Astragalus membranaceus ; chemistry ; Cell Line ; Cytokines ; metabolism ; Humans ; Inflammation ; Macrophages ; drug effects ; metabolism ; Monocytes ; drug effects ; metabolism ; Polysaccharides ; pharmacology

OBJECTIVETo investigate the influence of microcystin-LR (MC-LR) on monocytes and lymphocytes in blood of mice and to find a sensitive index of toxic effects.

METHODSSpecific pathogen free Kunming male mice, aging 1 month-old,were randomly divided into 5 groups by weights, 7 mice for each group. The mice in 5 groups were exposed to MC-LR through intraperitoneal injection at 0, 3.125,6.250, 12.500 and 25.000 µg/kg respectively for 7 days. Then cytokine levels in the serum were measured by radioimmunoassay, DNA-protein crosslinks (DPC) was measured by the SDS/KCl precipitation technique, and the phagocytosis and ROS of leukocytes were detected by flow cytometry.

RESULTSThe levels of interleukin 6 in the 6.250, 12.500 and 25.000 µg·kg(-1)·d(-1) dose groups were (346.837 ± 25.536), (360.847 ± 37.886) and (434.245 ± 35.858)pg/ml respectively, which were significantly higher than those in the control group which the value was (232.775 ± 32.816) pg/ml (t values were -7.258, -6.760 and -10.966 respectively, P values were all < 0.05).While the level of tumor necrosis factor-alpha was(10.782 ± 0.966) fmol/ml in 25 µg·kg(-1)·d(-1) dose group was statistically lower than it in the control group which the value was (16.878 ± 3.378) fmol/ml (t value was 4.591, P < 0.05). The DPC levels of lymphocytes in 6.250, 12.500 µg·kg(-1)·d(-1) dose group were (242.576 ± 7.545),(241.472 ± 2.793) ng/ml,higher than it in the control group while the value was (228.657 ± 4.130) ng/ml (t value was -4.282, -6.801, P values were all <0.05). The fluorescence intensity of DCF in lymphocytes in the 4 treated groups were separately 3299.37 ± 120.54, 3281.38 ± 58.34, 3308.06 ± 136.12 and 3346.92 ± 108.69, all significantly lower than 3770.81 ± 131.39 in the control group (t values were 6.995, 9.007, 6.472 and 6.577 respectively, and P values were all <0.05). The fluorescence intensity of DCF in monocytes in the 4 treated groups (3271.51 ± 140.79, 3270.05 ± 117.92, 3326.90 ± 114.39 and 3292.49 ± 145.97 respectively) were also significantly lower than the value in the control group was 3841.72 ± 130.92 (t values were 7.847, 8.584, 7.835 and 7.411 respectively, P values were all <0.05). There was no significant difference in other index among the four experiment groups and the control group.

CONCLUSIONThe MC-LR administered via intraperitoneal injection to mice induced the alterations of some cytokines of monocytes and lymphocytes in blood. By comparison, the ROS of leukocyte was the most sensitive index.

Animals ; Cytokines ; metabolism ; Lymphocytes ; drug effects ; Male ; Mice ; Mice, Inbred Strains ; Microcystins ; pharmacology ; Monocytes ; drug effects ; Reactive Oxygen Species ; metabolism

During Toxoplasma gondii infection, macrophages, dendritic cells, and neutrophils are important sources of pro-inflammatory cytokines from the host. To counteract the pro-inflammatory activities, T. gondii is known to have several mechanisms inducing down-regulation of the host immunity. In the present study, we analyzed the production of proand anti-inflammatory cytokines from a human myelomonocytic cell line, THP-1 cells, in response to treatment with T. gondii lysate or lipopolysaccharide (LPS). Treatment of THP-1 cells with LPS induced production of IL-12, TNF-alpha, IL-8, and IL-10. Co-treatment of THP-1 cells with T. gondii lysate inhibited the LPS-induced IL-12, IL-8 and TNF-alpha expression, but increased the level of IL-10 synergistically. IL-12 and IL-10 production was down-regulated by anti-human toll-like receptor (TLR)-2 and TLR4 antibodies. T. gondii lysate triggered nuclear factor (NF)-kappaB-dependent IL-8 expression in HEK293 cells transfected with TLR2. It is suggested that immunosuppression induced by T. gondii lysate treatment might occur via TLR2-mediated NF-kappaB activation.
Animals ; Cell Line ; Cytokines/*biosynthesis ; Humans ; Inflammation/metabolism ; Lipopolysaccharides/*pharmacology ; Macrophages/*drug effects/*metabolism ; Toxoplasma

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

OBJECTIVETo discuss the effect of integrated traditional Chinese and Western medicine on Th1/Th2 cytokines level in children with asthma.

METHODChildren with asthma were randomly divided into control group and treatment group. The control group was given Montelukast Sodium tablets for 4 mg (2-5 years old) or 5 mg (6-14 years old), once every night before sleeping. At the same time, the treatment group was given Pingchuan water decoction additionally for one tie per day, in four to six-divided doses for eight weeks. On the other hand, groups of health children were selected as blank control. Before and after treatment, the level of IL-4, IL-13 and IFN-gamma were detected by elisa from 3 mL of venous blood.

RESULTBefore treatment, IL-4 and IL-13 in two groups were obviously higher than healthy group, but IFN-gamma was lower (P < 0.01, P < 0.05). After eight weeks, IL-4 and IL-13 in treatment group went down, but IFN-gamma increased. And there were significant different compared with the control group (P < 0.05).

CONCLUSIONTreating children asthma by integrated traditional Chinese and western medicine can obviously reduce IL4 and IL-13, increase IFN-gamma, and therefore lighten the airway inflammation, prevent asthma attacks effectively.

Adolescent ; Asthma ; drug therapy ; immunology ; metabolism ; Child ; Cytokines ; metabolism ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Th1 Cells ; drug effects ; metabolism ; Th2 Cells ; drug effects ; metabolism ; Western World

OBJECTIVEDendritic cells an hyperinsulinemia are both implicated in the pathogenesis of atherosclerosis. The aim of this study is to explore the effect of high concentration of insulin on the maturation of monocyte-derived dendritic cells (MoDCs) and related signal transduction pathways.

METHODSHuman monocytes were purified (over 98%) using Anti-CD14 micro-beads and cultured for 5 days with DC Cellgro medium containing rhGM-CSF (100 microg/L) and rhIL-4 (20 microg/L). Immature DC were then incubated with insulin of various concentrations (0, 1, 10, 100 nmol/L) for 24 hours in the presence or absence of LY294002 (PI3K inhibitor) or PD98059 (MAPK inhibitor). Immunophenotypic expression of CD86 and CD83 were detected using flow cytometry. Endocytosis function of the MoDCs was evaluated using FITC-Dextran and MoDCs secretion IL-12, IFN-gamma and TNF-alpha were measured by ELISA.

RESULTSInsulin induced significantly higher CD83 and CD86 expressions on MoDCs in a dose-dependent manner. The endocytosis function of MoDCs were significantly inhibited and cytokine secretions of IL-12, IFN-gamma and TNF-alpha significantly increased by 10 nmol/L and 100 nmol/L insulin. These effects could be blocked by the LY294002 and PD98059.

CONCLUSIONHyperinsulinemia contributed to atherosclerosis via stimulating immune maturation of MoDCs via both PI3K and MAPK pathways.

Cell Differentiation ; drug effects ; immunology ; Cells, Cultured ; Cytokines ; metabolism ; Dendritic Cells ; drug effects ; immunology ; metabolism ; Humans ; Insulin ; administration & dosage ; pharmacology ; Monocytes ; cytology ; Phagocytosis ; drug effects ; Signal Transduction

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

This experimental controlled study was performed to evaluate the composition of autologous processed plasma (APP), and the effects of APP intra-articular injection into healthy equine metacarpophalangeal joints. The effects on joints were analysed with a short-phase protocol and a prolonged-phase protocol using saline-injected joints as controls. For the short protocol, horses received one intra-articular APP injection. Synovial fluid samples were collected prior to the injection and 3, 6, 24, 48, and 16 h after treatment. For the prolonged protocol, the joints received three weekly injections of APP, and samples were collected at 0, 7, 14, 21, and 28 days before APP administration. IL1-ra level was found to be increased in APP compared to plasma. Upon intra-articular administration of APP, transient (up to 24 h) increases in white blood cell (WBC) counts along with elevated protein and prostaglandin E2 (PGE2) concentrations were observed in the treated joints. Over the 28-day observation period, APP did not elicit changes relative to baseline levels, but WBC counts, PGE2 and chondroitin sulphate concentrations were lower than those found in the control. In conclusion, APP intra-articular injection induced a mild and transitory inflammatory response but no inflammation reaction was observed over a longer period of treatment and observation.
Animals ; Cytokines/*metabolism ; *Horses ; Injections, Intra-Articular ; Metacarpophalangeal Joint/*drug effects ; Plasma/*chemistry ; Time Factors

Bone Resorption ; China ; Cytokines ; physiology ; Diphosphonates ; pharmacology ; Estrogens ; pharmacology ; Humans ; Jaw ; drug effects ; metabolism

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.
Animals ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Glucose ; metabolism ; Indole Alkaloids ; pharmacology ; Inflammation ; metabolism ; Insulin Resistance ; Male ; Muscle, Skeletal ; cytology ; drug effects ; metabolism ; Quinazolines ; pharmacology ; Rats