BACKGROUNDThe pathogenesis of acute pancreatitis is complex and largely unclear. The aim of this study was to explore the relationship between modes of cell death in pancreatic acinar cells, the release of cell contents and the inflammatory response of macrophages.

METHODSOur experiment included four groups: group A (the control group), group B (AR42J cells overstimulated by caerulein), group C (AR42J cells treated with lipopolysaccharide and caerulein), and group D (AR42J cells treated with octreotide and caerulein). Apoptosis and oncosis, and the release of amylase and lactate dehydrogenase (LDH) from AR42J cells were detected. Rat macrophages were stimulated by 1 ml supernatant of culture medium of AR42J cells. Finally, NF-kappaB activation and TNF-alpha and IL-1beta secretion by macrophages were detected.

RESULTSOncotic cells in group C increased while apoptotic cells decreased (P < 0.05); cells in group D had the inverse reaction. The release of amylase and LDH changed directly with the occurrence of oncosis. The transcription factor NF-kappaB was activated and secretion of TNF-alpha and IL-1beta were significantly higher in group C than in group B (P < 0.05); in group D, these actions were significantly lower than in group B (P < 0.05). This trend was in line with changes in amylase and LDH production.

CONCLUSIONThere is a close relationship between modes of pancreatic acinar cell death, the release of cell contents and the inflammatory reaction of macrophages.

Amylases ; secretion ; Animals ; Apoptosis ; Interleukin-1beta ; secretion ; L-Lactate Dehydrogenase ; secretion ; Macrophage Activation ; Male ; NF-kappa B ; metabolism ; Pancreas ; pathology ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo detect the expression of cytokines by acute promyelocytic leukemia (APL) cells before and after exposure to arsenic trioxide.

METHODSDiagnoses were performed according to the FAB cytological classification criteria and cytogenetic criteria. Bone marrow or blood samples from APL patients were collected in heparinized tubes, then primary APL cells were separated by traditional Ficoll-Hypaque density centrifugation and purified after adherence to plastic surfaces. IL-1(beta), IL-6, IL-8, TNF alpha and G-CSF levels in the leukemia cell culture supernatants were detected by ELISA. At the same time, nitro blue tetrazolium (NBT) reduction test was used to detect the differentiation of APL cells.

RESULTSAfter 96 hours exposure to arsenic trioxide, 10 - 6 mol/L in vitro or 10 mg/d in vivo, APL cells showed a significant increase of IL-1(beta) (P < 0.05) and G-CSF (P < 0.05) production, and a significant decrease of IL-6 (P < 0.05) and IL-8 (P < 0.05). However, there was no obvious variation of TNF alpha when compared with APL cells without exposure to arsenic trioxide. On the other hand, the proliferation ratio of APL cells in vitro was statistically correlated to the IL-1(beta) secretion ratio or G-CSF secretion ratio. The cell number ratio in patients with detectable IL-1(beta) or G-CSF was higher than that without detectable IL-1(beta) or G-CSF.

CONCLUSIONIL-1(beta) and G-CSF secretion may play an important role in the proliferation of APL cells after exposure to arsenic trioxide.

Arsenicals ; pharmacology ; Cells, Cultured ; Cytokines ; secretion ; Granulocyte Colony-Stimulating Factor ; secretion ; Humans ; Interleukin-1 ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Leukemia, Promyelocytic, Acute ; metabolism ; Oxides ; pharmacology ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo detect the changing profiles of three factors in nasal fluid before and after endoscopic sinus surgery (ESS), including platelet derived growth factor (PDGF), tumour necrosis factor-alpha (TNF-alpha) and hyaluronic acid (HA), to investigate the predictive factor of healing quality after sinus surgery.

METHODSNasal secretions were obtained from 22 patients with nasal polyp before ESS and at 1, 4, 8, 12 weeks after ESS compared to 22 controls. Nasal fluid was analyzed by enzyme-linked immunosorbent assay (ELISA) for PDGF, TNF-alpha and HA. The healing quality was evaluated at 1, 4, 8, 12 weeks postoperatively for every one. Finally, we performed a multiple logistic regression model to assess whether the healing quality was associated with age, gender, body mass, preoperative stage, and concentrations of PDGF, TNF-alpha, HA before ESS.

RESULTS1. Each factor before surgery showed different level when compared to controls, PDGF was constant, TNF-alpha was high (P = 0.034), and HA was low (P = 0.003). Comparing patients at 1 week after surgery with control subjects, a significant increase in the concentrations of PDGF, TNF-alpha, HA in nasal secretions could be demonstrated (compared to controls, P = 0.000, 0.020, 0.038, respectively). After that, decreasing amounts of these factors were found, reaching normal conditions at 12 weeks for PDGF (compared to controls, P = 0.087), 8 weeks for TNF-alpha (compared to controls, P =0.104), 4 weeks for HA (compared to controls, P = 0.304). There was another peak for TNF-alpha at 12 weeks (compared to controls, P = 0.002). 2. The percentage of good healing at 1, 4, 8, 12 week postoperatively was 0, 4.5%, 36.4%, 81.8%, respectively. The healing quality after ESS was significantly and independently correlated to the age of patient and preoperative PDGF concentrations in nasal secretions. The younger, the better healing. The lower PDGF concentration, the better healing.

CONCLUSIONSDuring the wound healing of nasal mucosa, the levels of PDGF, TNF-alpha, HA were different at each postoperative stage. Age and PDGF concentrations preoperatively were suitable factors to predict the healing quality after ESS.

Adolescent ; Adult ; Aged ; Body Fluids ; secretion ; Case-Control Studies ; Endoscopy ; Female ; Humans ; Hyaluronic Acid ; secretion ; Male ; Middle Aged ; Nasal Mucosa ; metabolism ; secretion ; Nasal Polyps ; metabolism ; surgery ; Platelet-Derived Growth Factor ; secretion ; Tumor Necrosis Factor-alpha ; secretion ; Young Adult

OBJECTIVETo study the effects of Aloe coarse polysaccharide on the levels of growth factors (EGF, TGF-alpha, TGF-beta1) and interleukins (IL-1beta, IL-6, IL-8) and tumor necrosis factor (TNF) in cultured keratinocytes.

METHODThe cultured keratinocytes were treated with Aloe coarse polysaccharide at concentrations of 75, 150, 300, 600, 1 200 mg x L(-1) land the equal volume of media as control group. The levels of EGF, TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were assayed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA).

RESULTCompared with the control group, the levels of EGF, TGF-alpha, IL-1beta, IL-6 and IL-8 were significantly increased by treatment with Aloe coarse polysaccharide (P < 0.05, P < 0.01) and in a dose dependent manner, and the levels of TGF-beta1 and TNF were also increased but no statistical significance.

CONCLUSIONAloe coarse polysaccharide may promote keratinocytes to secrete EGF, TGF-alpha, IL-1beta, IL-6 and IL-8.

Aloe ; chemistry ; Cells, Cultured ; Cytokines ; secretion ; Dose-Response Relationship, Drug ; Epidermal Growth Factor ; secretion ; Humans ; Interleukin-1beta ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Keratinocytes ; secretion ; Plants, Medicinal ; chemistry ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacology ; Transforming Growth Factor alpha ; secretion ; Transforming Growth Factor beta1 ; secretion ; Tumor Necrosis Factors ; secretion

OBJECTIVETo investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on secretion of inflammatory cytokines in macrophages stimulated by lipopolysacharide (LPS).

METHODSRat BMSCs and macrophages were isolated, cultured, and identified. The BMSCs and macrophages, cultured alone or in co-culture, were treated with LPS or PBS or without treatment and tested for interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) concentrations in the supernatants at 1, 3, 5, 7, 12, and 24 h after the treatment using enzyme-linked immunosorbent assay.

RESULTSExposure to LPS caused significantly increased IL-10 and TNF-α concentrations in the supernatant of cultured macrophages but not in BMSC culture. Macrophages co-cultured with BMSCs showed significantly lowered IL-10 and TNF-α secretions in response to LPS exposure as compared with the macrophages cultured alone.

CONCLUSIONBMSCs can reduce LPS-induced secretion of inflammatory cytokines by the macrophages to ameliorate inflammatory reactions.

Animals ; Cells, Cultured ; Coculture Techniques ; Enzyme-Linked Immunosorbent Assay ; Inflammation ; Interleukin-10 ; secretion ; Lipopolysaccharides ; Macrophages ; secretion ; Mesenchymal Stromal Cells ; cytology ; Rats ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo investigate the effect of necrotic cells on the secretion of inflammatory cytokines.

METHODSRAW264.7 macrophages and necrotic mouse thymocytes induced by heating were incubated for 18 h at a ratio of 5:1 in the absence or presence of lipopolysaccharide (LPS, 100 ng/ml). The supernatant of the cell culture was collected and the expression and secretion of the pro-inflammatory cytokines were measured using Bio-Plex suspension system.

RESULTSThe secretions of tumor necrosis factor-alpha (TNF-alpha) and interlukine-6 (IL-6) by macrophages co-cultured with the necrotic cells were significantly enhanced as compared with the control cells. The necrotic cells also significantly augmented the secretion of the pro-inflammatory cytokines induced by LPS.

CONCLUSIONNecrotic cells not only induces pro-inflammatory cytokine expression by themselves but also work synergistically with LPS to enhance the cytokine production, suggesting the important roles of necrotic cells to initiate and maintain the inflammatory responses.

Animals ; Cell Line ; Hot Temperature ; Inflammation ; etiology ; metabolism ; pathology ; Interleukin-6 ; secretion ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Male ; Mice ; Necrosis ; complications ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo investigate the effects of apoptotic lymphocytes on the secretion of cytokines by hepatic sinusoidal endothelial cells (HSEC).

METHODSHuman HSEC cells were co-cultured for 16 h with allogenetic apoptotic lymphocytes induced by UVB irradiation. The supernatants were collected and the levels of interleukin-2, interferon-gamma, and tumor necrosis factor-alpha were detected by Luminex technique.

RESULTSAll the cytokines were down-regulated by about 50% in HSECs after co-culture with the apoptotic lymphocytes as compared with those in the control group (P<0.05).

CONCLUSIONSCo-culture with apoptotic lymphocytes can down-regulate the secretion of pro-inflammatory cytokines in HSECs, which may contribute to tolerogenic microenvironment in the liver.

Apoptosis ; physiology ; Cells, Cultured ; Coculture Techniques ; Cytokines ; secretion ; Down-Regulation ; Endothelial Cells ; cytology ; metabolism ; Humans ; Immune Tolerance ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Liver ; cytology ; Lymphocytes ; cytology ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo study the effect of a new obesity-related gene NYGGF4 on the insulin sensitivity and secretory function of adipocytes.

METHODS3T3-L1 preadipocytes transfected with either an empty expression vector (pcDNA3.1; control group) or an NYGGF4 expression vector (NYGGF4-pcDNA3.1) were cultured in vitro and differentiated into the matured adipocytes with the standard insulin plus dexamethasone plus 3-isobutyl-methylxanthine (MDI) induction cocktail. 2-deoxy-D-[3H] glucose uptake was determined by liquid scintillation counting. Western blot was performed to detect the protein content and translocation of glucose transporter 4 (GLUT4). The supernatant concentrations of TNF-alpha, IL-6, adiponectin and resistin were measured using ELISA.

RESULTSNYGGF4 over-expression in 3T3-L1 adipocytes reduced insulin-stimulated glucose uptake. NYGGF4 over-expression impaired insulin-stimulated GLUT4 translocation without affecting the total protein content of GLUT4. The concentrations of TNF-alpha, IL-6, adiponectin and resistin in the culture medium of 3T3-L1 transfected with NYGGF4 were not significantly different from those in the control group.

CONCLUSIONSNYGGF4 over-expression impairs the insulin sensitivity of 3T3-L1 adipocytes through decreasing GLUT4 translocation and had no effects on the secretory function of adipocytes.

3T3-L1 Cells ; Adipocytes ; drug effects ; secretion ; Adiponectin ; secretion ; Animals ; Carrier Proteins ; genetics ; physiology ; Glucose ; metabolism ; Glucose Transporter Type 4 ; analysis ; metabolism ; Insulin ; pharmacology ; Interleukin-6 ; secretion ; Mice ; Resistin ; analysis ; Transfection ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo determine the effect of Porphyromonas gingivalis lipopolysaccharide (LPS) on induced secretion of inflammatory cytokines by different cell lines.

METHODSLPS of P. gingivalis strain ATCC33277 (Pg-LPS) was extracted with phenol-water method, and identified by Limulus test and infrared spectrum analysis. KB, HGF-1 and THP-1 cells were treated with Pg-LPS of different concentrations and time duration, a commercial LPS of E.coli strain O111:B4 (E-LPS) was used as the control. TNF-alpha, IL-1beta, IL-6 and IL-8 levels in culture supernatants were measured by quantitative ELISA.

RESULTThe minimal dosages of both Pg-LPS and E-LPS to solidify Limulus agents were 15 ng/ml and their infrared spectrums were similar. With the treatment of Pg-LPS or E-LPS, the TNF-alpha and IL-1beta levels secreted by HGF-1 cells were remarkably increased with a single perk (P<0.01) while a continuous enhancement of secretion by THP-1 cells was observed (P<0.01). Either Pg-LPS or E-LPS stimulated HGF-1 or THP-1 cells to continuously increase the secretion of IL-6 (P<0.01). Both Pg-LPS and E-LPS induced IL-8 secretion by THP-1 cells (P<0.01), but only Pg-LPS showed the similar effect on HGF-1 cells (P<0.01). Neither Pg-LPS nor E-LPS induced KB cells to secrete inflammatory cytokines.

CONCLUSIONPg-LPS can promote target cells to increase their secretion of inflammatory cytokines. KB cells can not be used as the target cell to determine inflammation-causing effect of LPS.

Cell Line ; Epithelial Cells ; cytology ; metabolism ; Fibroblasts ; cytology ; metabolism ; Humans ; Interleukin-1beta ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Lipopolysaccharides ; pharmacology ; Porphyromonas gingivalis ; chemistry ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo investigate the effect of FK506 on cytokine secretions in whole blood from healthy individuals.

METHODSBlood samples collected from healthy volunteers were co-cultured with different concentrations of FK506 and stimulated with PMA and IONO. The concentrations of 8 cytokines including IL-2, IL-6, IL-12, IL-17, IFN-gamma, TNF-alpha, GM-CSF and G-CSF were detected by Bio-Plex suspension system.

RESULTSCompared with the control group, high-concentration FK506 (20 ng/ml) significantly inhibited the secretions of IL-2, IL-6, IL-12, IL-17, IFN-gamma, TNF-alpha, GM-CSF and G-CSF. At a moderate concentration (5 ng/ml), FK506 inhibited the secretion of GM-CSF significantly.

CONCLUSIONFK506 effectively inhibits the secretion of proinflammatory cytokines including IL-6, IFN-gamma and TNF-alpha and also the secretion of IL-2, IL-12, IL-17, GM-CSF and G-CSF. FK506 might play the role of immunosuppression by inhibiting the production of these cytokines by the immune cells. Monitoring the levels of these cytokines might be a potential method for evaluating the adequacy of FK506 doses administered.

Adult ; Cytokines ; blood ; secretion ; Down-Regulation ; drug effects ; Female ; Humans ; Immunosuppressive Agents ; pharmacology ; Interferon-gamma ; blood ; secretion ; Interleukin-6 ; blood ; secretion ; Male ; Tacrolimus ; pharmacology ; Tumor Necrosis Factor-alpha ; blood ; secretion ; Young Adult