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

NF-κ B是一种多效性转录因子，其激活许多细胞因子、生长因子和粘附分子，这些细胞因子、生长因子和粘附分子与肾小球肾炎中的细胞对损伤的反应（例如系膜细胞（MC）增殖或炎性细胞浸润）有关。本研究采用糖皮质激素、抗氧化剂（PDTC）和含有NF-κ B结合位点的寡核苷酸（decoy DNA）抑制NF-κ B的活性，观察其对体外培养的肾小球系膜细胞增殖的抑制作用和对大鼠抗Thy 1.1肾炎模型的治疗作用。首先，在体外评估这三种试剂对MC增殖的影响。从用这些试剂处理的细胞制备核提取物。电泳迁移率变动分析（EMSA），以确认对NF-κ B DNA结合活性的影响。其次，对这三种试剂进行了体内实验。用抗Thy1.1a单克隆抗体建立大鼠抗Thy1.1模型 关于我们 抗体（oX-7）。在第2天，在第8天进行肾活检。每个肾小球横截面的总细胞数和增殖核细胞抗原（PCNA）阳性细胞的数量进行了测定。提取肾小球mRNA，采用逆转录聚合酶链反应（RT-PCR）方法检测NF-κ B调控的IL-1、TNF-α、MCP-1和ICAM-1的mRNA表达。NF-κ B decoy在体外以剂量依赖的方式抑制MC的生长。10 mM诱饵抑制MC生长75%。EMSA显示，这种抑制作用是通过降低NF-κ B结合活性介导的。糖皮质激素，PDTC和NF-κ B诱饵抑制MC增殖在Thy1.1肾小球肾炎模型。肾小球细胞数减少25%。实验组IL-1、TNF-α、MCP-1、ICAM-1 mRNA表达降低。因此，系膜细胞增殖的抑制可能是由于这些细胞因子的表达至少部分受到抑制。这些结果提示靶向NF-κ B功能的试剂作为肾小球疾病的新型治疗剂的可行性。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