基于miRNA调控的“蒙医三根平衡针法”调节抑郁大鼠BDNF-tPA-P11信号通路的分子机制研究

Depression is one of the most common feature of mental disorders, lack of high reliability diagnostic indicator and specific treatment up until now. Mongolian medicine consider depression as a cause of overactive of Heyi and Badagan, blood phase stroke, He Yi, dysfunction of blood metabolism, blocking the white pulse (belongs to the nervous system).Mongolian Sangen balance treatment on depression has the characteristics of less blood site and its efficiency. It has been reported to mediate the BDNF signaling the rat hippocampus. Our previous study showed that BDNF is one of the target of micro RNA, while the miRNA plays an important role in the development and treatment of depression. This project will use Mongolian sangen balance method to intervene “ Heyi Acupuncture point”, “Badagan point” and “heart point” in depression rats. Combined with behavioral analysis, miRNA microarray to understand the depression like behavior in CUMS rat and the effect of Mongolian Sangen balance treatment on expression of miRNA microarray. Molecular biology analysis will be conducted to identify the changes of molecules involved in the BDNF-tPA-P11 signaling pathway. This will allow us to explore the mechanism of mediating BDNF-tPA-P11 signaling pathway by miRNA target to BDNF, which will provide an effective idea and method to inheritance and innovation of Mongolian specific therapy.

抑郁症是最常见的精神障碍之一，目前尚缺乏高信度的诊断指标和特效性的治疗手段。蒙医学认为，抑郁症是体内赫依偏盛与巴达干、血相搏，赫依、血运行不畅，阻塞白脉（属神经系统）之传导所致。蒙医三根平衡针法治疗抑郁症取穴少、疗效显著的特点，对抑郁大鼠海马BDNF有调控作用。我们在前期研究中发现，BDNF是miRNA-16的靶标之一，miRNA-16在抑郁发生及抗抑郁治疗中发挥重要作用。本课题以蒙医三根平衡针法干预抑郁大鼠赫依穴、巴达干穴、心穴为切入点，通过行为学评价、miRNAs芯片技术，明确CUMS大鼠出现抑郁样行为以及蒙医三根平衡针对抑郁大鼠miRNAs表达谱的影响；采用分子生物学技术观察抑郁大鼠miRNA-16与BDNF-tPA-P11通路的相关分子表达变化，探讨miRNA-16靶向BDNF进而调控BDNF-tPA-P11通路的作用机制，为蒙医特色疗法的传承与创新提供行之有效的思路和方法。

抑郁症是常见的精神障碍之一，蒙医三根平衡针法治疗抑郁症疗效确切，尤其改善抑郁症躯体化症状方面疗效更为显著，但作用机制尚不明确。本课题以蒙医三根平衡针法干预抑郁模型大鼠赫依穴、巴达干穴、心穴为切入点，通过行为学评价、miRNAs芯片技术、分子生物学技术观察抑郁模型大鼠海马和中缝核组织miRNAs表达谱及miRNA-16与BDNF-tPA-P11通路的相关分子表达变化。结果显示，在正常组与模型组间得到25条差异表达miRNA，这些miRNA的靶基因与细胞内信号转导、蛋白质磷酸化、大脑发育、心脏发育等生物学进程有关,与轴突导向、癌症通路、cGMP-PKG信号通路、神经营养因子信号通路的关系最强。而在正常组、模型组、药物组、针刺组之间得到65条差异表达miRNA。差异表达miRNA与Nlgn1、Clock、Per3、Plin2等基因的关系最强；这些基因与神经发育、生物节律过程、光刺激反应等生物学功能有关；与轴突导向、昼夜节律、多巴胺能突触、神经营养因子通路的作用关系最强。与正常组比较，模型组大鼠p11和tPA阳性表达减少；与模型组比较，药物组、蒙医针组大鼠p11和tPA阳性表达增加；模型大鼠海马和中缝核区miRNA-16表达增加，而BDNF表达减少。与模型组比较，药物组和蒙医针组miRNA-16表达减少，BDNF表达增加。提示，蒙医三根平衡针法能够改变大鼠海马miRNAs表达谱，抑制miRNA-16的表达。miRNA-16能靶向调控BDNF表达，BDNF、tPA、P11在表达和功能上有着相互作用关系，并在抑郁发作和抗抑郁治疗中起着至关重要的作用。蒙医三根平衡针法可能通过抑制miRNA-16表达上调其BDNF表达，进而调节BDNF-tPA-P11信号通路起到抗抑郁效应。该研究揭示了蒙医三根平衡针法治疗抑郁症潜在靶点和作用机制，为蒙医特色疗法的传承与创新具有重要的提示作用。