OBJECTIVETo investigate SiO2-induced EMT in human bronchial epithelial cells HBE in vitro.

METHODSHBE cells were cultured and then stimulated with indicated doses of SiO2 (0, 50, 100, 200, 300 µg/ml). The morphological changes were observed by microscope. In addition, Western blot was per-formed to detect the expression of E-cad, α-SMA and Vim. The changes of migration ability were examined by wound-healing assay in vitro.

RESULTS(1) After exposure to SiO2, HBE cells lost contact with their neighbor and displayed a spindle-shape, fibroblast-like morphology. (2) Compared with the control, the E-cad (300 µg/ml group) expression downregulated 2.98 fold (P < 0.05), and the Vim (300 µg/ml group) and α-SMA (200 µg/ml group) expression upregulated 4.46 fold and 3.55 fold (P < 0.05). There were significant differences between 100, 200, 300 µg/ml groups and the control group (P < 0.05). (3) In the test group, the percentage of wound-healing areas/wound areas were larger than those in control group (P < 0.05).

CONCLUSIONSSiO2 could induce EMT in human bronchial epithelial cells.

Bronchi ; cytology ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Silicon Dioxide ; adverse effects ; Stromal Cells ; cytology ; drug effects

OBJECTIVETo investigate the effects of ephedrine on human nasal cilia movement.

METHODSCiliary beat frequency (CBF) of cultured human nasal epithelial cells was measured by high-speed digital microscopy in HBSS and ephedrine solution of different concentrations in 10 minutes.

RESULTSCBF of cultured nasal epithelial cells exposed to HBSS showed no significant changes in 10 minutes. However, in 2.5 g/L , 5 g/L, 10 g/L and 20 g/L ephedrine solution, CBF increased significantly in 1-2 minutes and reached the apex, then it decreased gradually, at the 10th minute. CBF of the samples exposed to 2.5 g/L and 5 g/L ephedrine solution were slower than those in HBSS, but no significant changes were found. However, in 10 g/L and 20 g/L ephedrine solution, CBF decreased significantly when compared with samples in sHBSS. With the concentrations from 2.5 g/L to 20 g/L ephedrine, the increment was independent on the concentration, the inhibitory effect was dependent on the concentration.

CONCLUSIONSIn initial time, 2. 5 g/L-20 g/L ephedrine stimulated CBF, then 10 g/L-20 g/L ephedrine inhibited CBF. The stimulation of 2.5 g/L and 5 g/L ephedrine on CBF was longer than that of 10 g/L and 20 g/L ephedrine. 5 g/L ephedrine had maximum stimulatory effect without obvious inhibitory effect on cultured human nasal CBF.

Cells, Cultured ; Cilia ; drug effects ; physiology ; Ephedrine ; pharmacology ; Epithelial Cells ; drug effects ; physiology ; Humans ; Nasal Mucosa ; cytology ; drug effects ; physiology

OBJECTIVE: To study the effect of erythromycin on apoptosis in epithelial cell and investigate the significance of epithelial cell apoptosis in nasal polyps forming. METHOD: Epithelial cell collected from thirty nasal polyps and six inferior turbinates were cultured in Dulbecco Eagle and Ham F12 (1:1) and divided into two groups, one cultured with Erythromycin(Erythromycin group), another cultured without Erythromycin (control group). Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. RESULT: The AI (apoptosis index) of epithelial cell in nasal polyps after cultured for 1,3,5 days with erythromycin were respectively (33.23 +/- 6.50)%, (38.21 +/- 7.22)% and (52.63 +/- 7.86)%. The AI of epithelial cell in inferior turbinates were respectively (31.02 +/- 5.60)%, (32.13 +/- 7.15)% and (39.64 +/- 7.48)%. There were significant difference between two groups at 5 day after culture (P < 0.05). CONCLUSION Erythromycin promoted apoptosis of epithelial cell in nasal polyps.
Apoptosis ; drug effects ; Epithelial Cells ; drug effects ; Erythromycin ; pharmacology ; Humans ; Nasal Polyps ; pathology

Bronchi ; drug effects ; pathology ; Cell Line ; Cell Transformation, Neoplastic ; drug effects ; Epithelial Cells ; drug effects ; pathology ; Humans ; Mineral Fibers ; toxicity

Animals ; Epithelial Cells ; cytology ; drug effects ; Humans ; Kidney Tubules ; cytology ; drug effects ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Podocytes ; drug effects

OBJECTIVETo detect the expression changes of the demethylase TETs (Ten-eleven translocation enzymes) in human embryonic kidney cell (HEK293) exposed to high dose cadmium chloride (CdCl2), and to investigate the regulation effects of TETs on global genomic methylation.

METHODSHEK293 cells were exposed to CdCl2 for 24 h, 48 h and 72 h, the survival rate was tested by CCK-8 (cell counting kit-8) method, and the cell morphology was observed. The levels of TETs mRNA and protein were detected by fluorescence quantitative PCR and Western blot, respectively. The genomic DNA methylation level was detectedby pyro sequencing assay.

RESULTSCdCl2 had toxic effects on HEK293 cells, and the half inhibitory concentration (IC50) was 1.78 µmol/L. After exposure of CdCl2 for 24 h, 48 h and 72 h, the morphology of HEK293 cells was altered, and the high dose group (2.0 µmol/L) showed vacuolar changes and fuzzy appearance. The level of TET1 mRNA in groups of 0.0, 0.5, 1.0, 2.0 µmol/L were 0.23 ± 0.13, 0.48 ± 0.12, 0.59 ± 0.16 and 0.95 ± 0.39, respectively (F = 182.89, P = 0.002); The level of TET2 mRNA in groups of 0.0, 0.5, 1.0, 2.0 µmol/L were 0.23 ± 0.12, 0.32 ± 0.02,0.31 ± 0.10 and 0.34 ± 0.07, respectively (F = 27.94, P < 0.001); The level of TET3 mRNA in groups of 0.0, 0.5, 1.0, 2.0 µmol/L were 0.26 ± 0.10, 0.27 ± 0.11, 0.25 ± 0.11 and 0.28 ± 0.09, respectively (F = 1.76, P = 0.036). The interaction effect existed between exposure time and doses of TET1 mRNA, TET2 mRNA and TET3 mRNA (F values were 32.94, 23.04 and 13.78, respectively; P values were < 0.001, 0.041 and < 0.001, respectively). Western blot showed that in different exposure time and dose, the protein expression levels of TETs had the similar trend as mRNA levels. In 24 h (55.01 ± 3.62)%, 48 h (48.31 ± 8.99)%, 72 h (48.76 ± 6.60)%, the DNA methylation had significant differences (F = 18.50, P < 0.001); In groups of 0.0 µmol/L (55.29 ± 2.83)%, 0.5 µmol/L (55.35 ± 3.11)%, 1.0 µmol/L (48.58 ± 6.40)% and 2.0 µmol/L (43.56 ± 7.89)%, the differences of DNA methylation had significant differences (F = 7.03, P = 0.048); the effect of interaction was also existed (F = 2.73, P = 0.043).

CONCLUSIONIn the short term exposure to CdCl2, the levels of TETs mRNA and protein showed a trend of increase according to the exposure time and dose, and the methylation level of whole genomic DNA was also altered. The demethylase TETs may play a role in regulating the genomic methylation level of HEK293 exposed to cadmium.

Cadmium Chloride ; toxicity ; DNA Methylation ; Dioxygenases ; genetics ; Epithelial Cells ; drug effects ; HEK293 Cells ; Humans ; RNA, Messenger

This paper aimed to study the protective effect of ginsenoside Rg_1 on endotoxin(LPS)-induced apoptosis of lung epithelial cells and its mechanism of action. Mouse lung epithelial cells(MLE-12) were first treated with LPS. The autophagy changes and apoptosis and the relationship with concentration and time of LPS were observed. Then,the level of autophagy in MLE-12 was regulated at a specific concentration and action time of LPS,and the changes of apoptosis were observed. Secondly,ginsenoside Rg_1 and autophagy inhibitor 3-MA were added respectively at the same concentration and action time of LPS. The lung epithelial cells were grouped to observe the effect of ginsenoside Rg_1 on LPS-induced apoptosis of lung epithelial cells and its mechanism. In the animal experiment,the mice were grouped and tested by apoptosis protein,lung injury score and HE staining section to verify whether ginsenoside Rg_1 has a protective effect on LPS-induced lung injury. The results showed that apoptosis and autophagy increased as the rise of concentration after treatment with LPS for 12 h. The apoptosis increased gradually,and the autophagy increased first and then decreased over time at the LPS concentration of 25 g·L-1. The apoptosis of LPS group was higher than that of control group,and LPS+3-MA group increased further,while apoptosis decreased significantly in LPS+RAM(rapamycin,autophagy promoter) group. The autophagy increased in LPS group,decreased in LPS+3-MA group and increased in LPS+RAM group. The apoptosis of LPS group was higher than that of control group,and the apoptosis of LPS+Rg_1 group decreased. The apoptosis of LPS+Rg_1+3-MA group increased again. The autophagy of LPS group further increased after administration of ginsenoside Rg_1,but decreased after administration of 3-MA. In the in vivo experiments in mice,the apoptosis of LPS group increased significantly compared with the control group,while LPS + ginsenoside Rg_1 group decreased. Lung injury score and HE staining also conformed to the above trend. LPS can induce the apoptosis of lung epithelial cells in a time-dependent and concentration-dependent manner. The autophagy of lung epithelial cells increases with the rise of LPS concentration. At the specific concentration of LPS,autophagy increases first and then decreases after 12-16 hours. Proper increase of autophagy in lung epithelial cells within a certain period of time can reduce the apoptosis induced by LPS,while inhibition of autophagy can increase apoptosis. Ginsenoside Rg_1 has a protective effect on lung cancer epithelial cell apoptosis induced by autophagy.
Animals ; Apoptosis ; Autophagy ; Cells, Cultured ; Epithelial Cells ; drug effects ; Ginsenosides ; pharmacology ; Lipopolysaccharides ; Lung ; cytology ; Mice

OBJECTIVETo explore the effect of silica dioxide(SiO2) on proliferation and downregulation of gap junctional intercellular communication (GJIC) in pulmonary alveolar epithelial cells (CCL-64 cells).

METHODSThe pulmonary alveolar macrophages(PAMs) were incubated in the serum-free RPMI 1640 containing the various concentration of SiO2 for 24 hours. The supernatants were prepared and added 5% (V/V) into 2% (V/V) NBS RPMI 1640 to stimulate the proliferation of CCL-64 cells for 24 hours. A set of "blank control", run in parallel, contained RPMI 1640 + 2% (V/V) NBS alone. The proliferation of CCL-64 cells was detected using MTT assay(to show as the absorbency, A570nm). GJIC function was measured using the fluorescence redistribution after photobleaching(FRAP) assay [to express as the transfer rate of the fluorescence, K (x 10(-3)/s)], with a laser scanning confocal microscope(LSCM, Leica TCS SP).

RESULTSThe silica-exposed PAM supernatants could induce both the proliferation(F = 9.679, P < 0.01) and downregulation of GJIC(F = 20.587, P < 0.01) of CCL-64 cells. In the range of 50-500 micrograms/ml SiO2 concentrations, the proliferation (A570nm values) and GJIC(the transfer rate, K) were fitted well in a dose-dependent manner(proliferation: r = 0.891, P < 0.05; GJIC: r = -0.943, P < 0.05).

CONCLUSIONBy way of stimulating the PAM, SiO2 could inhibit GJIC function in lung alveolar epithelial cells, and induce epithelial cell proliferation. In the pathogenesis of silicosis, the downregulation of GJIC of the pulmonary epithelial cells may play an important role in silica-mediated alveolar epithelial cell injury.

Cell Communication ; drug effects ; Cell Proliferation ; drug effects ; Epithelial Cells ; drug effects ; Gap Junctions ; drug effects ; Pulmonary Alveoli ; drug effects ; Silicon Dioxide ; toxicity ; Silicosis ; etiology

OBJECTIVETo investigate the effect of human serum albumin (HSA) on cell attachment of human gingival epithelial cells (HGE).

METHODSHGE were primary cultured with keratinocyte serum-free medium (KSFM) and dispase. The cultured cells were immunohistochemically stained by monoclonal anti-pan cytokeratin. MTT test was employed to investigate the influence of HSA on the cell attachment on polystyrene surface. The cell growth curve of HGE which were cultured in KSFM with 50 g/L HSA was observed.

RESULTSThe results showed significant decrease in cell numbers within 8 hours after HGE were inoculated, in which the polystyrene surface was preincubated with 50 g/L HSA. But it did not prove to be the case from 10 hours to 24 hours after HGE were inoculated. There were no significant difference within 24 hours in cell numbers between cultured in KSFM with 50 g/L HSA and control. The cell numbers in cell growth curve of HGE in KSFM with and without 50 g/L HSA did not show significant difference.

CONCLUSIONSHSA preincubation on polystyrene were produce inhibitory effect of HGE attachment in early stage.

Cell Adhesion ; drug effects ; Cell Count ; Cell Division ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Gingiva ; cytology ; drug effects ; Humans ; Polystyrenes ; Serum Albumin ; pharmacology

OBJECTIVETo investigate the influence of polysaccharide from Aloe Vera (AP) on the proliferation of the human epithelial cells cultured in vitro.

METHODSThe human epithelial cells undergoing 3 to 4 passages of confluence culture were randomly divided into control and 25, 50, 100, 200 and 400 mg/L AP groups according to different dosage of the polysaccharide (AP) added into the culture medium. In the control group (C), equal volume of DK-SFM medium was added to the culturing cells. The conjugation time of epithelial cells, the changes in the cell morphology and ultrastructure were observed under inverted phase contrast microscope and transmission electron microscope, respectively. The cell proliferation was measured by MTT, cell count analysis and [(3)H]-TdR incorporation. Flow cytometry analysis was employed to detect the cell cycle. The leakage rate of lactate dehydrogenase (LDH) was assayed for the evaluation of the epithelial cell injury.

RESULTSThere was no significant difference in the morphology of the epithelial cells among the groups under inverted phase contrast microscope. But under the transmission electron microscope (TEM), the cells in 100 to 400 mg/L AP groups were seen to have proliferated actively, with euchromatin dominant in the nuclei, while heterochromatin was dominant in the cellular nucleus in control and 25 mg/L AP groups. The confluence time of epithelial cells in 50, 100, 200, 400 mg/L AP groups (154 +/- 12, 141 +/- 20, 130 +/- 19, 124 +/- 13) h preceded noticeably than that in control group (182 +/- 8) h, (P < 0.01). The cell proliferation in 100, 200, 400 mg/L groups reached the peak on the 5th day after AP treatment, while that in control and other groups was delayed by 1 to 2 days. The survival rate of the cells in 25 to 400 mg/L AP groups increased dramatically compared with that in control group, with its [(3)H]-TdR incorporation levels significantly increased in a dose dependent manner. The leakage rate of LDH in 200 and 400 mg/L AP groups was lower than that in control group (P < 0.01). The flow cytometric analysis of the cell cycle distribution revealed that the percentage of cell cycle from phase G0/G1 to G2/M and S in 25 to 400 mg/L AP groups increased significantly in a dose dependent manner compared with that in control group (P < 0.01).

CONCLUSIONAP might be beneficial to the protection of epithelial cells by promoting cell proliferation through inducing the progression of epidermal cells from phase G0/G1 into G2/M and S phases.

Aloe ; Cell Cycle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Humans ; Polysaccharides ; pharmacology