Cell Line ; Glucose ; metabolism ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect of 1,25 (OH)(2)D(3) on cell proliferation and the expression of tumor necrosis factor-α (TNF-α ) in human gastric carcinoma SGC-7901 cells.

METHODSSGC-7901 cells were treated with 1×10(-6), 1×10(-7), 1×10(-8), and 1×10(-9) mol/L 1,25 (OH)(2)D(3) for 24, 48, 72 and 96 h. The cell proliferation was measured by MTT assay, and the cell cycle changes were analyzed using flow cytometry. RT-PCR and Western blotting were used to determine the expression of TNF-α mRNA and protein, respectively.

RESULTS1,25 (OH)(2)D(3) significantly inhibited SGC-7901 cell proliferation (P<0.05) in a time- and dose-dependent fashion. Treatment with 1,25 (OH)(2)D(3) for 72 h caused significant cell cycle arrest at G(0)/G(1) phase (F=9.81, P<0.05) and dose-dependently inhibited the expression of TNF-&alpha; at both mRNA and protein levels in SGC-7901 cells (P<0.05).

CONCLUSIONThe inhibitory effect of 1,25 (OH)(2)D(3) on SGC-7901 cell proliferation is probably associated with the down- regulation of TNF-&alpha; expression.

Calcitriol ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; metabolism

Anti-Inflammatory Agents ; pharmacology ; Brain ; drug effects ; metabolism ; Humans ; NF-kappa B ; metabolism ; Neurons ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; alpha-MSH ; pharmacology

OBJECTIVETo study the possible induction effect of exposure to 50 Hz magnetic field (MF) on clustering of cell membrane surface receptors for epidermal growth factor (EGF) and tumor necrosis factor (TNF), the starting site of signals of biological effects, and its possible intervention effect.

METHODSLung fibroblasts of Chinese hamster (CHL) were exposed to EGF, TNF, 0.4 mT 50 Hz MF, 0.4 mT noise MF, and 0.4 mT 50 Hz MF combined with 0.4 mT noise MF. Respectively, for different durations, following the treatment, EGF and TNF receptors on the cell membrane were marked by corresponding antibodies with immunohistochemical method, then observed under a confocal microscope.

RESULTSClustering of cell membrane receptors could be induced 5 min after treatment with EGF and TNF, as well as with 50 Hz MF at 0.4 mT, which reached the peak in 15 min. While noise MF with the same intensity did not induce clustering of cell membrane receptors. Superposition of noise MF with the same intensity could inhibit clustering of cell membrane receptors induced by 50 Hz MF.

CONCLUSIONClustering of EGF and TNF receptors on the cell membrane could be induced by 50 Hz MF, suggesting that membrane receptors would be one of the sites where MF signals coupled, and noise MF with the same intensity could inhibit these effects.

Animals ; Cell Line ; Cricetinae ; Electromagnetic Fields ; adverse effects ; Epidermal Growth Factor ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; radiation effects ; Noise ; adverse effects ; Receptor, Epidermal Growth Factor ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

BACKGROUNDInterleukin (IL)-1beta may effectively decrease introcular pressure (IOP) when administered by subconjunctival injection in normal rabbit. However, IL-1beta is a large molecular agent and an inflammation factor. The aim of this study was to evaluate the penetrability of IL-1beta, and the concentrations of both tumor necrosis factor (TNF)-alpha and IL-6 in the aqueous humor of normal rabbits treated with IL-1beta.

METHODSA total of 170 rabbits were used in the study and were assigned to several different treatment groups as follows: 125 of the rabbits were assigned to two groups. In one group, 33 rabbits were injected subconjunctivally with IL-1beta and 39 were injected with saline alone. In the other group, 27 rabbits were given eye drops containing IL-1beta (400 ng/ml) and 26 were given saline alone. Aqueous humor (AH) was drawn and the concentration of IL-1beta within the fluid measured. The IOP was measured in another six rabbits after administration of eye drops containing IL-1beta (400 ng/ml). A further 20 rabbits were assigned to 3 groups as follows: eight untreated normal controls; six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of TNF-alpha in the fluid was measured. Another 19 rabbits were assigned to 3 groups as follows: seven untreated normal controls; and six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of IL-6 in the fluid measured. Measurement of cytokine concentration was by radio-immunoassay in all cases.

RESULTSThe IL-1beta concentration in the AH was higher in those animals in which it had been administered subconjunctivally (P < 0.01). The IL-1beta concentration in the AH of the animals given eye drops was almost the same as that in the controls (P > 0.05). The administration of IL-1beta in the form of eye drops had little effect upon IOP reduction. Lower TNF-alpha concentrations were seen in the AH after the subconjunctival administration of IL-1beta, but the concentration of IL-6 was the same as in the normal controls.

CONCLUSIONSIL-1beta shows good corneal penetrability after subconjunctival injection into normal rabbit eyes. The IOP reduction induced by IL-1beta is unlikely be associated with an inflammatory response.

Animals ; Aqueous Humor ; drug effects ; metabolism ; Interleukin-1beta ; pharmacology ; Interleukin-6 ; metabolism ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore the effects of TNF alpha on the expression of sterol regulatory element binding proteins cleavage activating protein (SCAP) and triglyceride contents in cells of a model of cultured steatotic hepatocytes.

METHODSA steatotic hepatocytes model was established by treating L-02 cell strain with oleic acid. The cells were treated with TNF alpha and/or TNF alpha antibody. The cells were divided into six groups: a control group (C), a model group (F), a control group with TNF alpha (C1), a control group with TNFalpha antibody (C2), a model group with TNFalpha(F1) and a model group with TNFalpha antibody (F2). The expression of SREBP-1c mRNA was measured with RT-PCR; the protein expression of SCAP was measured by Western blot; lipid droplets in the hepatocytes were observed with oil red O staining; the contents of triglyceride in hepatocytes were measured with an analytical kit.

RESULTSThe mRNA expression of SCAP in the groups treated with TNF alpha were upregulated compared with those of the control group (C1 vs C increased 67%, F1 vs F increased 55%, F = 212.98), the protein expression of SCAP in the groups treated with TNF alpha was upregulated compared with those of the control group (C1 vs C increased 45%, F1 vs F increased 95%, F = 104.3), and triglyceride contents in hepatocytes of these groups were increased compared with those of the control group [C (2.02+/-0.67) mg/10(7) cells, F(7.79+/-1.35) mg/10(7) cells, F1(13.36+/-1.99) mg/10(7) cells, F = 82.94].

CONCLUSIONTNF alpha upregulates the expression of SCAP and promotes the synthesis of triglyceride; it probably participates in the process of developing steatosis of hepatocytes.

Cell Line ; Hepatocytes ; drug effects ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Membrane Proteins ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the TNF-alpha induced apoptosis of U937 cells, the expression, degradation and subcellular localization of IkappaB-alpha, and its degradation mechanism.

METHODChanges and subcellular loca-lization of IkappaB-alpha were observed by fluorescence microscopy, expression and degradation of IkappaB-alpha protein with N-tosyl-L-phenylalanylchloromethyl ketone (TPCK protease inhibitor) blocking test and apoptosis of U937 cell by flow cytometry.

RESULTS(1) immunolfluorescence assay showed that IkappaB-alpha localized in cytoplasm only. (2) The level of IkappaB-alpha protein was downregulated after TNF-alpha stimulation, flow cytometry also confirmed the downregulation. (3) The downregulation of IkappaB-alpha protein levels in TNF-alpha induced apoptosis was partially inhibited by TPCK. (4) The apoptosis rate of U937 cells induced by TNF-alpha was (60.73 +/- 1.61)%.

CONCLUSION(1) Degradation of IkappaB-alpha protein during TNF-alpha induced apoptosis of U937 cells suggested the activation of NF-kappaB. (2) TPCK sensitive protease plays an important role in the degradation of IkappaB-alpha protein. (3) TPCK sensitive protease also involved in the apoptosis of U937 cells induced by TNF-alpha.

Apoptosis ; drug effects ; Down-Regulation ; Humans ; NF-kappa B ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; U937 Cells

OBJECTIVETo clarify the protective roles of compatibility of geniposide and ginsenoside (Rg1) in regulating ischemia injured microglia homeostasis by comparing the difference in regulatory roles of geniposide, Rg1, or ginsenoside + Rg1 in balancing secretion of oxygen glucose deprivation induced microglia inflammatory cytokines.

METHODSThe mimic ischemia injured microglia model was induced by oxygen-glucose deprivation (OGD). Then geniposide, Rg1, or ginsenoside + Rg1 (Tongluo Jiunao Injection, TJI) was respectively added. The NO content was determined by Griess Reagent. The cyto activity was detected using cell count kit. Contents of TNF-alpha and TGF-beta and their expression levels were detected by ELISA and Western blot.

RESULTSGeniposide + Rg1 could significantly inhibit the release of NO, elevate the TGF-beta level, and decrease the content of TNF-alpha without influencing the cell survival. The two active ingredients played different therapeutic roles. The compatible use was obviously superior to use any one of the two active ingredients alone.

CONCLUSIONSGeniposide, Rg1, or Ginsenoside + Rg1 had regulating roles in balancing ischemia injured microglia homeostasis. Its mechanisms might be related to up-regulating the TGF-beta expression and down-regulating TNF-alpha expression.

Animals ; Ginsenosides ; pharmacology ; Hypoxia ; metabolism ; Iridoids ; pharmacology ; Mice ; Microglia ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect of formaldehyde exposure on the expression of inflammatory cytokines in human bronchial epithelial cells (16HBE cells).

METHODS16HBE cells were treated with formaldehyde with a concentration of 0, 0.04, 0.08, 0.16, 0.32, or 0.64 mmol/L for 24 hours, and MTT assay was applied to measure proliferative activity and calculate median lethal dose; 16HBE cells were exposed to formaldehyde with a concentration of 0, 0.04, 0.16, 0.64, or 1.20 mmol/L for 4 hours, MTT assay was applied to measure proliferative activity, and enzyme-linked immunosorbent assay was applied to measure the levels of Th1, Th2, and Th17 cytokines and tumor necrosis factor &alpha;(TNF-&alpha;) in cell supernatant.

RESULTSCompared with the control group, the 0.32-and 0.64-mmol/L exposure groups had significant decreases in cell viability (P<0.05); all exposure groups had reductions in interleukin(IL)-2 and IL-12, but no significant changes in interferon-γ and IL-10. In the 1.20-mmol/L exposure group, there was an increase in IL-4, with the increasing exposure dose, IL-5 and IL-6 tended to increase first and then decrease, and there was no significant change in IL-13; with the increasing exposure dose, IL-8 tended to increase first and then decrease, and there was no significant change in IL-17. In all the exposure groups, TNF-&alpha; increased and tended to increase significantly with the increasing exposure dose(P<0.05).

CONCLUSIONFormaldehyde exposure can cause imbalance between Th1 and Th2 cytokines secreted by 16HBE cells, as well as increased expression of IL-8 and TNF-&alpha;.

Cells, Cultured ; Cytokines ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; Formaldehyde ; adverse effects ; Humans ; Interferon-gamma ; metabolism ; Interleukins ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Animals ; Immunoglobulin A, Secretory ; metabolism ; Intestines ; drug effects ; metabolism ; Male ; Massive Hepatic Necrosis ; metabolism ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; pharmacology