[Interleukin 1 beta gene expression by mouse glomerular mesangial cells]

Abstract

Interleukin 1 beta (IL-1 beta) plasmid DNA was isolated and purified with alkaline lysis method and equilibrium centrifugation in cesium chloride. IL-1 beta plasmid RNA was cut by restriction endonucleases EcoRI and XbaI, and after that it was 1 loaded in 0.75% low-melting-temperature agarose and underwent electrophoresis. 1.2 Kb DNA fragments were extracted from gel and for further purification it was used as IL-1 beta cDNA probe. RNA from mouse glomerular mesangial cells, tubular epithelial cells and P388D, cells was cut with EcoRI separately and then hybridized in dot blots with alpha 32P labeled IL-1 cDNA probe. The dot blot autoradiograph showed expression of the IL-1 beta gene in mesangial cell and P388D1 cell but showed no expression in epithelium. These results not only identified the specificity of IL-1 beta probe but also suggested that the local production of IL-1 beta may be important in the pathogenesis of glomerulonephritis.