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

NF-kB 在肾小球肾炎发病机制中的作用及其调节的治疗策略。

基本信息

批准号：
09671167
负责人：
KASHIHARA Naoki
金额：
$ 1.98万
依托单位：
KAWASAKI MEDICAL SCHOOL (1998)Okayama University (1997)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671167/
关键词：
NF-kB Glomerulonephritis Glucocorticoid 転写因子ケモカインサイトカイン

项目摘要

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

NF-kB 是一种多效转录因子，可激活多种细胞因子、生长因子和粘附分子，这些细胞因子与肾小球肾炎中的细胞对损伤的反应有关，例如系膜细胞 (MC) 增殖或炎症细胞浸润。使用糖皮质激素、抗氧化剂 (PDTC) 和包含共有序列的寡核苷酸 (ODN) 检测 NF-kB 活性NF-kB结合位点（诱饵DNA）并评估体外对系膜细胞增殖的抑制作用以及在大鼠抗Thy1.1肾炎模型中的治疗作用首先，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中制备核提取物，进行电泳迁移率变化测定（EMSA）以确认对 NF-kB DNA 结合活性的影响。大鼠抗Thy1.1模型是通过注射单克隆抗Thy1.1抗体（oX-7）诱导的，第8天进行肾活检。测定每个肾小球横截面的增殖核细胞抗原（PCNA）阳性细胞数，提取肾小球mRNA并检测IL-1、TNF-a、MCP-1和MCP-1的mRNA表达。通过逆转录酶聚合酶链式反应(RT-PCR)方法测量了ICAM-1的基因表达受NF-kB调节。10 mM的诱饵以剂量依赖性方式抑制MC生长。 EMSA 表明，这种抑制作用是由糖皮质激素、PDTC 和 NF-kB 诱饵的抑制作用介导的。 Thy1.1肾小球肾炎模型中的MC增殖实验组中肾小球细胞的数量减少了25%，从而抑制了IL-1、TNF-a、MCP-1和ICAM-1的表达。系膜细胞增殖可能至少部分是由这些细胞因子的表达抑制引起的。这些结果表明靶向 NF-kB 功能的试剂作为新型治疗剂的可行性。肾小球疾病较少。