Nrf2-miR-233-NR2B通路抑制硝酸甘油诱导痛觉过敏的机制研究

项目名称： Nrf2-miR-233-NR2B通路抑制硝酸甘油诱导痛觉过敏的机制研究

项目编号： No.81500965

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： 邸伟

作者单位： 西安交通大学

项目金额： 17.5万元

中文摘要： 痛觉过敏可导致偏头痛慢性转化。而我们的前期研究发现Nrf2激活剂可抑制硝酸甘油诱导的大鼠痛觉过敏，但目前Nrf2调节中枢敏化的机理尚待明确。NR2B表达增加介导中枢敏化已得到证实，而NR2B 基因的3’-UTR是miRNA-223靶点之一，最近研究发现Nrf2可上调miRNA-223的表达。据此我们推测Nrf2- miRNA-223-NR2B通路可能参与NTG诱导的痛觉过敏。本项目应用RT-PCR、免疫印迹及全细胞膜片钳等阐明NR2B在NTG诱导的中枢敏化中的作用及Nrf2可能通过NR2B参与抑制痛觉过敏；双荧光素酶报告基因、染色质免疫共沉淀等确立Nrf2、miRNA-223及NR2B的关系；进一步通过转染过表达Nrf2和/或阻断miRNA-223，观察miRNA-223、NR2B-mRNA及NR2B的变化。研究结果有望揭示Nrf2抑制痛觉过敏的分子机制并为其成为新的治疗靶点提供科学依据。

中文关键词： 核因子E2相关因子-2；微小RNA；N-甲基-D-天门冬酸受体；中枢敏化

英文摘要： Hyperalgesia may lead to migraine chronic transformation which seriously impact on the quality of life in patients. Our previous study found that Nrf2 activator attenuated hyperalgesia symptom induced by nitroglycerin, but the mechanism of Nrf2 regulating central sensitization need to identify. NR2B mediate central sensitization has been confirmed, while the 3'-UTR region of NR2B gene is one of the miRNA-223 targets. Moreover, a recent study found that Nrf2 could increase miRNA-223 expression. Accordingly, we speculate that Nrf2- miRNA-223-NR2B pathway is involved in NTG-induced hyperalgesia. In the present study, RT-PCR, western blot and whole –cell patch clamp are used to clarify the role of NR2B in the NTG-induced central sensitization and that Nrf2 are involved in inhibiting hyperalgesia through NR2B. Dual luciferase reporter gene system, chromatin immunoprecipitation are used to establish the relation between Nrf2, miRNA-223 and NR2B. Further, overexpressing Nrf2 / or blocking miRNA-223 by ransfection to observe changes in miRNA-223, NR2B-mRNA, and NR2B. Collectively, successful completion of this project might reveal the molecular mechanism of Nrf2 inhibiting hyperalgesia induced by NTG, and provide a scientific basis for a new target for the treatment of the hyperalgesia.

英文关键词： Nrf2; miRNA;NMDAR;central sensitization