JNK1和JNK2在腹膜透析相关性腹膜纤维化中的作用及机制研究

JNK在细胞分化、凋亡及基因表达中起重要作用。我们已发现JNK介导了腹膜间皮细胞EMT，且JNK1与Smad3存在交互作用。但JNK1和JNK2在腹膜纤维化中的各自作用及机制尚不清楚。本项目研究如下：在JNK1或JNK2基因敲除小鼠，采用5/6肾切除建造尿毒症模型，腹腔注射高糖腹透液，观察JNK1/2在腹膜纤维化中的作用及其对Smad通路的影响；分离野生型、JNK1或JNK2基因敲除小鼠原代腹膜间皮细胞，观察TGF-β1刺激下，AP-1组成分子c-Jun和c-fos的表达，以及EMT指标的变化；进一步在小鼠原代腹膜间皮细胞，转染c-Jun或（和）c-fos siRNA，或使用AP-1抑制剂T-5224，观察其对Smad3与AP-1分子共定位、Smad3与Snail启动子序列结合及EMT的影响。本项目旨在明确不同亚型JNK在腹膜纤维化中作用及分子机制，为防治腹透相关性腹膜纤维化提供新的靶点。

Many studies suggested that the JNK signal pathway plays a pivotal role in cell differentiation, apoptosis, and a variety of genes expression. Our previous studies have demonstrated that JNK involves in TGF-β1 induced rat peritoneal mesothelial cells EMT, and there is a mutual interaction between the TGF-β-related JNK1 and Smad3 pathways. However, the molecular mechanism and the individual role of JNK1 and JNK2 in peritoneal fibrosis in vivo remains poorly understood. In the present study, we will examine the exact role of JNK1 and JNK2 in peritoneal fibrosis in JNK1/2 gene knock-out mice which were intraperitoneally injected with peritoneal dialysis fluid (PDF) containing 4.25% glucose after subtotal nephrectomy. In addition, we will examine the impact of JNK1/2 knock-out on the expression of c-Jun and c-fos after TGF-β1 stimulation in primary JNK1/2 mice peritoneal mesothelial cell. Then, we will examine the impact of the c-Jun and (or) c-fos siRNA or AP-1 inhibitor T-5224 on the binding of Smad3 to the snail promoter in primary mice peritoneal mesothelial cell, and the impact on EMT. This study aims to investigate the distinct role and molecular mechanism of JNK1 and JNK2 in dialysis-related peritoneal fibrosis and provide a novel therapeutic agent for peritoneal fibrosis.