ABSTRACT
<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>
Subject(s)
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 , MetabolismABSTRACT
Objective:To detect MIP 1? mRNA in kidney specimens in lupus nephritis(LN) patients and investigate its association with clinical and pathological features.Methods:MIP 1? mRNA were identified by in situ hybridization.Results:MIP 1? mRNA were detected in gromeluli,cortical tubular cells and interstitum of LN specimens wherase no MIP 1? mRNA was found in normal specimens.There were especially obvious MIP 1? expression in IV type LN specimens.MIP 1? expression in LN gromeruli,tubular cells and interstitium were correlated positively with LN histological activity index.Conclusion:There were enhancement of MIP 1? mRNA expression in LN kidney specimens,especially in IV type LN.MIP 1? expression in LN specimens were related with active inflammatory injury of glomeruli and tubulointerstitium.
ABSTRACT
Aim To investigate whether there is IL-12 signaling pathway through lck/P38/c-jun in splenic cells obtained from lupus mouse and its effect on splenic cells. Methods Mice with graft versus host disease were used as lupus nephritis model. Activity of Lck tyrosine kinase, p38 phosphorylation and mRNA expression of c-jun in splenic cells were determined by autoradiography, Western blot and Northern blot, respectively. Results There were higher levels of Lck activity, p38 phosphorylation and c-jun expression of IL-12-stimulated splenic cells from lupus model when compared with that observed in similarly treated splenic cells from normal control. The Lck activity and p38 phosphorylation were almost inhibited by Lck inhibitor PP1, on the other hand, p35 specific inhibitor SB203580 decreased phosphorylation of p38. In addition, expression of c-Jun was also inhibited by PP1 or SB203580 although splenic cells were stimulated with IL-12. Conclusion Aberrant murine splenic cell, IL-12-me-diated intracelluar signaling pathway through lck/p38/c-Jun were involved in immunologic damage.
ABSTRACT
【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.
ABSTRACT
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.
ABSTRACT
Objective:To investigate chemokine RANTES(Regulated upon Activation.Normal T cell Expressed and Secreted)expression in the chronic graft versus host disease(GVHD) murine lupus nephritis model and effect of the recipe of LangChuangFang on it.Methods:A microwave based immunohistochemistry(SP) and RT PCR were used to detect the expression of RANTES in the kidney of model,therapeutic group and normal control.At the same time,the correlation between expression of RANTES and renal injures in model was examined.Results:LangChuangFang protected renal function,which was coincided with amelioration of pathologic lesions especially tubulointersitial injury.This was associated with the amelioration of RANTES protein and mRNA expression in model.Conclusion:RANTES involved in the process of renal injury in vivo,LangChuangFang can treat lupus as immunosuppressive agent by decreasing RANTES production.
ABSTRACT
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.
ABSTRACT
AIM: To investigate the relationships between I?1 hs1,2 VNTR polymorphism and IgA nephropathy. METHODS: Four hundred and ninteen patients with IgA nephropathy and their first-degree relatives were recruited. Two hundred and one sex and age-matched normal Chinese Han volunteers were also recruited as controls. After extracting genomic DNA, the VNTR genotypes of I?1 hs1,2 region were determined by PCR and electrophoresis, and the results were analyzed by transmission disequilibrium test (TDT) and haplotype relative risk (HRR) in the families, and Chi-Square test in the case-control analysis. RESULTS: ① TDT analyses showed that B allele of the I?1 hs1,2 VNTR region was significantly more transmitted from heterozygous parents to patients than expected (101 Trios, ?2=6.818, P
ABSTRACT
AIM: To investigate the association of gene polymorphism at position 196 of tumor necrosis factor receptor Ⅱ (TNFRⅡ) with systemic lupus erythematosus (SLE) in Chinese, and establish recombinant retroviral vector to analyze the function of the TNFRⅡ 196M/R. METHODS: The genotype at position 196 of TNFRⅡ was determined by PCR-RFLP in 106 SLE patients and 119 healthy controls in china. Human TNFRⅡ196M cDNA were amplified by PCR and cloned into PMD18-T vector. Then, PMD18-TNFRⅡ196R was induced by site-directed mutagenesis. The recombinant T vector, PMD18-TNFRⅡ196M and PMD18-TNFRⅡ196R, were subcloned into retroviral vector PLXSN. Both normal and variant were transfected into rat mesangial cell. The effects of TNF? on production of sTNFRⅡ and IL-6 were study by ELISA. RESULTS: (1) The frequency of TNFRⅡ196R allele was significantly higher than those in controls (35.2% vs 14.3%, P
ABSTRACT
AIM: To study the effect of Bene Jones protein (BJP) from multiple myeloma(MM) patient and TGF-? 1 on cultured renal proximal tubular cell(PTC) proliferation. METHODS:[ 3H] TdR incorporation was used to study the effect of ?BJP and TGF-? 1 on cultured rat NRK.52E PTC proliferation,the expression of TGF-? 1 in the supernatant of PTC cultured with BJP was assessed with ELISA. RESULTS:① [ 3H] TdR incorporation of PTC was inhibited by BJP in a dose-dependent manner,when co-cultured with 100-800 ?mol/L BJP and 2.0 ?g/L TGF-? 1, the [ 3H] TdR incorporation was lower than that of BJP alone, especially when BJP≥400 ?mol/L; ②The expression of TGF-? 1 in the supernatant of PTC cultured with BJP was increased ,especially when BJP≥400 ?mol/L( P
ABSTRACT
AIM:To study the effects and mechanism of recombinant human defensin ?1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS:The influences of defensin ?1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase(MEK)inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin ?1,the phosphorylation of extracellular signal regulated kinase(ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS:Defensin ?1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h(P20 mg/L decreased cell proliferation.The cell proliferation induced by defensin ?1 was inhibited by U0126.Stimulation of the cells with defensin ?1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin ?1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h(P
ABSTRACT
To observe the role of parathyroid hormone PTH( 1 - 84) and platelet cytosolic free calcium I pt[ Ca2 + ] i in regulation of blood pressure on hemodialysis (HD) patients. Methods Fura-2 fluorometry and immunoradioassy werw used to detect PTH(1 - 84) and pttCa2+ ]i in 24 HD patients. Results The resting pt[Ca?+ ]i and PTH(1 - 84) increased in HD cases, and they were significantly higher in uremic patients with hypertension than ones without hypertension. The resting pt[Ca2+ ]i was correlated to PTH( 1 - 84) in HD cases. The multivariate stepwise regression analysis showed the PTH( 1 - 84) and pt[Ca2 + ]i are probably mains factors to affect the blood pressure in HD cases. Conclusion PTH(l - 84) and pt[Ca2+ ji are main factors affecting the blood pressure in uremic patients.
ABSTRACT
To investigate the pathogenic mechanism of interleukin 6(IL-6) in the tubulointerstitial lesions of patients with active lupus nephritis(LN). Methods Uninary IL-6 from 42 active LN cases and the renal tissues sections from 15 of them were examined by performing ELISA and in situ hybridization with IL-6 cDNA probe labelled by digoxingenin respectively. Results In 36/42 (85.7%) of active LN patients, uninary IL-6 content was higher than or equal to 5 pg/mg ,cr, while no one case did in normal controls. Uninary IL-6 content was highly correlated with the uninary ft-m and NAG excretion respectively. In normal tubules and interstitium, IL-6 mRNA was undetectable. For all sections from 15 active cases, IL-6 mRNA was found in tubular and interstitial cells, and the distribution correlated with the extent of tubulointerstitial lesions. Conclusion IL-6 is probably involved in tubulointerstitial lesions of patients with active lupus nephritis.
ABSTRACT
Objective To investigate the effects of IFN-? and IL-4 on MIP- ? production of peripheral blood mononuclear cells (PBMC) in lupus nephritis (LN) Methods MIP-1? expression in supematants was determined by Elisa and MLP-l? mRNA in PBMC was detected by RT-PCR. Results (1) IFN-? increased MIP-1? prouduction of PBMC in LN. (2) IL-4 inhibited MIP-1? production of PBMC in LN. (3) No significant effect on PBMC in controls was found by either IFN-? or IL-4. Conclusion IFN-? enhances MLP- 1? production of PBMC in LN wherease LL-4 inhibits it.
ABSTRACT
Objective To explore derivation of renocortieal prostaglandins in C-BSA nephritis. Methods The interfering effect of goat-antiplatelet IgG serum(APS)on this model was studied. Results The levels of renocortical PGE_2、6-keto-PGF_(1?) (PGI_2)、TXA_2 were all reduced in experimental group. The pathological damage of experimental group were less severe in comparison with the model group. Proteinuria of the experimental group was alleviated. Conclusion Platelet may play a pathogenetic role of C-BSA nephritis and renocortical prostaglandins partly derive from platelets.
ABSTRACT
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.
ABSTRACT
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
ABSTRACT
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
ABSTRACT
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.
ABSTRACT
AIM: To observe the expression of fractalkine, and its receptor, CX3CR1, in renal tissues of patients with diffuse proliferative lupus glomerulonephritis (WHO class IV), minimal glomerular abnormalities, and normal kidney. Meanwhile, the correlation among the expression of fractalkine, CX3CR1 and CD68-positive macrophages was investigated, and the role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was discussed. METHODS: The expressions of fractalkine, CX3CR1 and CD68 were detected immunohistochemically in kidney tissue sections obtained from twenty-one patients with WHO class IV lupus nephritis, eighteen cases with minimal glomerular abnormalities, and eight normal kidneys which were no abnormality under light microscope. RESULTS: (1) Fractalkine was generally indistinguishable in tissue sections from normal kidney and minimal glomerular abnormalities. CX3CR1-positive cells and CD68-positive macrophages were sparsely detected in the glomeruli and in the cortical interstitium. (2) There were considerable CX3CR1-positive cells and macrophages in both the glomeruli and the interstitium in sections from class IV lupus nephritis. The number of CX3CR1-positive cells significantly correlated with the number of macrophages in the glomeruli and in the interstitium respectively (r=0.956, P