PURPOSE: We wanted to evaluate the changes of the expression of macrophage migration inhibitory factor(MIF) according to time, so we determined the semiquantitative score from immunostaining in a bladder inflammatory rat model. MATERIALS AND METHODS: A total of 25 female Sprague-Dawley rats were divided into 5 groups according to the time course. Group 1 was the control group that was treated with an intravesical instillation of saline. Groups 2-5 were evaluated at 4 hours, 24 hours, 48 hours and 7 days after the instillation of lipopolysaccharide(LPS), respectively. H&E staining and immunohistochemical staining for MIF were performed after removing the bladder. A semiquantitative score was used to evaluate the cystitis(bladder inflammation score; BIS). The expression of MIF in the bladder was graded from 0 to 3+(MIF score; MIFS). RESULTS: The staining of MIF was the most intense in the basal layer of the urothelium in the control group(BIS 0, MIFS 3). The degree of bladder inflammation was highest in group 3, and MIF was not expressed even in the urotherlium without inflammation(BIS 2.2, MIFS 0.6). The bladder inflammation was decreased after 48 hours, and the expression of MIF was increased after 48 hous(BIS 1.7, MIFS 1.6). The severity of bladder inflammation and the expression of MIS were significantly changed with the time course(p<0.001). CONCLUSIONS: These results suggest that pre-formed MIF is stored in the cytoplasm of the basal cells in the urothelium, and it is released into the lumen of the bladder after a noxious stimulus like LPS.
Administration, Intravesical ; Animals ; Cystitis* ; Cytokines ; Cytoplasm ; Female ; Humans ; Inflammation ; Lipopolysaccharides ; Macrophages* ; Models, Animal ; Rats* ; Rats, Sprague-Dawley ; Urinary Bladder ; Urothelium

The present study was planned to investigate the pharmacokinetics of ceftriaxone in experimentally induced febrile buffalo calves (n = 5). The fever was induced by intravenous injection of E.coli lipopolysaccaride (1 microgram/kg). To study the pharmacokinetics, ceftriaxone was administered at the dose rate of 10 mg/kg body wt. in all animals. At 1 min, the peak concentration of ceftriaxone was 79.4 +/- 2.37 microgram/ml and the drug was detected up to 6 h. The elimination rate constant was 0.35 +/- 0.02 /h and elimination half-life was 2.04 +/- 0.14 h. The apparent volume of distribution (Vd(area)) and total body clearance (ClB) were 1.21 +/- 0.15 l/kg and 0.41 +/- 0.03 l/kg/h, respectively. To maintain a minimum therapeutic concentration of 1 microgram/kg, a satisfactory dosage regimen of cefriaxone in febrile buffalo calves is 19 mg/kg followed by 18 mg/kg at 8 h intervals.
Animals ; Anti-Bacterial Agents/administration&dosage/*pharmacokinetics ; Area Under Curve ; Buffaloes/*metabolism ; Ceftriaxone/administration&dosage/*pharmacokinetics ; Drug Administration Schedule/veterinary ; Fever/drug therapy/*metabolism/*veterinary ; Half-Life ; Lipopolysaccharides/administration&dosage ; Male ; Metabolic Clearance Rate

OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.

METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.

RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).

CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.

Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology

BACKGROUND/AIMS: It is well known that alcohol enhances the toxicity of CCl4. We tried to establish an alcoholic liver cirrhosis model by administration of alcohol and CCl4 to rats. We also wanted to know the hepatoprotective effect of low doses of lipopolysaccharide(LPS) in this animal model. METHODS: Of 20 female adult rats, 8 were ingested with alcohol ad libitum(group 1) Another 6 were ingested with 10% alcohol and 50% 1mL/kg CCl4 intragastrically by Sonde twice a week(group 2) The remaining 6 were ingested with 10% alcohol, CCl4, and 0.1mg/kg LPS intraperitoneally twice a week(group 3) The fibrosis was evaluated semiquantitatively on a scale of 0(none) to 3(cirrhosis). RESULTS: 1) After 10 weks, septal fibrosis or cirrhosis was produced in 9 out of 12 rats in groups 2 and 3 but there was no fibrotic change in group 1. 2) There was no significant difference in pathological grading between groups 2 and 3. CONCLUSIONS: Hepatic fibrosis or cirrhosis can be sufficiently induced by alcohol and repetitive CCl4 ingestion for 10 weeks. We can not prove the hepatoprotective effect of low dose LPS by semiquantitative evaluation of pathological grading.
Animals ; Carbon Tetrachloride Poisoning/*complications ; English Abstract ; Ethanol/*toxicity ; Female ; Lipopolysaccharides/*administration & dosage ; Liver/pathology ; Liver Cirrhosis, Alcoholic/pathology/*prevention & control ; Rats ; Rats, Sprague-Dawley

PURPOSE: A bacterial extract consisting of immunostimulating components derived from 18 Escherichia coli strains has been used for the prophylaxis of recurrent cystitis. To evaluate the prophylactic effect of the E. coli extract, the cytokine levels of bladder tissue were measured after oral administration, and the bladder inflammation analyzed by histopathologic examination in a mouse model of lipopolysaccharide(LPS)-induced cystitis. MATERIALS AND METHODS: After 10-days of E. coli extract administration, the cytokine [interleukin-6(IL-6), IL-10, monocyte chemoattractant protein-1(MCP-1), interferon-gammaIFN-gamma, Tumor necrosis factor-alphaTNF-alpha, IL- 12p70] levels in the bladder of female Blab/C mice were determined using a cytometric bead array. The bladder macrophage inflammatory protein-2 (MIP-2) level was also measured using a sandwich enzyme immunoassay. After immunization with the E. coli extract, E. coli LPS was instilled into the bladders. Twenty-four hours later, the mice were sacrificed and the inflammation of the bladder quantified using the bladder inflammatory index (BII). RESULTS: Significant increases of the IL-6 and IFN-gammalevels in bladder tissue were observed after E. coli extract treatment. Secretions of the other cytokines were not stimulated by the E. coli extract. Bladders instilled with LPS showed high inflammation scores for edema, leukocyte infiltration, and hemorrhage in the saline treated control mice. In contrast, the E. coli extract treated mice exhibited mild inflammation of their bladders, with significant reduction in their BII scores compared to controls. CONCLUSIONS: These results demonstrate that immunization using oral treatment of E. coli extract provides protection against the inflammatory responses in a mouse model of LPS-induced cystitis.
Administration, Oral ; Animals ; Cystitis* ; Cytokines ; Edema ; Escherichia coli* ; Escherichia* ; Female ; Hemorrhage ; Humans ; Immunization ; Immunoenzyme Techniques ; Inflammation ; Interleukin-10 ; Interleukin-6 ; Leukocytes ; Lipopolysaccharides ; Macrophages ; Mice* ; Monocytes ; Necrosis ; Urinary Bladder

OBJECTIVETo observe the protection of carbon monoxide (CO) inhalation on lipopolysaccharide (LPS)-induced rat multiple organ injury.

METHODSSprague-Dawley rats with multiple organ injury induced by 5 mg/kg LPS intravenous injection were exposed to room air or 2. 5 x 10(-4) (V/V) CO for 3 hours. The lung and intestine tissues of rats were harvested to measure the expression of heme oxygenase-1 (HO-1) with reverse transcription-polymerase chain reaction, the levels of pulmonary tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and intestinal platelet activator factor (PAF), intercellular adhesion molecule-1 (ICAM-1) with enzyme-linked immunosorbent assay, the content of maleic dialdehyde (MDA) and the activity of myeloperoxidase (MPO) with chemical method, the cell apoptosis rate with flow cytometry, and the pathological changes with light microscope.

RESULTSCO inhalation obviously up-regulated the expression of HO-1 in lung (5.43 +/- 0.92) and intestine (6.29 +/- 1.56) in LPS + CO group compared with (3.08 +/- 0.82) and (3.97 +/- 1.16) in LPS group (both P < 0.05). The levels of TNF-alpha, IL-6 in lung and PAF, ICAM-1 in intestine of LPS + CO group were 0.91 +/- 0.25, 0.64 +/- 0.05, 1.19 +/- 0.52, and 1.83 +/- 0.35 pg/mg, respectively, significantly lower than the corresponding values in LPS group (1.48 +/- 0.23, 1.16 +/- 0.26, 1.84 +/- 0.73, and 3.48 +/- 0.36 pg/mg, all P < 0.05). The levels of MDA, MPO, and cell apoptosis rate in lung and intestine of LPS + CO group were 1.02 +/- 0.23 nmol/mg, 1.74 +/- 0.17 nmol/mg, 7.18 +/- 1.62 U/mg, 6.30 +/- 0.97 U/mg, 1.60% +/- 0.34%, and 30. 56% +/- 6.33%, respectively, significantly lower than the corresponding values in LPS group (1.27 +/- 0.33 nmol/mg, 2.75 +/- 0.39 nmol/mg, 8.16 +/- 1.49 U/mg, 7.72 +/- 1.07 U/mg, 3.18% +/- 0.51%, and 41.52% +/- 3.36%, all P < 0.05). In addition, injury of lung and intestine induced by LPS was attenuated at presence of CO inhalation.

CONCLUSIONCO inhalation protects rat lung and intestine from LPS-induced injury via anti-oxidantion, anti-inflammation, anti-apoptosis, and up-regulation of HO-1 expression.

Animals ; Base Sequence ; Carbon Monoxide ; administration & dosage ; DNA Primers ; Inhalation Exposure ; Lipopolysaccharides ; toxicity ; Male ; Multiple Organ Failure ; chemically induced ; Rats ; Rats, Sprague-Dawley

Tripterygium glycosides liposome(TPGL) were prepared by thin film-dispersion method, which were optimized accor-ding to their morphological structures, average particle size and encapsulation rate. The measured particle size was(137.39±2.28) nm, and the encapsulation rate was 88.33%±1.82%. The mouse model of central nervous system inflammation was established by stereotaxic injection of lipopolysaccharide(LPS). TPGL and tripterygium glycosides(TPG) were administered intranasally for 21 days. The effects of intranasal administration of TPG and TPGL on behavioral cognitive impairment of mice due to LPS-induced central ner-vous system inflammation were estimated by animal behavioral tests, hematoxylin-eosin(HE) staining of hippocampus, real-time quantitative polymerase chain reaction(RT-qPCR) and immunofluorescence. Compared with TPG, TPGL caused less damage to the nasal mucosa, olfactory bulb, liver and kidney of mice administered intranasally. The behavioral performance of treated mice was significantly improved in water maze, Y maze and nesting experiment. Neuronal cell damage was reduced, and the expression levels of inflammation and apoptosis related genes [tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), BCL2-associated X(Bax), etc.] and glial activation markers [ionized calcium binding adaptor molecule 1(IBA1) and glial fibrillary acidic protein(GFAP)] were decreased. These results indicated that liposome technique combined with nasal delivery alleviated the toxic side effects of TPG, and also significantly ameliorated the cognitive impairment of mice induced by central nervous system inflammation.
Mice ; Animals ; Tripterygium ; Liposomes ; Glycosides/therapeutic use* ; Administration, Intranasal ; Lipopolysaccharides ; Central Nervous System ; Cognitive Dysfunction/drug therapy* ; Inflammation/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Cardiac Glycosides

Carbon monoxide (CO), a metabolite of heme catalysis by heme oxygenase (HO), has been proposed to have anti-oxidative, anti-inflammatory and anti-apoptotic functions. Lipopolysaccharide (LPS)-induced lung injury (LI) is characterized by oxidative stress, inflammatory reaction and excessive pulmonary cell apoptosis. So we supposed that CO might have protection against LI. LI in rats was induced by intravenous injection of LPS (5 mg/kg). To observe the effect of CO inhalation, LI rats were exposed to 2.5 x 10(-4) (V/V) CO for 3 h. CO-induced changes of lung oxidative stress parameters, inflammatory cytokines, cell apoptosis, HO-1 expression and histology were examined. Results revealed that expressions of the tumor necrosis factor-alpha (TNF-alpha) and interlukin-6 (IL-6), activities of maleic dialdehyde (MDA) and myeloperoxidase (MPO), and cell apoptosis in LPS injection + CO inhalation group were (0.91+/-0.25) pg/mg protein, (0.64+/-0.05) pg/mg protein, (1.02+/-0.23) nmol/mg protein, (7.18+/-1.62) U/mg protein and (1.60+/-0.34)%, respectively, significantly lower than the corresponding values in LI group [(1.48+/-0.23) pg/mg protein, (1.16+/-0.26) pg/mg protein, (1.27+/-0.33) nmol/mg protein, (8.16+/-1.49) U/mg protein and (3.18+/-0.51) %, P<0.05]. Moreover, CO inhalation obviously increased the expressions of HO-1 and interlukin-10 (IL-10) and activity of superoxide dismutase (SOD) [(5.43+/-0.92), (0.26+/-0.07) pg/mg protein and (60.09+/-10.21) U/mg protein in LPS injection + CO inhalation group vs (3.08+/-0.82), (0.15+/-0.03) pg/mg protein and (50.98+/-6.88) U/mg protein in LI group, P<0.05]. LI was attenuated by CO inhalation. Our study demonstrates that inhalation of low concentration of CO protects lung against LPS-induced injury via anti-oxidant, anti-inflammation, anti-apoptosis and up-regulation of HO-1 expression.
Administration, Inhalation ; Animals ; Apoptosis ; drug effects ; Carbon Monoxide ; administration & dosage ; Carboxyhemoglobin ; analysis ; Cytokines ; biosynthesis ; Heme Oxygenase-1 ; genetics ; Lipopolysaccharides ; toxicity ; Lung ; drug effects ; metabolism ; pathology ; Male ; Oxidative Stress ; drug effects ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of berberine hydrochloride on the secretion of monocyte chemoattractant protein-1 (MCP-1) from human periodontal ligament cells (PDLC) in vitro culture.

METHODSPeriodontal ligament was isolated from extracted human premolars, and PDLC were cultured in vitro. PDLC were divided into two groups, lipopolysaccharide (LPS) group and non-lipopolysaccharide (NLPS) group. Then in accordance with the final concentrations of berberine hydrochloride in cells culture medium (0, 0.01, 0.02, 0.03 g/L), the groups were subdivided into LPS and NLPS control group, LPS1 and NLPS1 group, LPS2 and NLPS2 group, LPS3 and NLPS3 group. Cellular concentration of MCP-1 of each group was determined by enzyme-linked immunosorbent assay (ELISA). The data were statistically analyzed.

RESULTSThe MCP-1 contents were not significantly different between the groups of NLPS1, NLPS2 and NLPS3 [(11.33 ± 0.16), (11.45 ± 0.53), (11.25 ± 0.14) ng/L, respectively] and the NLPS control group [(11.32 ± 0.35) ng/L] (P = 0.692, 0.568, 0.524). MCP-1 contents in the groups of LPS1, LPS2 and LPS3 [respectively (38.14 ± 5.34), (34.15 ± 3.36), (26.13 ± 2.12) ng/L] were significantly lower than in LPS control group [(58.42 ± 1.52) ng/L], P = 0.000, 0.000, P = 0.000.

CONCLUSIONSThe inhibitory effect of berberine hydrochloride on the activities of MCP-1 from PDLC was more significant when PDLC were stimulated with LPS and in a concentration-dependent manner.

Adolescent ; Anti-Inflammatory Agents ; administration & dosage ; pharmacology ; Berberine ; administration & dosage ; pharmacology ; Bicuspid ; cytology ; Cells, Cultured ; Chemokine CCL2 ; secretion ; Child ; Dose-Response Relationship, Drug ; Humans ; Lipopolysaccharides ; pharmacology ; Periodontal Ligament ; cytology ; metabolism

OBJECTIVETo investigate the effect of hydrogen inhalation on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the underlying molecular mechanisms.

METHODSThirty-two male SD rats were randomly divided into 4 groups (n=8), namely the normal saline group (SA), saline with 2% hydrogen gas inhalation group (SH group), ALI group, and ALI with hydrogen inhalation group (LH group). In the two ALI groups, ALI was induced by intraperitoneal injection of 15 mg/kg LPS. Treatments with inhalation of 2% hydrogen gas for 6 h was administered after the injection of LPS or saline. The concentrations of tumor necrosis factor-α (TNF-α) in the lung tissue and serum were examined with ELISA. The expression of p38 MAPK in the lung tissue was detected by Western blotting..

RESULTSHydrogen inhalation decreased the expression of p-p38 MAPK in the lung tissue, and significantly reduced TNF-α content in the lung tissue and serum of rats with ALI.

CONCLUSIONHydrogen inhalation can decrease the expression of TNF-α in the lung tissue and serum, and this effect may be related with reduced p38 MAPK expression and inhibition of p38 MAPK activation.

Acute Lung Injury ; blood ; chemically induced ; metabolism ; Administration, Inhalation ; Animals ; Hydrogen ; administration & dosage ; Lipopolysaccharides ; Lung ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood ; metabolism ; p38 Mitogen-Activated Protein Kinases ; blood ; metabolism