转录的超保守区域uc.153作为ceRNA调控慢性炎性疼痛的作用及机制研究

Aberrant gene expression is an important cause of chronic pain, however, the gene expression regulatory mechanism associated with chronic pain is still unclear. Transcribed ultraconserved region (T-UCR), a newly discovered way of gene expression regulation, is involved in a variety of pathological and physiological processes. Our previous experiments found that complete Freund's adjuvant (CFA) induced-chronic peripheral inflammation caused a significant increase in spinal uc.153 expression, knockdown of uc.153 expression reduced chronic inflammatory pain behaviors; by further experiments, we found that uc.153 regulate inflammatory pain may be through interacting with miR-327 and influencing the expression of its target gene tet methylcytosine dioxygenase 3 (TET3). Therefore, according to the existing literature and our previous experiments, we hypothesized that uc.153 was increased in the spinal after chronic peripheral inflammation, it regulates chronic inflammatory pain by functioning as a competitive endogenous RNA (ceRNA) to compete with TET3 for miR-327 binding. By using a CFA-induced chronic inflammatory pain model, this project intends to confirm the above hypothesis at the whole body, cell and molecular levels. This work will explore the mechanism of chronic pain from the new perspective of T-UCR, and provide a novel thought for preventing and treating chronic pain.

基因表达异常是慢性痛产生的重要原因，但与慢性痛相关的基因表达调控机制仍不清楚。转录的超保守区域（T-UCR）是近年来发现的一类新的基因表达调控方式，参与多种病理生理过程。我们前期预实验发现，完全弗氏佐剂（CFA）诱导的外周慢性炎症能显著上调脊髓uc.153的表达水平，抑制uc.153表达可以减轻慢性炎性痛行为；进一步研究，我们发现uc.153可能通过与miR-327相互作用，影响其靶基因甲基胞嘧啶双加氧酶3（TET3）表达参与慢性炎性痛的调控。因此，根据已有文献及课题组预实验，我们提出假设：外周慢性炎症能够引起脊髓水平uc.153表达增加，其作为竞争性内源RNA（ceRNA）与TET3竞争性的结合miR-327参与调控慢性炎性痛。本项目拟以CFA诱导的慢性炎性痛为模型，从整体、细胞和分子水平对以上假设进行验证。本课题将从T-UCR这个新视点探讨慢性痛的发生机制，为慢性痛的防治提供新的思路。

转录的超保守区是一类新的长非编码RNA，在人、大鼠和小鼠中是完全保守的。转录的超保守区域与多种生物过程有关，但是，目前对其在疼痛调控中的作用知之甚少。在本项目的资助下，通过运用电生理学、动物行为学、分子生物学等多种手段和方法相结合的措施，在整体动物水平、细胞水平和分子水平对转录的超保守区uc.153进行了系统地研究，结果发现：1）CCI诱导的神经病理性疼痛能够引起脊髓uc.153的表达增加；2）调控脊髓uc.153的表达能够调节小鼠的疼痛行为；3）miR-182-5p，而非原先发现的miR-237，是uc.153的下游靶点；4）uc.153通过调节miR-182-5p前体（pre-miR-182-5p）的成熟过程调控神经病理性疼痛。我们的发现揭示了uc.153在疼痛调节中的重要作用，并为神经性疼痛治疗提供了一个新的药物靶点。