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[ABSTRACT]AIM:ToobservehowfarnesoidXreceptor(FXR)functionedinconcanavalinA(ConA)-induced hepatitis (CIH) and the regulation of FXR-thyrotropin embryonic factor (TEF) pathway.METHODS:C57BL/6 mice were injected with Con A to induce hepatitis .The expression of FXR and TEF in the liver specimens was determined by qRT-PCR and Western blotting .The concentrations of serum ALT/AST and inflammatory cytokines IFN-γ, TNF-α, IL-4 and IL-2 in the blood samples were tested after Con A injection .RESULTS:FXR was down-regulated in CIH mice .TEF was up-regula-ted when FXR was activated by chenodeoxycholic acid (CDCA).Activation of FXR reduced the levels of aminotransferases and inflammatory cytokines IFN-γ, TNF-α, IL-4 and IL-2 in the CIH mice induced by Con A injection .CONCLUSION:FXR activation attenuates CIH mouse liver injury and reduces inflammatory cytokines .FXR activation results in TEF up-regu-lation.The FXR-TEF pathway may play a protective role in autoimmune hepatitis .
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<p><b>OBJECTIVE</b>To determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).</p><p><b>METHODS</b>Peripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot.</p><p><b>RESULTS</b>Levels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA.</p><p><b>CONCLUSION</b>IL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.</p>
الموضوعات
Adolescent , Adult , Humans , Middle Aged , Cells, Cultured , DNA-Binding Proteins , Metabolism , Interleukin-12 , Blood , Genetics , Leukocytes, Mononuclear , Metabolism , Lupus Erythematosus, Systemic , Metabolism , Phosphorylation , RNA, Messenger , STAT3 Transcription Factor , STAT4 Transcription Factor , Trans-Activators , Metabolismالملخص
【Objective】To observe the expression of PI3-K phosphorylated products and elucidate the correlation between PI3-K phosphorylated products and Th2 cytokine in peripheral blood mononuclear cell (PBMC).【Methods】14 patients with active lupus nephritis and 12 controls were selected,PI3-K phosphorylated products were detected by immunoprecipitation and Western blotting,RT-PCR was used to observe interleukin-6 mRNA and interleukin-10 mRNA expression.【Results】In either spontaneous condition or stimulated by anti-CD3 antibody,the expression of PI3-K phosphorylated products in patients with active lupus nephritis were higher than those of the controls(1.14±0.23 vs 0.46±(0.12，P=0.023;2.09±0.63 vs 0.65±0.14，P=0.016).The expression of PI3-K phosphorylated products in active lupus nephritis showed a positive correlation with interleukin-6 mRNA and interleukin-10 mRNA (r=0.652,P=0.008;r=0.718,P=0.007).PY294002,one of specific inhibitor of PI3-K,inhibited significantly the expression of interleukin-6 mRNA(2.32±0.51 vs 0.57±0.15,P=0.009) and interleukin-10 mRNA (1.71±0.33 vs 0.67±0.11,P=0.006) in stimulated PBMC in active lupus nephritis.【Conclusion】PI3-K can involve in the pathogenesis of lupus nephritis by inducing the overexpression of interleukin-6 and interleukin-10.
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AIM:To investigate the regulatory role of nuclear factor κB (NF-κB) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1β.METHODS:Activation of NF-κB was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS:rhIL-1β could rapidly stimulate the activation of NF-κB in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-κB， could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1β.CONCLUSION:IL-6 expression induced by IL-1β may be regulated by NF-κB in MC, NF-κB may modulate the immune-inflammatory reaction in glomerular disease.
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Objective:To detect the activation of NF AT and AP 1 in peripheral blood mononuclear cells(PBMCs) in patients with systemic lupus erythematosus and investigate their relationship with serum ANA,anti dsDNA antibody,ESR,CRP and C 3.Methods:Activation of NF AT and AP 1 were detected by electrophoretic mobility shift assay(EMSA).Results:①Activation of NF AT in PBMC in active and inactive SLE patients was increased when compared with normal controls,respectively,and the activation of NF AT in active patients was higher than that of inactive ones.②Elevated activation of AP 1 in PBMC in active SLE patients was found as compared with normal controls,and there was no significant difference in activation of AP 1 between inactive patients and normal control.③Activation of NF AT in SLE patients with positive anti dsDNA antibody was higher than that of patients with negative anti dsDNA antibody,and there was not different positively in AP 1 activation between above two patient groups.④Activation of NF AT in PBMC was related positively with CRP and SLEDAI,respectively,but no correlation with ESR,ANA,C 3.There was no relation between the activation of AP 1 and above clinical parameters.Conclusion:The abnormal expression of NF AT and AP 1 indicates disturbed intracellular signaling in SLE,which is suggested that alternations in activation of transcription factors may contribute to the pathogenesis of SLE.Activation of NF AT may be used as a predicator of SLE disease activity.
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AIM: To examine whether Akt signal pathway proteins, including Akt, NF-?B and I?B?, are activated in kidney tissue of murine chronic graft-versus -host disease (GvHD) lupus nephritis in vivo , and whether prednisone suppres ses activation of them. METHODS: Akt activity and phosphorylated I?B? were detected by Weste rn-blot. Activation of NF-?B was detected by electropheretic mobilit y shift assay (EMSA). RESULTS: Activity of Akt, NF-?B and phosp horylated I?B ? were significantly increased in kidney tissue of murine chronic graft-versus -ho st disease (GvHD) in 8th week and 12th week after monocell injection, respective ly. However, they were no significant elevation in 16th week, when compared with controls. Prednisone treatment significantly prevented the increase in serum an ti-dsDNA antibody level, urinary protein excretion and glomerular cell prolif eration in GvHD mice, indicating the beneficial effects of prednisone on t his model. Prednisone also significantly suppressed the increase in the activities o f glomerular Akt, NF-?B and phosphorylated I?B?. CONCLUSION: T his study provides t he first evidence of marked increase in glomerular Akt-NF-?B signal pathway act ivities in murine chronic graft-versus-host disease lupus nephritis. The benefic ial effect of prednisone on this lupus nephritis model may be partially mediated by the suppression of abnormal Akt- NF-?B activation.
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AIM:To observe the expression of chemokine fractalkine,and its receptor,CX3CR1,in kidneys of lupus-prone BXSB mice,and their changes after treatment with prednisone. The role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was also discussed. METHODS:Twelve 12-week-old male BXSB mice were randomly divided into two groups,the prednisone treatment group (BXSB-prednisone group,n=6) and the experimental control group (BXSB group,n=6). Six male C57BL/6J mice at the same weeks of age served as a normal control group (C57BL/6J group). Both the C57BL/6J and the BXSB group of mice received a daily intragastric administration of 0.5 mL normal saline. The BXSB-prednisone group of mice was given a daily intragastric administration of prednisone (0.18 mg/20 g BW) dissolved in 0.5 mL normal saline. All treatments lasted for 10 weeks. The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of mice were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis respectively. The changes of laboratory index and the kidney histopathology of mice were also investigated. RESULTS:The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of BXSB mice were significantly higher than those in C57BL/6J mice. The expressions of fractalkine and CX3CR1 in BXSB-prednisone group of mice were much lower than those in BXSB group of mice,accompanied by the lower serum IgG,IgM and anti-dsDNA antibody levels as well as blood urea nitrogen,serum creatinine and urine protein. The glomerular immune complex deposition and the kidney histopathology were also significantly improved in BXSB-prednisone group of mice. CONCLUSION:These results indicate that fractalkine and CX3CR1 participate in the pathogenesis of lupus nephritis in BXSB mice,and the effect of glucocorticoids treatment may be attributed,in part,to its ability to inhibit the expression of fractalkine in kidney.
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AIM: To determine the expression of membrane-bound B lymphocyte stimulator((BLyS)) and its mRNA in peripheral blood mononuclear cells(PBMCs) from individuals with systemic lupus erythematosus(SLE),and to investigate the effect of dexamethasone on(BLyS) expression.METHODS: PBMCs were obtained from 25 individuals with SLE(mean age of 31.40?14.23) and 20 female healthy volunteers(mean age of 28.20?10.36).They were randomized into dexamethasone((1 ?mol/L)) group and media group.PBMCs were gathered at 0,6,12 and 24 h for(BLyS) mRNA assessment using reverse transcription-PCR(RT-PCR).PBMCs were also collected at 72 h for membrane-bound(BLyS) protein detection using flow cytometry(FACS) and direct immunofluorescence.RESULTS:(1) The expression of(BLyS) mRNA and membrane-bound protein were significantly higher in PBMCs from individuals with SLE than that in PBMCs from healthy controls(0.40?0.18 vs 0.27?0.20,P
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AIM: To investigate the role of CD134 (OX40) and NF-?B in the pathogenesis of lupus nephritis (LN). METHODS: Renal in situ CD134 and NF-?B expression were examined in 40 biopsy specimens from LN patients by immunohistochemistry and microwave-based immunohistochemistry, respectively. The relationship between expression of CD134 and NF-?B was analyzed. RESULTS: The expression of glomerular and tubular CD134 and NF-?B in LN were higher than that in normal control, especially in class Ⅳ LN, where there was intense staining of endothelial cell, distal tubules, and interstitial mononuclear cell. The CD134 expression of glomerular and tubular was closely related to NF-?B expression, respectively ( r=0.5542,P
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Objective To detect the activation of NF－? B signaling pathway in peripheral blood mononuclear cells(PBMCs) of patients with systemic lupus erythematosus(SLE) and investigate its clinical significance. Methods Activation of NF－? B was detected by electrophoretic mobility shift assay( EMSA), the expression of I? B? protein and its phosphorylated products were detected by Western blot. Anti-dsDNA antibody, IgG and IgM in supernatant of PBMC culture were tested by ELISA. Results (1)Elevated activation of NF－? B in PBMC of SLE patients was found when compared with the controls(P
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Objective To investigate the role of NF-?B in the pathogenesis of lupus nephritis (LN). Methods A microwave-based immunohistochemistry (APAAP) was used to detect the expression of NF-?B p65 and C-myc protein in renal tissue of LN and normal control. At the same time, the correlation between expression of NF-?B and renal injuries in LN was examined. Results The expression of NF-?B in LN was higher than that of normal control, the highest one was type Ⅳ LN. The number of positive cells expressing NF-?B in renal tissue of LN showed a strong correlation with C-myc protein expression, kidney activity of LN, histological changes and functional lesion in LN, respectively. Conclusions NF-?B may play an important role in the pathogenesis of LN, and the number of positive cells expressing NF-?B may be considered as a predictor for the kidney activity and progressive renal injury in LN.
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AIM: To investigate the effect of IL-4, CD40L on RANTES production in murine renal tubular epithelial cells (TEC). METHODS: TEC were obtained from mouse, expression of RANTES and CD40 on TEC were measured. RESULTS: (1) Activation of TEC with IL-4 resulted in significant increase in CD40 expression (P
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AIM: To explore the role of Akt/NF-?B pathway in immune-complexes-induced monocyte chemoattractant protein-1 (MCP-1) and colony stimulating factor-1 (CSF-1) expression in Mesangial Cells. METHODS: Primary murine glomerular mesangial cells were cultured in vitro and divided into control group, stimulation group and antisense, sense and mismatched oligodeoxynucleotide group. In control group, the cells were stimulated with monomeric IgG after treatment with 0.5% lipofectin for 8 h. In stimulation group, the cells, which had been treated with 0.5% lipofectin for 8 h, were stimulated with aggregated IgG. In antisense, sense and mismatched oligodeoxynucleotide group, being transduced antisense, sense and mismatched oligodeoxynucleotide respectively with 0.5% lipofectin 8 h, the cells were stimulated with AIgG. MCP-1 and CSF-1 in supernatant were deteced with ELISA. In addition, RT-PCR was used to determine MCP-1 and CSF-1 mRNA expression, and EMSA to investigated the activation of NF-?B. RESULTS: Mesangial cells cultured in vitro had a low level NF-?B activation and a low level constitutive expression of MCP-1 and CSF-1. Stimulated with AIgG, activation of NF-?B was markedly increased(0.35?0.06 vs 0.75?0.16, P
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AIM: To detect the association between the polymorphism of Fc receptor ? chain gene at position-29 in promoter and systemic lupus erythematosus(SLE). METHODS: The genotypes at position -29 in promoter of Fc receptor ? chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China. RESULTS: The frequencies of TT genotype(33.3%) and T allele (54 4%) at position -29 in patients with SLE were significantly higher than those in controls (17 9%, respectively), whereas, the frequencies of GG genotype (24 4%) and G allele (45 6%) in patients with SLE were remarkably lower than those in controls (31 4% and 57 1%, respectively) ( P 0 05) . CONCLUSION: Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.
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AIM: To investigate the regulatory role of nuclear factor ?B (NF-?B) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1?.METHODS: Activation of NF-?B was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS: rhIL-1? could rapidly stimulate the activation of NF-?B in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-?B, could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1?.CONCLUSION: IL-6 expression induced by IL-1? may be regulated by NF-?B in MC, NF-?B may modulate the immune-inflammatory reaction in glomerular disease.