基于LncRNA NFE2L2/miR-1931探讨益气养阴活血法调控糖尿病肾病内质网自噬的机制研究

前期工作发现益气养阴活血法可以显著改善糖尿病肾病蛋白尿，提高内质网自噬水平。采用LncRNA测序技术及生物信息学方法筛选出内质网自噬相关通路LncRNA NFE2L2/miR-1931。本项目拟应用db/db小鼠作为糖肾模型，给予高糖刺激小鼠肾小球系膜细胞建立高糖损伤细胞模型，体内、体外分别过表达和抑制LncRNA NFE2L2，观察下游基因及内质网自噬活性变化；其次，采用双荧光素酶报告实验、Ago2RIP方法检测LncRNA NFE2L2与miR-1931的海绵作用。体外同时抑制LncRNA NANFE2L2和miR-1931，观察系膜细胞内质网自噬活性变化。最后，在此基础上，采用益气养阴活血法干预db/db小鼠及体外高糖损伤系膜细胞，阐明益气养阴活血法对LncRNA NFE2L2/miR-1931及内质网自噬的干预效应，这将为中医药有效防治糖尿病肾病提供新的理论基础和治疗靶点。

In our previous studies, the results showed that Yiqi Yangyin Huoxue therapy can significantly improve proteinuria and endoplasmic reticulum autophagy in diabetic nephropathy. Meanwhile, we obtained the LncRNANFE2L2/miR-1931 pathway related to endoplasmic reticulum autophagy by LncRNA sequencing and bioinformatics. In this project, we intend to use db/db mice as diabetic nephropathy mice model and mice mesangial cells cultured in high glucose condition as high glucose injury cell model. We plan to over express and inhibit LncNFE2L2 in vivo and in vitro and observe the expression levels of downstream genes and endoplasmic reticulum autophagy activity. Secondly, the sponge effect of LncNFE2L2and miR-1931 was detected by double Luciferase Reporter assays and Ago2 RIP method. In addition, LncNFE2L2 and miR-1931 were inhibited simultaneously in vitro and the activity of endoplasmic reticulum autophagy was observed. Finally, on this foundation, we plan to apply Yiqi Yangyin Huoxue therapy to intervene db/db mice and mesangial cells cultured in high glucose condition to clarify the effects of Yiqi Yangyin Huoxue therapy in the regulation of LncRNANFE2L2/miR-1931 pathway and the activity of endoplasmic reticulum autophagy. It will provide new theoretical basis and therapeutic target for the prevention and treatment of diabetic nephropathy by traditional Chinese medicine.