1.1,25-dihydroxyvitamin D3 pretreatment inhibits house dust mite-induced thymic stromal lymphopoietin release by human airway epithelial cells.
Liqin ZHOU ; Hangming DONG ; Haijin ZHAO ; Mengchen ZOU ; Lihong YAO ; Fei ZOU ; Shaoxi CAI
Journal of Southern Medical University 2014;34(4):492-496
OBJECTIVETo investigate the effect of 1,25-dihydroxyvitamin D3 (1,25VD3) on house dust mites (HDM)-induced expression of thymic stromal lymphopoietin (TSLP) in human airway epithelial cells in vitro.
METHODSHuman airway epithelial 16HBE cells were incubated with 200, 400, and 800 U/L in the absence or presence of 1,25VD3 (10(-8) mol/L) for 6 h and 24 h, and TSLP mRNA and protein expressions in the cells were assessed using quantitative PCR and ELISA.
RESULTS16HBE cells incubated with HDM at 200, 400, and 800 U/L showed significantly increased TSLP mRNA and protein expressions (P<0.05). Pretreatment of the cells with 1,25VD3 obviously lowered 400 U/L HDM-induced TSLP expressions (P<0.05), but 1,25VD3 added along with HDM in the cells did not produce significant effects on TSLP expressions (P=0.58).
CONCLUSIONBoth 1,25VD3 and HDM can induce TSLP expression and release in 16HBE cells, but pretreatment with 1,25VD3 can decrease HDM-augmented TSLP expression in the cells.
Animals ; Bronchi ; cytology ; Calcitriol ; pharmacology ; Cell Line ; Cytokines ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Pyroglyphidae
2.Dexamethasone decreases IL-29 expression in house dust mite-stimulated human bronchial epithelial cells.
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(6):823-827
The aim of this study was to explore the effect of IL-29 on the progression of airway allergic disease by detecting the level of IL-29 in airway allergic cell models stimulated by house dust mite (HDM) in the presence or absence of dexamethasone (DEX). The same batch of human bronchial epithelial cells in exponential growth phase was randomly divided into five groups: blank group (A), 300 ng/mL HDM group (B), 1000 ng/mL HDM group (C), 3000 ng/mL HDM group (D), and 300 ng/mL HDM+100 ng/mL DEX group (E). The IL-29 mRNA expression was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The IL-29 protein expression in cell suspension was detected by ELISA. The results showed that after stimulation with HDM for 24 h, the expression of IL-29 was increased significantly, and after co-stimulation with HDM and DEX for 24 h, the expression of IL-29 in group E was significantly lower than that in the groups stimulated by HDM alone but higher than that in the group A. The differences between the different groups were significant (F=132.957, P<0.01). Additionally, the higher the concentration of HDM was, the more significant the increase in the IL-29 expression was. In conclusion, IL-29 may play a role in the progression of airway allergic disease including asthma.
Adult
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Animals
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Bronchi
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cytology
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drug effects
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metabolism
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Cells, Cultured
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Dexamethasone
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pharmacology
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Epithelial Cells
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drug effects
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metabolism
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Humans
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Interleukins
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metabolism
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Mites
3.Hydrogen peroxide induces high mobility group box 1 release in human bronchial epithelial cells.
Changchun HOU ; Haijin ZHAO ; Wenjun LI ; Shaoxi CAI
Journal of Southern Medical University 2012;32(8):1131-1134
OBJECTIVETo investigate the effect of hydrogen dioxide (H(2)O(2)) on the release and translocation of high mobility group box 1 release (HMGB1) from normal human bronchiolar epithelial cells (HBE).
METHODSMTT assay was used to assess the viability of HBE135-E6E7 cells exposed to different concentrations of H(2)O(2). The expression and location of HMGB1 in the cytoplasm, nuclei and culture medium of the exposed cells were determined using Western blotting and immunofluorescence assay.
RESULTSExposure to 125 µmmol/L H(2)O(2) did not obviously affect the cell viability. At the concentration of 250 µmmol/L, H(2)O(2) significantly decreased the cell viability (P<0.05), but significant cell death occurred only after exposure to 400 µmmol/L H(2)O(2) (P=0.000). Compared with the control cells, the cells exposed to 12.5, 125 and 250 µmmol/L H(2)O(2) for 24 h showed significantly increased levels of HMGB1 in the culture medium (P<0.05), and exposure to 125 µmmol/L H(2)O(2) for 12 and 24 h also caused significantly increased HMGB1 level (P<0.05). Exposure to 125 µmmol/L H(2)O(2) for 24 h significantly increased HMGB1 expression in the cytoplasm but decreased its expression in the nucleus. HMGB1 translocation from the nuclei to the cytoplasm and to the plasmalemma occurred after 125 µmmol/L H(2)O(2) exposure for 12 h and 24 h, respectively.
CONCLUSIONH(2)O(2) can induce HMGB1 translocation and release in human bronchial epithelial cells, suggesting the involvement of HMGB1 in airway oxidative stress in chronic inflammatory diseases such as asthma and COPD.
Bronchi ; cytology ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; HMGB1 Protein ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Protein Transport
4.Effect of thymic stromal lymphopoietin on human bronchial epithelial permeability.
Wenjia LI ; Haijin ZHAO ; Hangming DONG ; Fei ZOU ; Shaoxi CAI
Journal of Southern Medical University 2014;34(6):802-806
OBJECTIVETo investigate the effect of thymic stromal lymphopoietin (TSLP) on the permeablily of monolayer bronchial epithelial cells in vitro.
METHODSCultured human bronchial epithelial cell line 16HBE was exposed to 0.1 or 1 ng/ml TSLP for 0, 0.5, 6, 12, or 24 h, and the epithelial monolayer permeability was assessed by measuring transepithelial electrical resistance (TER), permeability to FITC-labeled dextran (FITC-DX) and expression of E-cadherin.
RESULTSCompared with the control cells group, 16HBE cell monolayer showed significantly increased TER (P<0.001) and decreased FITC-DX fluorescence in the lower chamber (P<0.05) following exposure to 0.1 and 1 ng/ml TSLP, but these changes were not dose-dependent. Exposure to 0.1 ng/ml TSLP resulted in significantly increased expression of E-cadherin. The 16HBE monolayer exposed to 0.1 ng/ml TSLP for 24 h showed the most obvious increase of TER and E-cadherin expression (P<0.05); FITC-DX fluorescence level was markedly decreased after TSLP exposure for 12 h and 24 h (P<0.05), and the effect was more obvious in 12 h group.
CONCLUSIONTSLP can protect the barrier function of normal bronchial epithelial cells in vitro.
Bronchi ; cytology ; Cadherins ; metabolism ; Cell Line ; Cytokines ; pharmacology ; Epithelial Cells ; drug effects ; Humans ; Permeability
5.Effect of miR-542-3p on carcinogenesis induced by anti-benzo(a) pyrene-7,8-diol-9,10-epoxide.
Yao ZHAO ; Huan-ying LIU ; Yuan-qi LI ; Yi-guo JIANG
Chinese Journal of Preventive Medicine 2011;45(5):416-421
OBJECTIVETo explore the effect of miR-542-3p in malignant transformation of human bronchial epithelial cells (16HBE) induced by anti-benzo(a)pyrene-7,8-diol-9,10-epoxide (anti-BPDE).
METHODSThe relative expression level of mature miR-542-3p in transformed cells (16HBE-T) and untransformed control cells (16HBE-N) was measured by real-time quantitative polymerase chain reaction (qRT-PCR). miRNA mimic was transiently transfected into 16HBE-T to change the expression level of miR-542-3p, and then the influenced changes of cell proliferation, cell cycle, apoptosis, and soft agar colony formation rate and the migration of transfected cells were analyzed.
RESULTSBefore transfection, the expression level of mature miR-542-3p in 16HBE-T was lower (39.08 ± 6.95)% than it in 16HBE-N (t = 15.18, P < 0.05). In comparison with the 16HBE-T group, the expression level of miR-542-3p in miR-542-3p mimic-transfected group was (5.23 ± 0.55) fold (t = 17.37, P < 0.05) after transfection. Cell proliferation of mimic-transfected group was decreased to (62.06 ± 5.61)% (t = -17.28, P < 0.05), percentage of cells in G(0)/G(1) phase up to (74.76 ± 4.86)% (t = 4.53, P < 0.05), rate of colony formation degrade to (5.87 ± 0.67)% (t = -6.66, P < 0.05), coverage areas ratio decreased to (0.31 ± 0.08) (t = -6.78, P < 0.05). There was no change with apoptosis.
CONCLUSIONOur studies showed that miR-542-3p played the role as a tumor suppressor, which led to a significant decrease in the proliferation capacity and degree of malignancy. These findings suggest aberrantly down-regulated miR-542-3p may be one critical factor that contributes to malignant transformation of 16HBE induced by anti-BPDE.
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; adverse effects ; Bronchi ; cytology ; Cell Transformation, Neoplastic ; drug effects ; genetics ; metabolism ; Epithelial Cells ; cytology ; drug effects ; Humans ; MicroRNAs ; genetics ; Transfection
6.The effects of the erythromycin on the production of r-glutamylcys glutamylcysteine synthetase and glutathione in the bronchial epithelial cell.
Iiang YU ; Bing LI ; Pixin RAN
Chinese Journal of Applied Physiology 2009;25(1):101-132
Bronchi
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cytology
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metabolism
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Cell Line
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Epithelial Cells
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cytology
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metabolism
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Erythromycin
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pharmacology
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Glutamate-Cysteine Ligase
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genetics
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metabolism
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Glutathione
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genetics
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metabolism
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Humans
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RNA, Messenger
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genetics
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metabolism
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Up-Regulation
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drug effects
7.25-hydroxyvitamin D3-induced increases of normal human airway epithelial cell permeability is not mediated by upregulated ZO-1 expression.
Xiao-ting ZHOU ; Hai-jin ZHAO ; Shao-xi CAI
Journal of Southern Medical University 2011;31(7):1187-1189
OBJECTIVETo observe the effect of 25-hydroxyvitamin D3 on the permeability and ZO-1 expression in normal human airway epithelial cells.
METHODSMTT assay was used to assess the viability of human airway epithelial cell line 16HBE following a 24-hour exposure to different concentrations of 25-hydroxy vitamin D3, and the transepithelial electrical resistance (TER) of the cell monolayer was measured using a Millicell-ERS voltohmmeter. Real-time quantitative RT-PCR was employed to determine the changes of ZO-1 mRNA expression in the cells following the exposures.
RESULTSExposure to 25-hydroxyvitamin D3 resulted in significantly increased permeability of 16HBE cells, but the exspression of ZO-1 showed no obvious changes. 25-hydroxyvitamin D3 at 4×10(-9) mol/L showed the strongest effect in increasing the permeability of cell monolayer.
CONCLUSION25-hydroxyvitamin D3 increases the permeability of normal bronchial airway epithelial cell monolayer in vitro, but this effect is not mediated by upregulation of ZO-1 expression.
Bronchi ; cytology ; metabolism ; Calcifediol ; pharmacokinetics ; pharmacology ; Cell Line ; Cell Membrane Permeability ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Humans ; RNA, Messenger ; genetics ; metabolism ; Zonula Occludens-1 Protein ; genetics ; metabolism
8.Identification of protein peroxiredoxin 2 related to crystalline NiS-induced neoplastic transformation.
Wei-dong JI ; Jia-kun CHEN ; Jia-chun LÜ ; Zhong-liang WU ; Su-mei FENG ; Fei YI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(4):267-270
OBJECTIVETo provide evidence for illustrating the molecular mechanism of nickel carcinogenesis, and to identify the differential expression of protein in crystalline NiS-induced neoplastic transformation of human bronchial epithelial cell by proteomics technology.
METHODSTwo dimensional electrophoresis (2-DE) and the ImageMaster 3.10 software were used to analyze the differential expression of protein, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) combined with database search was applied to identify protein peroxiredoxin 2 (PDX2) related to malignant transformation.
RESULTSThe good 2-DE pattern including resolution and reproducibility was obtained. Nearly 700 expressed proteins per 2-D gel were isolated with molecular weights (MW) ranging from 14,400 to 94,000 KD and pI 3 - 10. A protein PDX2 with MW 21,890 KD, pI 5.66, which was highly expressed in malignantly transformed cell, was identified using MALDI-TOF-MS.
CONCLUSIONPDX2 was involved in malignant transformation of human bronchial epithelial cell induced by crystalline nickel sulfide.
Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Nickel ; toxicity ; Peroxiredoxins ; metabolism ; Proteome
9.High-mobility group box protein 1 in synergy with interleukin-1β promotes interleukin-8 expression in human airway epithelial cells in vitro.
Dandan ZHANG ; Haijin ZHAO ; Liqin ZHOU ; Jiafu SONG ; Hangming DONG ; Fei ZOU ; Shaoxi CAI
Journal of Southern Medical University 2012;32(12):1764-1767
OBJECTIVETo test the effect of high-mobility group box protein 1 (HMGB1) alone or in synergy with interleukin-1β (IL-1β) on the expression of IL-8 in human airway epithelial cells in vitro.
METHODSHuman airway epithelial 16HBE and A549 cell lines were incubated with HMGB1 (100 ng/ml) in the absence or presence of IL-1β (10 ng/ml) for 24 h, and the changes of IL-8 mRNA and protein expressions were assessed using quantitative PCR and enzyme-linked immunosorbent assay (ELISA).
RESULTSIn the two human airway epithelial cell lines, HMGB1 alone did not produce obvious effect on the expression of IL-8, but in the presence of IL-1β, HMGB1 caused a significant increase of IL-8 expressions at both the mRNA and protein levels.
CONCLUSIONHMGB1 in synergy with IL-1β increases the expression of IL-8 in human airway epithelial cells, which provides new evidence that HMGB1 contributes to neutrophilic airway inflammation by regulating IL-8 expression.
Bronchi ; cytology ; drug effects ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; HMGB1 Protein ; pharmacology ; Humans ; Inflammation ; Interleukin-1beta ; pharmacology ; Interleukin-8 ; metabolism ; RNA, Messenger
10.Monocrotaline-induced pulmonary hypertension correlates with upregulation of connective tissue growth factor expression in the lung.
Young Sam LEE ; Jonghoe BYUN ; Jeong A KIM ; Jung Sun LEE ; Koung Li KIM ; Yeon Lim SUH ; Jeong Min KIM ; Hyung Suk JANG ; Jae Young LEE ; In Soon SHIN ; Wonhee SUH ; Eun Seok JEON ; Duk Kyung KIM
Experimental & Molecular Medicine 2005;37(1):27-35
Pulmonary hypertension (PH) is characterized by structural and functional changes in the lung including proliferation of vascular smooth muscle cells (VSMCs) and excessive collagen synthesis. Although connective tissue growth factor (CTGF) is known to promote cell proliferation, migration, adhesion, and extracellular matrix production in various tissues, studies on the role of CTGF in pulmonary hypertension have been limited. Here, we examined CTGF expression in the lung tissues of male Sprague Dawley rats treated with monocrotaline (MCT, 60 microgram/kg), a pneumotoxic agent known to induce PH in animals. Establishment of PH was verified by the significantly increased right ventricular systolic pressure and right ventricle/left ventricle weight ratio in the MCT-treated rats. Histological examination of the lung revealed profound muscular hypertrophy in the media of pulmonary artery and arterioles in MCT-treated group. Lung parenchyma, vein, and bronchiole did not appear to be affected. RT-PCR analysis of the lung tissue at 5 weeks indicated significantly increased expression of CTGF in the MCT-treated group. In situ hybridization studies also confirmed abundant CTGF mRNA expression in VSMCs of the arteries and arterioles, clustered pneumocytes, and infiltrated macrophages. Interestingly, CTGF mRNA was not detected in VSMCs of vein or bronchiole. In saline-injected control, basal expression of CTGF was seen in bronchial epithelial cells, alveolar lining cells, and endothelial cells. Taken together, our results suggest that CTGF upregulation in arterial VSMC of the lung might be important in the pathogenesis of pulmonary hypertension. Antagonizing the role of CTGF could thus be one of the potential approaches for the treatment of PH.
Animals
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Blood Pressure/drug effects
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Bronchi/cytology/drug effects/metabolism
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Endothelial Cells/cytology/drug effects/metabolism
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Epithelial Cells/cytology/drug effects/metabolism
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Hypertension, Pulmonary/chemically induced/*metabolism
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Immediate-Early Proteins/genetics/*metabolism
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Intercellular Signaling Peptides and Proteins/genetics/*metabolism
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Lung/cytology/drug effects/*metabolism
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Male
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Monocrotaline/*toxicity
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Pulmonary Alveoli/cytology/drug effects/metabolism
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Pulmonary Artery/cytology/drug effects/metabolism
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Rats
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Rats, Sprague-Dawley
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Research Support, Non-U.S. Gov't
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Reverse Transcriptase Polymerase Chain Reaction
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Up-Regulation