OBJECTIVETo evaluate the protective effect of licoflavone on gastric mucosa in rats with chronic superficial gastritis and explore the possible mechanism.

METHODSSD rat models of chronic superficial gastritis was established by intragastric administration of 0.02% ammonia and long-term irregular diet. The rat models were then randomized into model group, vitacoenzyme group and 3 licoflavone groups of high, medium, and low doses. After 30 days of treatment, the gastric histopathology, mucosal lesions, scanning electron microscopy, mucin function production by the gastric mucosa epithelial cells, serum PGE(2) level and gastric microcirculation were assessed to evaluate the protective effect of licoflavone on gastric mucosa.

RESULTSCompared with normal control rats, the rat models of chronic superficial gastritis showed significantly higher gastric mucosal injury rate, histopathological scores and gastric mucin content. Licoflavone significantly ameliorated gastric pathology and increased serum PGE(2) level, enhanced acidic mucin secretion by the epithelial cells, and improved gastric microcirculation in the rat models.

CONCLUSIONLicoflavone feeding suppresses gastric mucosa injury, protects and restores the injured mucosa in rats with chronic superficial gastritis, and these effects are related with the up-regulation of serum PGE(2) level.

Animals ; Chronic Disease ; Dinoprostone ; blood ; Epithelial Cells ; secretion ; Female ; Flavones ; pharmacology ; Gastric Mucosa ; drug effects ; pathology ; Gastritis ; pathology ; Male ; Microcirculation ; Mucins ; secretion ; Rats ; Rats, Sprague-Dawley

Cell Line ; Cyclooxygenase 2 ; metabolism ; Dinoprostone ; genetics ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Mucin 5AC ; genetics ; secretion ; RNA, Messenger ; genetics ; secretion ; Respiratory System ; cytology ; Tumor Necrosis Factor-alpha ; pharmacology ; Up-Regulation ; drug effects

To explore the role of regulatory peptides in the secretion of bronchial epithelial cells (BECs), we observed the effects of four peptides, i.e.vasoactive intestinal peptide (VIP), epidermal growth factor (EGF), endothelin-1 (ET-1), and calcitonin gene-related peptide (CGRP), on the secretion of ILs from unstimulated or O3-stressed BECs. The results of the experiments showed that VIP exerted an inhibitory effect on the secretion of IL-1 and IL-8 from unstimulated and O3-stressed BECs, VIP also decreased the secretion of IL-5 from O3-stressed BECs; EGF promoted secretion of IL-1 and IL-8 from unstimulated BECs, but decreased the secretion of ILs from O3-stressed BECs; ET-1 and CGRP enhanced the secretion of IL-1, IL-5, and IL-8 from unstimlated BECs, CGRP also increased the secretion of ILs from O3-stressed BECs. The results obtained demonstrate that intrapulmonary regulatory peptides modulate the secretion of ILs from BECs, and may play an important part in transduction of inflammatory signals.
Animals ; Bronchi ; cytology ; Calcitonin Gene-Related Peptide ; pharmacology ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Epidermal Growth Factor ; pharmacology ; Epithelial Cells ; drug effects ; secretion ; Female ; Interleukins ; secretion ; Male ; Rabbits ; Vasoactive Intestinal Peptide ; pharmacology

BACKGROUND: Hypersecretion of mucin due to goblet cell hyperplasia is frequently encountered in many chronic airway diseases, such as chronic bronchitis, bronchiectasis, bronchial asthma and cystic fibrosis. Even in normal individuals, viral infection or bacterial pneumonia frequently provoke huge amounts of bronchial secretions which may cause airway obstruction. The production of mucin was regulated by epidermal growth factor (EGF) in vitro. To know whether this EGF system regulates mucin secretion in vivo and Pseudomonas also stimulates the mucin secretion by the same pathway, we studied these relationships in the cultured rat tracheal epithelial cells. METHODS: Rat tracheal epithelial cells were obtained by pronase dissociation from the male Fisher 344 rats. When cells became confluent, they were divided into 6 groups and stimulated with either EGF for 24 hours or Pseudomonas extracts for 12 hours with or without selective EGF-R tyrosine kinase inhibitor tyrphostin AG1478. RESULTS: We found that both EGF and Pseudomonas extracts phosphorylated the tyrosine residue in the EGF receptor from the rat tracheal epithelial cells and this tyrosine phosphorylation was nearly completely blocked by selective EGF-R tyrosine kinase inhibitor tyrphostin AG1478. The mucin secretion was also stimulated by either EGF or Pseudomonas extracts but more strong secretion of mucin and MUC5AC gene expression in the rat tracheal epithelial cell was done by Pseudomonas extracts. CONCLUSION: These data suggest that Pseudomonas secretes the mucin by way of the EGF receptor and MUC5AC gene expression and the inhibitors of EGF receptor tyrosine phosphorylation would be useful to prevent the huge production of mucin due to Pseudomonas aeruginosa lung infection.
Animal ; Blotting, Western ; Cells, Cultured ; Comparative Study ; Epidermal Growth Factor/*metabolism/pharmacology ; Epithelial Cells/drug effects/*secretion ; Gene Expression ; Male ; Models, Animal ; Mucins/drug effects/*genetics/*secretion ; *Pseudomonas aeruginosa ; RNA, Messenger/analysis ; Rats ; Rats, Inbred F344 ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Trachea/cytology/drug effects/*microbiology/*secretion

OBJECTIVENaringenin has been reported to attenuate Mucin (MUC) 5AC secretion in many pathological models. Many stimuli activate MUC5AC expression through JNK/AP-1 signaling pathways. We hypothesized that naringenin may have inhibitory effects on mucous hypersecretion by modulating MUC5AC production and inhibiting JNK/AP-1 signaling pathways.

METHODThe cell model of mucous hypersecretion was made by human lung adenocarcinoma epithelial (A549) cells stimulated by RSV. A549 cells were subcultured and then randomly divided into 7 groups, which were designated as group C (cell control group), groups R1-3 (cells were infected with RSV at the multiplication of infection (MOI) of 0. 5, 1. 0, 5. 0), groups N1-2 (cells infected with viruses in presence of Nar 30 - 100 mol/L), groups N3-4 (uninfected cells treated with Nar 30 - 100 µmol/L), group D (DMSO), group S (cells infected with viruses in presence of SP600125). After incubating for 24 hrs, the expression of MUC5AC at mRNA and protein level in the groups were determined by real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA). The protein expression changes of JNK, p-JNK and AP-1 were measured by Western blotting.

RESULTThe expressions of MUC5AC protein and mRNA in all RSV infected groups were significantly higher than that in group C in a dose-dependent manner (all P <0. 05). Nar of 30 and 100 µmol/L significantly and dose-dependently decreased RSV-induced secretion of MUC5AC protein in cell supernatant and expression of MUC5AC mRNA (P <0. 05). The relative content of p-JNK, AP-l in R2 groups were 3. 31 ± 0. 34 and 1. 94 ± 0. 05. Theyfrweremtgnificanty increased as compared with group C (both 1. 00 ± 0. 00) (all P <0. 05). The levels of p-JNK in N2 and S groups were 2. 10 ± 0. 20. 27 and 1.±97 ± 0. 16. The levels of AP-1 in N2 and S groups were 1. 40 ± 0. 03, 1. 36 ± 0. 05. Nar and SP600125 led to a largest decrease in levels of p-JNK and AP-1 when compared with group R2 (P <0. 05). The MUC5AC protein in group R2 was (48. 19 ± 0. 47) µg/L. The protein expression of MUC5AC in group R2 was significantly higher than that in group C [(36. 67 ± 1. 50) g/L] with a statistically significant difference (P <0. 05). The protein expression of MUC5AC in groups N2 and S were(43. 17 ± 1. 06) µg/L, (44.±02 ± 0. 99) µg/L, Nar and SP600125 remarkably inhibited RSV-induced secretion of MUC5AC in supernatant of A549 cells (P < 0. 05).

CONCLUSIONSNaringenin might be able to block RSV-induced mucous

Adenocarcinoma ; Blotting, Western ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; Flavanones ; pharmacology ; Humans ; Lung Neoplasms ; Mucin 5AC ; secretion ; Mucus ; secretion ; Random Allocation ; Signal Transduction ; Transcription Factor AP-1 ; drug effects

OBJECTIVETo investigate the impact of human peritoneal mesothelial cells (HPMC) on angiogenesis factor expression and secretion of ovarian carcinoma cell line SKOV3.

METHODSThe conditioned medium with HPMC was tested by ELISA for tumor necrosis factor-alpha (TNF-alpha) and interleukin 10 (IL-1beta). Millicell was used to co-culture HPMC and ovarian carcinoma cell line SKOV3 in the presence or absence of neutralizing antibody against TNF-alpha or IL-1beta. RT-PCR was used to detect vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) gene expression in SKOV3 cells. VEGF and bFGF protein levels in the SKOV3 conditioned medium were assessed by ELISA.

RESULTSConditioned medium with HPMC contained both TNF-alpha and IL-1beta. SKOV3 co-cultured with HPMC expressed higher levels of VEGF and bFGF mRNA and secreted at increased levels of both VEGF and bFGF, in comparison with those in SKOV3 cells cultured alone (P < 0.01). Addition of neutralizing antibody against TNF-alpha or IL-1beta during co-cultures resulted in decrease in mRNA expression and secretion of VEGF and bFGF in SKOV3 cells. When both antibodies were administered during co-culture, additive decrease was observed.

CONCLUSIONHPMC can act in a paracrine fashion to stimulate ovarian tumor cells to produce and secret at increased levels of VEGF and bFGF through TNF-alpha and IL-1beta, and contribute to angiogenesis and peritoneal metastasis of ovarian cancer.

Antibodies ; pharmacology ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Culture Media, Conditioned ; metabolism ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; secretion ; Female ; Fibroblast Growth Factor 2 ; genetics ; secretion ; Gene Expression ; drug effects ; Humans ; Interleukin-1beta ; immunology ; secretion ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Peritoneal Cavity ; cytology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; immunology ; secretion ; Vascular Endothelial Growth Factor A ; genetics ; secretion

No abstract available.
Adenosine Triphosphate/pharmacology ; Animal ; Anions/metabolism ; Biological Transport/physiology ; Biological Transport/drug effects ; Calcium/metabolism* ; Cell Line ; Cell Polarity/physiology* ; Cells, Cultured ; Enzyme Inhibitors/pharmacology ; Epithelial Cells/secretion* ; Epithelial Cells/metabolism ; Epithelial Cells/cytology* ; Fluorescent Dyes ; Fura-2 ; Horses ; Receptors, Purinergic P2/metabolism* ; Thapsigargin/pharmacology

OBJECTIVETo investigate the effect of puerarin on interleukin (IL)-8 mRNA expression and the protein release in the co-culture of human bronchial epithelial (BEAS-2B) cells and human neutrophils.

METHODSBEAS-2B cells and neutrophills were cultured separately and co-cultured with puerarin (50, 100, and 200 μg/mL) for a predetermined time. Cytokines in culture supernatant were evaluated by protein array and IL-8 quantified by enzyme-linked immunosorbent assay (ELISA). IL-8 mRNA expression was evaluated by real-time quantitative polymerase chain reaction (real-time qPCR).

RESULTSThe co-culture of BEAS-2B cells and neutrophils exhibited synergistic effects on IL-8 mRNA expression in BEAS-2B cells, but not in neutrophils after 12 h incubation (P<0.01), as compared with that in BEAS-2B cells or neutrophils alone. IL-8 protein release in the culture supernatant was obviously elevated when BEAS-2B cells were co-cultured with human neutrophils as compared with that in the supernatant of BEAS-2B cells or neutrophils alone after incubated for 2, 6, 12, and 18 h (P<0.01). Treatment with puerarin could significantly down-regulate the expression of IL-8 mRNA in BEAS-2B cells and IL-8 release in the supernatant of the co-culture of BEAS-2B cells and neutrophils (P<0.01).

CONCLUSIONPuerarin could exhibit anti-inflammatory activity by suppressing IL-8 production from the co-culture of human bronchial epithelial cells and neutrophils.

Adult ; Bronchi ; cytology ; Cell Communication ; drug effects ; Cell Line ; Coculture Techniques ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Fluorescence ; Gene Expression Regulation ; drug effects ; Humans ; Interleukin-8 ; genetics ; secretion ; Isoflavones ; pharmacology ; Neutrophils ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction

Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.
Animals ; Benzoates ; pharmacology ; Cystic Fibrosis Transmembrane Conductance Regulator ; antagonists & inhibitors ; drug effects ; metabolism ; Dose-Response Relationship, Drug ; Epithelial Cells ; metabolism ; Exocrine Glands ; secretion ; Flavones ; administration & dosage ; pharmacology ; Humans ; Mice ; Patch-Clamp Techniques ; Rats ; Rats, Inbred F344 ; Thiazolidines ; pharmacology ; Thyroid Gland ; cytology ; Time Factors ; Trachea ; secretion

AIMTo study the mechanisms of anti-diabetic nephropathy of rhein on cultured human mesangial cells (HMCs).

METHODSTo mimic the hyperglycemic (HG) environment of diabetic nephropathy, 30 mmol x L(-1) glucose were added to 10% FBS RPMI 1640. The HMCs were treated with rhein for 8, 24, 48 or 72 h, at these time, the bioactivity, total activity of transforming growth factor-beta1 (TGFbeta1), activity of p38MAPK (p38 mitogen-activated protein kinases, by using immunoprecipitate and Western blot), MMP-2 (matrix metalloproteinase-2), and MMP-9 (matrix metalloproteinase-9, by using gelatinase zymography) and the proliferation of HMCs in high glucose media were measured. Meanwhile the levels of secretion of FN in cultured HMCs were measured.

RESULTSThe results showed that rhein markedly inhibit the proliferation of HMCs, significantly reduce the bioactivity of TGFbeta1 and FN secretion in HMCs, and decrease the increased activity of p38MAPK, but showed no action on the activities of MMP-2 and MMP-9.

CONCLUSIONRhein reduced the secretion of FN and inhibited the proliferation of HMCs may through inhibiting the bioactivities of TGFbeta1 and p38MAPK.

Animals ; Anthraquinones ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; Epithelial Cells ; cytology ; metabolism ; Fibronectins ; secretion ; Glomerular Mesangium ; cytology ; metabolism ; Glucose ; antagonists & inhibitors ; pharmacology ; Humans ; Lung ; cytology ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mink ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; p38 Mitogen-Activated Protein Kinases ; metabolism