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

NF-kB是一种多营养转录因子，可激活多种细胞因子、生长因子和粘附分子，参与细胞对损伤的反应，如肾小球肾炎中系膜细胞（MC）的增殖或炎症细胞的浸润。我们尝试用糖皮质激素、抗氧化剂（PDTC）和含有NF-kB共同结合位点（诱饵DNA）的寡核苷酸（ODN）来抑制NF-kB活性，并在体外评估了对肾小球系膜细胞增殖的抑制作用和对大鼠抗thy1.1肾炎模型的治疗作用。首先，在体外评估这三种试剂对mcc增殖的影响。用这些试剂处理的细胞制备核提取物。电泳迁移位移试验（EMSA）证实了对NF-kB DNA结合活性的影响。其次，这三种试剂在体内使用。通过注射抗thy1.1 a…More单克隆抗体（oX-7）建立大鼠抗thy1.1模型。第2天，第8天进行肾活检。测定肾小球横切面细胞总数和增殖核细胞抗原（PCNA）阳性细胞数。提取肾小球mRNA，采用逆转录酶聚合酶链反应（RT-PCR）法检测NF-kB调控基因表达的IL-1、TNF-a、MCP-1和ICAM-1 mRNA的表达。NF-kB诱骗剂对体外mcc生长有剂量依赖性。10 mM诱捕剂可抑制mc75 %的生长。EMSA显示，这种抑制作用是由NF-kB结合活性降低介导的。糖皮质激素、PDTC和NF-kB诱饵抑制了Thy1.1型肾小球肾炎模型中MC的增殖。肾小球细胞数量减少25%。实验组IL-1、TNF-a、MCP-1、ICAM-1 mRNA表达降低。因此，系膜细胞增殖的抑制可能至少部分是由于这些细胞因子的表达受到抑制。这些结果表明靶向NF-kB功能的试剂作为一种新的肾小球疾病治疗剂是可行的。少

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