cells exposed to gasoline engine exhaust with different particle sizes by air-liquid interface / 中华劳动卫生职业病杂志

Objective@#To evaluate the viability of gasoline engine exhaust (GEE) with different particle sizes on human lung cell line BEAS-2B in vitro by air-liquid interface (ALI) .@*Methods@#GEE were collected with a Tedlar bag and their particulate matter (PM) number, surface and mass concentration in three kind of GEE (filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter) were measured by two type of particle size spectrometer including TSI-3321 and SMPS-3938. Five groups were included, which divided into blank control group, clean air group, filtered automobile exhaust group, non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group. Except the blank control group, BEAS-2B cells, cultured on the surface of Transwells, were treated with clean air or GEE by ALI method at a flow rate of 25 ml/min, 37 ℃ for 60 min in vitro. CCK-8 cytotoxicity test kit was used to determine the cell relative viability of BEAS-2B cells.@*Results@#In the filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter, high concentrations of fine particles can be detected, but the coarse particles only accounted for a small proportion, and the sequence of PM concentration was motorcycle exhaust without three-way catalytic converter group> non-filtered automobile exhaust group> filtered automobile exhaust group (P<0.001) . Compared with the clean air group, the cell relative viability in the 3 GEE-exposed groups were significantly lower (P<0.001) . Among the comparisons of GEE exposure groups with different particle size spectra, the sequence of the cell relative viability was filtered automobile exhaust group >non-filtered automobile exhaust group> motorcycle exhaust without three-way catalytic converter group (P<0.001) . When took the clean air control group as a reference, the mean of the cell relative viability in the filtered automobile exhaust group, non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group, was decreased by 26.34%, 36.00% and 49.59%, respectively.@*Conclusion@#GEE with different particle size spectra could induce different levels of toxic effects to the human lung cells BEAS-2B by ALI. After lowering the concentration of particles in the GEE and using the three-way catalytic converter could obviously improve the survival rate of lung cells.

2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-glycoside attenuates the apoptotic responses of hypoxia/reoxygenation injury in bronchial epithelial cells through MAPK, HIF-1α and p53 pathway / 药学学报

This study was designed to investigate the effect of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glycoside(TSG)on hypoxia/reoxygenation(H/R)-induced oxidative stress injury and its potential mechanism in human bronchial epithelial cell(BEAS-2B)cells. BEAS-2B cells were exposed to H/R treatment. Level of intracellular ROS was detected using DCFH-DA probe and fluorescence microplate reader. Production of MDA and activity of SOD were evaluated with MDA and SOD kits. Nucleus was shaped by DAPI staining. Translocation of Bax to mitochondria was observed in MCF-7/GFP-Bax cells. Change in mitochondrial membrane potential was detected by JC-1 staining. Release of cytochrome C from mitochondria was detected by immunofluorescence. Expressions of mitochondrial/cytoplasmic Bax and cytochrome C, caspase-9, caspase-3, phosphorylated MAPK, HIF-1α and phosphorylated p53(p-p53)were determined by Western blotting. TSG significantly improved cell viability and reduced H/R-induced ROS production in BEAS-2B cells, while significantly decreased MDA production. It inhibited Bax translocation and nucleus fracture, reversed the decrease in mitochondrial membrane potential and inhibited the release of cytochrome C and following activation of caspase-9/caspase-3. Simultaneously, TSG down-regulated the signals of SAPK JNK1/2 and p38 MAPK without an impact in ERK1/2. It attenuated expression of HIF-1α and phosphorylation of p53. This study suggests that TSG could protect BEAS-2B against H/R-induced apoptosis, perhaps through the MAPK, HIF-1α and p53 pathways.

Oxidative damage of BEAS-2B cells induced by depleted uranium and protection by DMSO / 中华放射医学与防护杂志

Objective To observe the oxidative damage in human bronchial epithelial cells(BEAS-2B) induced by depleted uranium(DU)and protection of DMSO.Methods The measurement of extracellular superoxide anions(O2-·)was based on the reduction of ferricytochrome C.Quantitative analysis of extracellular hydrogen peroxides(H2O2)was used by the horseradish peroxidase-dependent oxidation of phenol red.The determination of extracellular hydroxyl radicals(·OH)was based on discoloration of safranine T.Ethidium bromide and 2,7'-dichlorofluorescein,fluorescent products of the membrane-permeable dyes-hydroethineand 2,7'-dichloroflurescin diacetate were used to monitor the intracellular production of O2-·and H2O2 by fluorometric method.The enzyme activity of SOD and GSH were measured by chemiluminescence and spectrophotometric method,respectively.Results The ROS production,including H2O2,O2-·and·OH,increased remarkably which induced by DU in BEAs-2B cells.The enzyme activity of SOD and GSH was descended remarkedly.These changes could be effectively inhibited by 0.5% of DMSO.Conclusions DU causes oxidative damage to BEAS-2B cells.Through removing active oxygen,DMSO can inhibit oxidative damage of DU.

Reconstruction of Tissue Engineered 3D Bronchial Model in Vitro / 中国康复理论与实践

@#Objective To reconstruct tissue-engineered 3D bronchial model using human bronchial epithelial cells and human embryo lung fibroblast as seeding cells, and liquid collagen mixed Matrigel as scaffold. Methods Human bronchial epithelial cells and human embryo lung fibroblast were mixed with liquid collagen supplementing with matrigel and casted in 12-wells plate to reconstruct cells-collagen sheet. Macroscopic observation, phase-contrast microscopy observation, routine HE staining and immunohistochemistry staining(CK ets) were employed to assess the engineered 3D model. Results We reconstructed engineered 3D bronchial model successfully in vitro by tissue engineering techniques and exerted static stretch onto the collagen sheet. From Macroscopic observation, we gained contracted well sheet. We also observed network structure in phase-contrast microscopy meanwhile the viability of cells was fine. HE staining showed the formation of 3D network structure. The immunohistochemistry staining of CK and Vimentin were positive.Conclusion We reconstructed engineered 3D bronchial model successfully in vitro and seeding cells could implement polarity growing in the scaffold materials then gained the network structure.

Effects of Nicotine, Cotinine and Benzopyrene as Smoke Components on the Expression of Antioxidants in Human Bronchial Epithelial Cells / 결핵및호흡기질환

BACKGROUND: Cigarette smoking is an important risk factor for chronic bronchitis and COPD. Airway epithelial cells exposed to cigarette smoke components such as nicotine, cotinine and benzopyrene can generate reactive oxygen species (ROS) and be subject to oxidative stress. This oxidative stress can induce the inflammatory response in the lung by the oxidant itself or by the release of proinflammatory cytokines. It has been reported that nicotine stimulates ROS, which are associated with NF-kappaB. METHODS: Beas2B cells were treated with nicotine, cotinine and benzopyrene. RT PCR was used to measure the expression of several antioxidant factors using the total RNA from the Beas2B cells. The level of superoxide dismutase(CuZnSOD), thioredoxin, glutathione reductase expression was examined. RESULTS: 0.5 to 4 hours after the benzopyrene, nicotine and cotinine theatments, the level of thioredoxin and glutathione reductase expression decreased. Longer exposure to these compounds for 24 to 72 hours inhibited the expression of most of these antioxidant factors. CONCLUSION: During exposure to smoke compounds, thioredoxin and glutathione reductase are the key antioxidant factors induced sensitively between 0.5 and 4 hours but the levels these antioxidants decrease between 24 hour and 72hours.

The effect of respiratory syncytial virus infection on neutrophil adherence to airway epithelial cells / 천식및알레르기

BACKGROUND: Respiratory syncytial virus (RSV) is the most common cause of lower respiratory infection in infants and young children, but the pathogenesis of RSV-induced inflammation is not well defined. MATERIAL AND METHOD: In order to examine the potential interactions between virus-infected airway epithelial cells and neutrophils, we studied the ability of neutrophils to adhere to yirus-infected airway epithelial cell monolayers by myeloperoxidase assay. Also we measured the ability of airway epithelial cells to secrete interleukin-8(IL-8) and inter-cellular adhesion molecule-1(ICAM-1) in virus-infected airway epithelial cell cultures by enzyme-linked immunosorbent assay(ELISA). The degree of IL-8 and ICAM-1 gene expression in the RSV-infected BEAS-2B cell cultures were analyzed by reverse transcription-polymerase chain reaction(RT-PCR). RESULTS: The RSV-infected BEAS-2B cell resulted in significantly enhanced level of neutrophil adherence compared to the uninfected control(p (0.001). IL-8 and ICAM-1 production significantly increased by RSV infection(p<0.05). There was a significant positive correlation between neutrophil adherence and IL-8 level(r=0.73, p=0.002), and ICAM-1 level (r=0.843, p=0.001) in RSV-infected cells. The degree of both IL-8 and ICAM-1 mRNA expression increased in the RSV-infected cells compared with the uninfected ones. CONCLUSION: These results suggest that RSV infection significantly enhances the production of IL-8 and ICAM-1 in airway epithelial cells which then results in increased neutrophil adherence.

Antagonism of DMSO against Genotoxicity of Cooking Oil Fume Condensate to BEAS-2B Cell / 环境与健康杂志

Objective To study the antagonism of DMSO against the toxicity of cooking oil fume condensation to BEAS-2B cell.Methods The comet assay,micronucleus test and multinucleated cells test were used to research the genotoxicity induced by cooking oil fume condensation(COFC)and the antagonism of DMSO.Results COFC induced DNA broken,the tail area,rate of comet occurrence,tail length,tail moment,olive tailmoment increased significantly,the frequencies of micronucleus and multinucleated cells were significantly increased and the damage of cells could be inhibited effectively by DMSO.Conclusion The antagonistic effects of DMSO on the toxicity of COFC was significant in BEAS-2B cell.