OBJECTIVE: To investigate the effects of inflammation cytokines, (FK506) and cyclosporine (CSA) on albumin secretion, and the effects of FK506 and CSA on the IL-6 induced suppression of albumin synthesis in cultured human hepatocytes. METHODS: Human hepatoma cell lines (HepG2 cells) were separately cultured with IL-6, IL-2 and IL-10 (0 approximately 10 microg/L) and FK506, CSA (0 approximately 100 microg/L) for 48 h. In another experiment, HepG2 cells were stimulated with different doses of FK506 and CSA (0 approximately 10 microg/L) in the presence of IL-6 (5 microg/L) for 48 h. Albumin levels in the supernatant of all groups were measured by radioimmunoassay (RIA). The concentration of LDH secreted by cells stimulated with FK506 and CSA were detected with spectrophotometry. RESULTS: For cultured HepG2 cells, IL-6 significantly decreased albumin levels in a dose-dependent manner (P <0.01), and the maximal inhibition occurred at 5 microg/L. CSA mildly decreased albumin levels and a significant reduction in albumin production was first visible at 10 microg/ L (P <0.05). In contrast, IL-2, IL-10 and FK506 did not significantly influence albumin pro- duction (P > 0.05). FK506 obviously decreased LDH levels in the supernatant (P < 0.05) and attenuated IL-6 induced suppression of albumin synthesis (P < 0.01). But CSA slightly increased LDH concentration and could not block the IL-6 induced decrease of albumin synthesis (P > 0.05). CONCLUSION IL-6 but not IL-2 and IL-10 suppressed the production of hepatic albumin in vitro. FK506 protected against the suppression of hepatic albumin synthesis caused by IL-6, suggesting its potential role in improving hypoalbuminaemia in immune glomerulonephritis.
Albumins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclosporine ; pharmacology ; Hepatocytes ; physiology ; Humans ; Interleukin-10 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukin-6 ; pharmacology ; Liver Neoplasms ; metabolism ; pathology ; Tacrolimus ; pharmacology ; Tumor Cells, Cultured

Cell Survival ; drug effects ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; physiology ; Humans ; Immunophenotyping ; Interleukin-10 ; pharmacology ; Interleukin-12 ; genetics ; secretion ; Interleukin-12 Subunit p35 ; Interleukin-12 Subunit p40 ; Protein Subunits ; genetics ; secretion ; RNA, Messenger ; analysis

PURPOSE: Hyperoxia has the chief biological effect of cell death. We have previously reported that cathepsin B (CB) is related to fetal alveolar type II cell (FATIIC) death and pretreatment of recombinant IL-10 (rIL-10) attenuates type II cell death during 65%-hyperoixa. In this study, we investigated what kinds of changes of CB expression are induced in FATIICs at different concentrations of hyperoxia (65%- and 85%-hyperoxia) and whether pretreatment with rIL-10 reduces the expression of CB in FATIICs during hyperoxia. MATERIALS AND METHODS: Isolated embryonic day 19 fetal rat alveolar type II cells were cultured and exposed to 65%- and 85%-hyperoxia for 12 h and 24 h. Cells in room air were used as controls. Cytotoxicity was assessed by lactate dehydrogenase (LDH) released into the supernatant. Expression of CB was analyzed by fluorescence-based assay upon cell lysis and western blotting, and LDH-release was re-analyzed after preincubation of cathepsin B-inhibitor (CBI). IL-10 production was analyzed by ELISA, and LDH-release was re-assessed after preincubation with rIL-10 and CB expression was re-analyzed by western blotting and real-time PCR. RESULTS: LDH-release and CB expression in FATIICs were enhanced significantly in an oxygen-concentration-dependent manner during hyperoxia, whereas caspase-3 was not activated. Preincubation of FATIICs with CBI significantly reduced LDH-release during hyperoxia. IL-10-release decreased in an oxygen-concentration-dependent fashion, and preincubation of the cells with rIL-10 significantly reduced cellular necrosis and expression of CB in FATIICs which were exposed to 65%- and 85%-hyperoxia. CONCLUSION: Our study suggests that CB is enhanced in an oxygen-concentration-dependent manner, and IL-10 has an inhibitory effect on CB expression in FATIICs during hyperoxia.
Animals ; Cathepsin B/*genetics/metabolism ; *Down-Regulation ; Gene Expression Regulation ; Hyperoxia/*genetics ; Interleukin-10/*pharmacology/physiology ; L-Lactate Dehydrogenase/metabolism ; Necrosis/chemically induced ; Oxygen/metabolism ; Rats

BACKGROUNDWe investigated the role of 1,25-dihydroxyvitamin D3(1,25-(OH)2D3) in preventing allograft from acute rejection following orthotopic liver transplantation.

METHODSA rat orthotopic liver transplantation model was used in this study. SD-Wistar rats served as a high responder strain combination. Recipients were subjected to administration of 1,25-(OH)2D3 at dosages ranging from 0.25 microg.kg(-1).d(-1) to 2.5 microg.kg(-1).d(-1). Survival after transplantation as well as pathological rejection grades and IFN-gamma mRNA, IL-10 mRNA transcription intragraft on day 7, and day 30 post-transplantation were observed.

RESULTSAfter recipients were treated with 1,25(OH)2D3 at dosages of 0.5 microg.kg(-1).d(-1) or 1.0 microg.kg(-1).d(-1), survivals of recipients were prolonged. Ninety-five percent confidence intervals of survival were 46 - 87 days and 69 - 102 days (both P = 0.0005 vs control group), respectively. On day seven post-transplantation, relative levels of IFN-gamma mRNA transcription were 0.59 +/- 0.12 and 0.49 +/- 0.16, which was higher than the control group (P = 0.005, P = 0.003, respectively). Relative levels of IL-10 mRNA transcription were 0.83 +/- 0.09 and 0.76 +/- 0.09, which was lower than the control group (P = 0.002, P = 0.003, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and on day thirty post-transplantation were 1.5 and 2.0 in comparison with the CsA-treated group (P = 0.178, P = 0.171, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and day thirty post-transplantation were 1.5 and 1.5 in comparison with CsA-treated group (P = 0.350, P = 0.693, respectively).

CONCLUSIONAfter each recipient was treated with 1,25-(OH)2D3 at a dosage of (0.5 - 1.0) microg.kg(-1).d(-1), transcription of cytokine intragraft was accommodated effectively and deviated to Th2 type, resulting in alleviation of acute rejection. 1,25-(OH)2D3 can prolong survival of recipient after orthotopic liver transplantation.

Animals ; Calcitriol ; pharmacology ; physiology ; Graft Rejection ; prevention & control ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Liver Transplantation ; mortality ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transcription, Genetic

Chemokine KC has been considered to be a murine homologue of human GRO/MGSA and was identified as chemoattractant for monocytes and neutrophils. This study examined the expression of KC mRNA in thioglycollate-elicited mouse peritoneal macrophages that were stimulated in vitro with Candida albicans (CA). Also examined were the inhibitory effects of IL-10 on the CA-induced expression of KC gene by Northern blot analysis. CA was found to induce chemokine gene expression in a gene-specific manner, CXC chemokine IP-10 mRNA expression was not detected in CA-stimulated macrophages. Maximum KC mRNA expression was observed approximately 2 hr after adding CA. The inhibitory action of IL-10 to CA-induced KC mRNA expression on mouse peritoneal macrophages was independent on concentration and stimulation time of IL-10 and was observed approximately one hour after adding IL-10 and CA simultaneously. IL-10 produced a decrease in the stability of KC mRNA, and CA-stimulated macrophages with cycloheximide blocked the suppressive effect of IL-10. These results suggest that CA also induces chemokine KC from macrophages, and IL-10 acts to destabilize CA-induced KC mRNA and de novo synthesis of an intermediate protein is a part of the IL-10 suppressive mechanism.
Animal ; Blotting, Northern ; Candida albicans/metabolism* ; Cells, Cultured ; Chemotactic Factors/genetics* ; Dactinomycin/pharmacology ; Dose-Response Relationship, Drug ; Gene Expression Regulation/drug effects* ; Growth Substances/genetics* ; Interleukin-10/pharmacology* ; Interleukin-10/metabolism ; Macrophages/physiology* ; Mice ; Mice, Inbred BALB C ; Nucleic Acid Synthesis Inhibitors/pharmacology ; RNA, Messenger/metabolism ; RNA, Messenger/drug effects

OBJECTIVETo investigate the possible roles of adenosine and the cytokines TNF-alpha and IL-10 in the pathogenesis of acute bacterial prostatitis (ABP) in rats.

METHODSForty-eight male Wistar rats were randomly divided into groups A (ABP), B (ABP + theophylline intervention), C (sham) and D (blank control). ABP models were established by injecting Escherichia coli 0157 into the prostate, and those in group B were treated by intraperitoneal injection of theophylline immediately after modeling. At 4 and 14 days, the prostate tissues of the rats were collected for detection of the expressions of TNF-alpha and IL-10 by immunohistochemistry and the concentration of adenosine by high-performance liquid chromatography.

RESULTSAt 4 and 14 days, the concentrations of adenosine were significantly higher in group A ([48.38 +/- 17.27] and [26.54 +/- 11.22] microg/g) than in C ([0.45 +/- 0.25] and [0.46 +/- 0.29] microg/g) and D ([0.41 +/- 0.23] and [0.43 +/- 0.27] microg/g) (P < 0.05), and so were the expressions of TNF-alpha in A (0.23 +/- 0.08 and 0.21 +/- 0.03) than in C (0.07 +/- 0.03 and 0.07 +/- 0.01) and D (0.07 +/- 0.06 and 0.07 +/- 0.06) (P < 0.05), and those of IL-10 in A (0.13 +/- 0.03 and 0.25 +/- 0.01) than in C (0.07 +/- 0.03 and 0.07 +/- 0.03) and D (0.07 +/- 0.01 and 0.07 +/- 0.02) (P < 0.05). Compared with group A, the rats in group B showed significant increases at 4 and 14 days in the severity of inflammation, concentration of adenosine ([86.64 +/- 32.87] and [51.17 +/- 22.96] microg/g, P < 0.05) and expression of TNF-alpha (0.37 +/- 0.08 and 0.32 +/- 0.06, P < 0.05), but exhibited no remarkable difference in the expression of IL-10 (0.12 +/- 0.06 and 0.15 +/- 0.06, P > 0.05).

CONCLUSIONAdenosine may affect the progression of inflammation by regulating the expressions of the cytokines TNF-alpha and IL-10 in ABP rats through the adenosine receptor signaling pathway.

Adenosine ; physiology ; Animals ; Escherichia coli O157 ; Interleukin-10 ; metabolism ; Male ; Prostate ; drug effects ; metabolism ; Prostatitis ; metabolism ; microbiology ; Random Allocation ; Rats ; Rats, Wistar ; Theophylline ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the impact of AG490, a cytokine signaling inhibitor, on cytokine signaling pathway with phosphorylation levels of Janus kinase 2 (Jak2) and singal transducers and activators of transcription 3 (Stat3), and liver pro-/anti-inflammatory cytokine expressions.

METHODSRats were divided into two groups after surgery: control group, without treatment; AG490 group, with AG490 (1 mg.kg(-1).12 h(-1)) administration intraperitoneally, immediately and through 36 hs after the operation. Western blotting was used to detect the levels of phosphorylated Jak2 and Stat3. Semi-quantitative RT-PCR was employed to examine Interleukin-6 (IL-6) and Interleukin-10 (IL-10) expression.

RESULTSAt 8 h and 12 h post-operatively, the phosphorylation levels of Jak2 and Stat3 were significantly inhibited in the AG490 group when compared with the control group. The DNA levels of IL-6 in the liver of the AG490 group rat at the same time points were also decreased, whereas IL-10 levels markedly increased. These changes made the ratio of IL-6/IL-10 dropped significantly.

CONCLUSIONSAG490 ameliorates the overwhelming inflammatory response via a mechanism of blocking cytokine signaling transduction and consequently suppresses the ratio of pro-/anti-inflammatory cytokine expression, which exerts potential clinical implications of use of anti-inflammatory agents in hepatic surgery.

Animals ; Cytokines ; biosynthesis ; DNA-Binding Proteins ; metabolism ; Hepatectomy ; methods ; Interleukin-10 ; biosynthesis ; Interleukin-6 ; biosynthesis ; Janus Kinase 2 ; Liver ; drug effects ; physiology ; Male ; Protein-Tyrosine Kinases ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; STAT3 Transcription Factor ; Signal Transduction ; drug effects ; physiology ; Trans-Activators ; metabolism ; Tyrphostins ; pharmacology

Cisplatin, a major anti-neoplastic drug, is known to be nephrotoxic and inflammation-inducing. A peroxisome proliferator-activated receptor gamma agonist, regulating lipid metabolism, has known to have anti-inflammatory effect, but the protection mechanisms in various kidney injuries are not fully understood. The purpose of this study was to examine the reno-protective effect of rosiglitazone on cisplatin nephrotoxicity in mice focusing on inflammation and apoptosis. Male BALB/c mice were pretreated with rosiglitazone (10 mg/kg) or vehicle through daily intraperitoneal injection for 3 days and then were given a single injection of cisplatin (20 mg/kg). Cisplatin induced a significant rise in blood urea nitrogen and creatinine levels, and tubular cell damage with marked tissue inflammation. Tissue cytokines and chemokines measured by a cytometric bead array showed increased TNF-alpha, IL-6, MCP-1, and IFN-gamma levels, while IL-10, an anti-inflammatory cytokine, was significantly decreased by cisplatin treatment. However, rosiglitazone pretreatment substantially reversed the depressed IL-10 level with simultaneous suppression of proinflammatory cytokines and chemokines. This tissue cytokine and chemokine milieu was associated with marked attenuation of kidney injury elicited by cisplatin. These findings suggest that the rosiglitazone-mediated renoprotective effect in cisplatin nephrotoxicity of mice is partially mediated by upregulation of anti-inflammatory IL-10 production.
Acute Kidney Injury/*chemically induced ; Animals ; Apoptosis/physiology ; Caspases/metabolism ; Chemokines/metabolism ; Cisplatin/*toxicity ; Cytokines/metabolism ; Enzyme Activation ; Hypoglycemic Agents/*pharmacology ; Interleukin-10/*metabolism/pharmacology ; Kidney/*drug effects/metabolism/pathology ; Male ; Mice ; Mice, Inbred BALB C ; PPAR gamma/metabolism ; Thiazolidinediones/*pharmacology

OBJECTIVETo investigate the effect of interleukin-10 (IL-10) on the expression of transforming growth factor-beta(1) (TGFbeta(1)) and platelet-derived growth factor (PDGF) in hepatic stellate cells (HSC) during liver injury.

METHODSThe adenovirus vector (the titer was 1 x 10(7) efu/ml) encoded IL-10 gene was used to transfect the rat via the vein of caudal. At the same time, CCl(4) was injected into rat by a hypodermic injection. These processes went on twice a week. After eight weeks, the liver were perfused with collagenase IV and purified by density gradient centrifugation with Nycodenz for separate HSC. The level of IL-10 was measured by ELISA method; The expression of PDGF and TGFbeta(1) in HSC was detected by semi-quantitative RT-PCR and Western-blot methods.

RESULTSThe level of IL-10 in therapy group (adenovirus vector encoding IL-10 gene group) was higher than that in non-therapy group (adenovirus vector without IL-10 gene and PBS group); The expression of TGFbeta(1) mRNA, TGFbeta(1) protein and PDGF mRNA, PDGF protein in therapy group were significantly lower than that in non-therapy group (P < 0.05).

CONCLUSIONDownregulating the TGFbeta(1) and PDGF expression could be the passageway by which IL-10 alleviate the degree of proliferation and activation in hepatic stellate cells.

Animals ; Down-Regulation ; drug effects ; Genetic Therapy ; Hepatocytes ; drug effects ; physiology ; Interleukin-10 ; pharmacology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; therapy ; Male ; Platelet-Derived Growth Factor ; biosynthesis ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; drug effects ; physiology ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; Transforming Growth Factor beta1

OBJECTIVETo study the protective function and pathophysiology of cystathionine gamma-lyase (CSE)/hydrogen sulfide (H(2)S) system in hepatic ischemia-reperfusion injury (HIRI) in rats.

METHODSWistar rats were randomly distributed into sham group (n = 18), ischemia-reperfusion (IR) group (n = 18), IR + NaHS group (n = 18) and IR + DL-propargylglycine (PAG) group (n = 18). The hepatic IR model was established by Pringle's hepatic vascular occlusion. At each of the indicated time points (1, 3 and 6 hours after IR), the serum levels of H(2)S and the hepatic CSE activity were measured. The serum levels of inflammatory factors, including TNF-α, IL-10 were determined by ELISA methods. The expression of apoptotic protein, TNF-α, in liver tissue was tested by Western blot assay, cell apoptosis was examined by TUNEL and the histological changes were examined in each group.

RESULTSThe serum levels of H(2)S and CSE activity were significantly increased in group IR compared with group sham at all indicated time points (P < 0.05). The serum level of inflammatory factors (P < 0.01) and the hepatic expression of TNF-α protein (P < 0.05) were elevated obviously in group IR than that in group sham. Administration of NaHS could reduce the production of inflammatory factors in serum (P < 0.01), inhibit hepatic protein expression of TNF-α (P < 0.05) and attenuate the liver histological scores of IR injury (P < 0.05), whereas PAG aggravated them.

CONCLUSIONThe endogenous CSE/H(2)S system maybe involved in the pathogenesis of hepatic IR injury, which suggests that CSE/H(2)S system can protect liver from IR injury in rats by intervening in inflammatory reaction, attenuating the injury severity and inhibiting expression of apoptotic protein TNF-α.

Animals ; Apoptosis ; drug effects ; Cystathionine gamma-Lyase ; blood ; physiology ; Disease Models, Animal ; Hydrogen Sulfide ; blood ; Interleukin-10 ; blood ; Liver ; blood supply ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology ; physiopathology ; prevention & control ; Sulfides ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism