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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 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 find new gene function associate with active lupus nephritis (LN) through study on the difference in gene expression of peripheral blood mononuclear cells between LN patients and healthy controls by gene chip. METHODS: The CSC-GE-80 chip containing 8 000 spots of cDNAs were used to investigate the difference of the expression. Both the total RNA from peripheral blood mononuclear cells of active LN patients and healthy donors were reversely transcribed to cDNA with the incorporation of fluorescent( cy3 and cy5) labeled dCTP to prepare the hybridization probes. After hybridization, the gene chip was scanned for the fluorescent intensity. The differentially expressed genes were screened. We repeated that in three groups of LN patients and healthy controls, respectively, and only the genes that have differential expression in all three chips were considered associated with LN. RESULTS: 75 genes were identified to be differently expressed in all three groups of LN patients as compared with healthy controls, including 42 up-regulated genes and 33 down-regulated ones. CONCLUSION: The present study represents a global view of gene expression of LN and provides important clues for further study of LN related genes. And it also suggests defensin ? 1, S100A8, S100A9 may be involved in the pathogenesis of LN.
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AIM: To investigate the effects of interleukin-13(IL-13) on the inflammatory reaction of glomerular mesangial cells. METHODS: TNF-? protein in the supernatant of cultured mesangial cells was determined with ELISA. ICAM-1 expression on the mesangial cells was determined with flow cytometry. The expression of TNF-? mRNA and ICAM-1 mRNA were semiquantatively tested with RT-PCR. RESULTS: Mesangial cells did not constitutively express TNF-? mRNA and TNF-? protein. Induced by LPS(10 mg/L) for 24 h, mesangial cells expressed TNF-? mRNA and TNF-? protein at a high level. At the concentration of 1 ?g/L and 10?g/L, IL-13 markedly inhibited LPS-induced TNF-? mRNA and protein expression in mesangial cells. At the concentration of 100?g/L, IL-13 completely inhibited LPS-induced TNF-? mRNA and protein expression in mesangial cells. ICAM-1 constitutively expressed on the surface of mesangial cells at very low level. Induced by TNF-?(100?g/L), mesangial cells expressed ICAM-1 mRNA and ICAM-1 cell surface molecule at high level. Costimulated mesangial cells with IL-13(10?g/L) and TNF-?(100?g/L), IL-13 inhibited TNF-?-induced ICAM-1 mRNA and cell surface molecule expression at every time course. CONCLUSION: IL-13 not only inhibits LPS-induced TNF-? expression but also inhibits TNF-?-induced ICAM-1 expression in mesangial cells. These data suggest that IL-13 inhibits the inflammatory reaction in mesangial cells.
