多靶点microRNA干预技术调控GLUT4信号通路对糖尿病的治疗作用

Diabetes mellitus is a serious life threatening disease which is an ominous threat to public health. Decreased insulin sensitivity and barrier to tissue glucose usage are important features of type 2 diabetes mellitus. To search for early warning molecules indicating diabetes mellitus and elucidate the pathogenesis are the urgent problems to be resolved. MicroRNA is not only regulating the pathological and physiological functions of the body, but also becoming the early warning indicators of cancer and many other diseases. Whether miRNA can play a regulatory role in early state of diabetes or it can be early warning factor of DM are not yet clear. In preliminary studies, we filtered out a group of pathogenic potential of miRNAs which can target to signal pathway proteins related to glucose transporter protein(GLUT4). This study is going to explore the variety of these miRNAs in early stage of diabetic patients and diabetic animal models, to make sure whether these miRNAs can be early warning factors and to clarify molecular signaling pathways. This study will also apply multi-target miRNA antisense nucleotide inhibitors (MTg-AMO) technology we firstly established to investigate its effect on tissue glucose utilization, to expand our understanding of therapeutic intervention role of miRNA and to provide a new therapeutic strategy for diabetes mellitus .

糖尿病严重威胁人类健康，胰岛素抵抗是2型糖尿病发生、发展的一个重要特征，寻找2型糖尿病发病早期预警分子并阐明其发病机制是目前临床亟需解决的热点问题。microRNA不仅调节机体的病理、生理功能，同时也已成为肿瘤等疾病的早期预警指标。糖尿病早期miRNA是否发挥调节作用，其能否成为糖尿病发生的早期预警因子尚不清楚。在前期研究中我们运用生物信息学等手段筛选出一组具有致病潜能的miRNAs，这些miRNAs可分别靶向与葡萄糖转运蛋白GLUT4相关的信号通路蛋白。本项目拟在2型糖尿病早期患者和动物模型上验证这组miRNAs的变化，探讨这组miRNAs能否成为糖尿病早期预警因子，并阐明其分子信号通路。本项目还将首次应用我们建立的多靶点miRNA反义核苷酸抑制剂（MTg-AMO）技术，探讨其对于组织葡萄糖摄取利用能力的改善作用，拓展我们对于miRNA干预治疗的认识，为糖尿病治疗提供新的策略。

本项目按计划书顺利完成，我们首次发现miRNA-106b, miR-27a和miR-30d可分别调控GLUT4、p38 MAPK和PIK3R2的表达，影响胰岛素抵抗。2型糖尿病大鼠骨骼肌和胰岛素抵抗的细胞模型中，GLUT4、p38 MAPK和PIK3R2的表达增加，而miRNA-106b, miR-27a和miR-30d的表达减少。应用luciferase报告基因技术检测发现miRNA-106b, miR-27a和miR-30d可分别与GLUT4、p38 MAPK和PIK3R2的3’UTR结合，显著抑制荧光素酶活性。分别抑制miRNA-106b, miR-27a和miR-30d可明显增加葡萄糖的摄取。建立多靶点miRNA 反义核苷酸抑制剂（MTg-AMO）技术，结果表明MTg-AMO可增加GLUT4、p38 MAPK和PIK3R2的表达增加，降低miRNA-106b, miR-27a和miR-30d的水平，增加葡萄糖的摄取。