胰岛β细胞miRNA信号放大检测新方法研究

胰岛β细胞功能受损是糖尿病发生的重要病理生理机制。微小RNA(Micro RNA，miRNA)与胰岛β细胞的发育和功能损伤有关。本项目拟以糖尿病的预警及早期治疗为目标，设计构建新型核酸传感平台，有效分离分子识别单元与报告单元，提出胰岛β细胞miRNA-124a及miRNA-9的放大检测新方法，并通过对miRNA-124a及miRNA-9表达水平的实时监测，分析其与β细胞功能损伤及糖尿病发病机制的相关性。主要研究内容包括：1)胰岛β细胞离体培养及miRNA的提取；2)构建基于发夹分子开关及三链分子开关的核酸探针识别单元；3)结合核酸工具酶及功能化纳米材料实现信号转换及多重放大。本项目的执行将提出胰岛β细胞miRNA放大检测的新型核酸传感体系，促进胰岛β细胞miRNA检测方法的发展，为糖尿病早期诊断和预防提供参考。

Progressive loss of the function of β-cell is believed to be a causative factor in diabetes. The relationship between the expression level of miRNA and β-cell development as well as its dysfunctions have been discussed in recent researches. In this project, the prevention and treatment of early diabetes is aimed as a breakthrough, new molecular recognition and signal amplification platform combing with functional nanomaterials will be constructed to achieve ultrasensitive and real-time monitoring of miRNA-124a and miRNA-9 expression in β-cell. This project focuses on the following three aspects: 1)Culture β-cell in vitro and extract miRNA. 2)Construct molecular recognition platform based on hairpin-structured DNA molecular switch and triplex-structured DNA molecular switch. 3)Construct signal transduction and multiple amplification strategies combing with functional nanomaterials and nuclease. From this project, new design for β-cell miRNA detection will be proposed, which will promote the development of β-cell miRNA sensing, provide new means for early diagnosis and prevention of diabetes.