中文摘要
焦虑症为常见的精神障碍之一，对其发病机制和防治的研究一直是该领域的研究热点。胞质多聚腺苷酸化元件结合蛋白（CPEBs）是一组介导mRNA胞质多聚腺苷酸化和翻译的RNA结合蛋白，参与细胞发育、分化与衰老、以及中枢突触可塑性调节等过程。我们前期研究发现CPEB1在与焦虑相关的神经核团如杏仁核和前扣带回高表达，采用杏仁核注射慢病毒CPEB1-shRNA可显著降低小鼠束缚应激引起的焦虑样行为，提示CPEB1及其功能紊乱在焦虑症的发病中具有重要作用。本项目首次提出在慢性应激条件下，杏仁核CPEB1调控突触内蛋白过度翻译可能是引起兴奋性/抑制性传递平衡紊乱的重要分子机制，并参与了焦虑症的病理发生过程。本项目结合电生理、分子生物学和行为学方法，探讨CPEB1在兴奋性和抑制性突触内对蛋白翻译的调控，为深入了解焦虑症的病理机制和发掘药物作用新靶点，提供新的思路和理论依据。
英文摘要
Anxiety is one of the most common mental disorders. The research on the pathogenesis and prevention of anxiety is a hot topic in this field. Cytoplasmic polyadenylation element-binding proteins (CPEBs), a group of mRNA binding proteins which mediate cytoplasmic polyadenylation and translation of the RNA binding protein, involve in cell growth, differentiation and aging, as well as the central synaptic plasticity modulation. Our previous study found that CPEB1 was highly expressed in the anxiety associated nucleus such as the amygdala and anterior cingulate cortex. Amygdala microinjection of CPEB1-shRNA could significantly reduce the anxiety-like behavior induced by restraint stress in mice. These results suggest that CPEB1 and its dysfunction play an important role in the pathogenesis of anxiety disorder. The present project proposes that the amygdala CPEB1 may be an important molecular in regulating synaptic protein overexpression in the chronic stress condition and the balance between excitatory/inhibitory transmission, and involve in the pathogenesis of anxiety disorder. Multiple methods including electrophysiology, molecular biology and behavioral tests will be used to elucidate the roles of CPEB1 in the regulation of excitatory and inhibitory synapse protein translation. It will be helpful to understand the mechanisms underlying the pathogenesis of anxiety disorder and to provide a theoretical basis for exploring new anxiolytic drug targets.
结题摘要
焦虑症为常见的精神障碍之一，对其发病机制和防治的研究一直是该领域的研究热点。胞质多聚腺苷酸化元件结合蛋白（CPEBs）是一组介导mRNA胞质多聚腺苷酸化和翻译的RNA结合蛋白，参与细胞发育、分化与衰老、以及中枢突触可塑性调节等过程。我们前期研究发现CPEB1在与焦虑相关的神经核团如杏仁核和前扣带回高表达，采用杏仁核注射慢病毒CPEB1-shRNA可显著降低小鼠束缚应激引起的焦虑样行为，提示CPEB1及其功能紊乱在焦虑症的发病中具有重要作用。本项目首先建立了束缚和慢性痛引起小鼠焦虑的动物模型，采用行为学、Western blot和CPEB1-shRNA转染技术等方法检测了CPEB1参与杏仁核网络功能的调节和在焦虑中的作用。成年雄性小鼠，连续束缚两日后，进行高架十字和旷场行为学的评估，发现小鼠在开臂和中央区活动时间明显减少，表现出明显的焦虑样行为，取小鼠杏仁核并制备样本做Western blot，结果显示束缚应激组小鼠杏仁核中CPEB1受体表达明显增加。此外，我们还建立了内脏痛引起的小鼠焦虑动物模型，小鼠表现出明显的焦虑样行为，并持续至少4周时间，杏仁核中的CPEB1持续高表达。此外，内脏痛模型组小鼠前皮层扣带回和岛叶中CPEB1表达也明显增加，但是海马、丘脑和躯体皮层中CPEB1的表达与对照组相比没有变化。为了证实CPEB1参与焦虑调控中的特异性，我们建立了急性痛和恐惧记忆动物模型，发现杏仁核中CPEB1的表达没有变化。由于焦虑发生与突触前谷氨酸释放增加密切相关，因此我们体外培养了神经元细胞，谷氨酸孵育1h可引起CPEB1表达明显增加，结果与体内相似。以上结果提示：CPEB1参与了焦虑的调控。采用CPEB1-shRNA慢病毒转染技术，发现CPEB1-shRNA慢病毒在1:20大约减少CPEB1表达50%左右，将慢病毒脑定位微量注射至小鼠杏仁核部位能够明显缓解束缚和慢性痛引起的焦虑样行为。