The role of NF-kB in the pathogenesis of glomerulonephritis and therapeutic strategy through its regulation.

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NF-kB is a pleiotrophic transcription factor, which activate a number of cytokines, growth factors and adhesion molecules, which are implicated in cellular response to injury , such as mesangial cell (MC) proliferation or infiltration of inflammatory cells, in glomerulonephritis. We attempted to inhibit NF-kB activity by using glucocorticoid, anti-oxidant (PDTC), and oligonucleotide (ODN) which contains the consensus NF-kB binding site (decoy DNA) and evaluated inhibitory effect on mesangial cell proliferation in vitro and therapeutic effects in the rat anti-Thy1.1 nephritis model as well. First, the effect of these three reagents on MC proliferation was assessed in vitro. The nuclear extracts were prepared from the cells treated with these reagents. Electrophoretic mobility-shift assay (EMSA) was performed to confirm the effect on NF-kB DNA binding activity. Secondly, these three reagents were used in vivo. The rat anti-Thy1.1 model was induced by injection of monoclonal anti-Thy1.1 a … More ntibody (oX-7). At day 2, Renal biopsy was performed at day 8. The total cell number and the number of proliferating nuclear cell antigen (PCNA)-positive cells per glomerular cross section were determined. Glomerular mRNA was extracted and the mRNA expressions of IL-1, TNF-a, MCP-1 and ICAM-1 of which gene expression was regulated by NF-kB, were measured by reverse transcriptase polymerase chain reaction (RT-PCR) method. NF-kB decoy inhibited the MC growth in a dose dependent manner in vitro. Ten mM of decoy inhibited the MC growth by 75%. EMSA revealed that this inhibitory effect was mediated by decreased NF-kB binding activity. Glucocorticoid, PDTC and NF-kB decoy suppressed MC proliferation in the Thy1.1 glomerulonephritis model. The number of glomerular cell decreased by 25%. Decreased mRNA expressions of IL-1, TNF-a, MCP-1 and ICAM-1 were recognized in the experimental group. Thus, inhibition of mesangial cell proliferation was possibly resulted from suppressed expression of these cytokines at least in part. These results suggest the feasibility of the reagents targeting NF-kB funtion as a novel therapeutic agent for glomerular diseases. Less