SMAD2调控ERK通路干预M2巨噬细胞活化在糖尿病肾病小鼠肾脏纤维化中的作用及机制

项目名称： SMAD2调控ERK通路干预M2巨噬细胞活化在糖尿病肾病小鼠肾脏纤维化中的作用及机制

项目编号： No.81500552

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 苏宁

作者单位： 中山大学

项目金额： 18万元

中文摘要： 巨噬细胞与糖尿病肾病(DN)发病密切相关，我们前期研究发现:DN小鼠肾脏M2型巨噬细胞渗出增多，并参与了肾脏纤维化进程，且巨噬细胞向M2分化过程中伴有ERK1/2及SMAD2活化，但阻断ERK仅可部分阻断巨噬细胞向M2分化，且阻断ERK对SMAD2活化无影响。SMAD2是否为DN肾脏巨噬细胞向M2分化的关键通路，且ERK是否受控于SMAD2活化目前尚不清楚。本研究中，我们将构建SMAD2、 SMAD7正义及反义质粒：①通过超声微泡技术转染DN小鼠，观察其肾脏巨噬细胞向M2分化是否受SMAD2调控，且是否伴随M2分化的减弱存在肾脏纤维化程度减低。②体外转染RAW264.7巨噬细胞株，并以高糖及TGF-β1诱导其向M2分化，观察ERK1/2活化是否受控于SMAD2的活化。通过该研究提出DN新的发病机制，为临床通过干预SMAD2及ERK1/2活化治疗DN提供新的靶点。

中文关键词： 糖尿病肾病；巨噬细胞；信号通路；SMAD2；ERK

英文摘要： Macrophages are associated with the process of diabetic nephropathy.M2 macrophages are involved in the kidney fibrosis of diabetic mice in our previous research. ERK and SMAD2 are all activated in M2 macrophages. But the process of M2 macrophage can not be fully blocked by the use of ERK inhibitor, and the activation of SMAD2 can not also be inhibited with it. The effect of SMAD2 pathway in the process of M2 macrophage of diabetic mice, and the relationship between the ERK and SMAD2 in M2 macrophage are not clear. In this research, the plasmid of SMAD7 ,shSMAD7,SMAD2 and shSMAD2 would be constructed and transfected to the diabetic mice with the use of ultrasound microbubble, then the renal M2 macrophages and the renal fibrosis would be measured ,which will confirm the effect of SMAD2 in the process of M2 macrophages.Furthermore,RAW264.7 macrophages would be transfected with these plasmids in vitro, then the macrophages will be induced to M2 phenotype with the high ambient glucose and TGF-β1,and the activation of ERK and the ratio of M2 phenotype in these macrophages will be investigated, which will confirm the relationship between ERK and SMAD2.From this research, a new pathogenesis of diabetic nephropathy will be found ,as a result, a new target to treat the diabetic nephropathy will also be discovered.

英文关键词： diabetic nephropathy;macrophage;signal pathway;SMAD2;ERK