基于自噬探讨“痰、郁”病机与癫痫-抑郁共病的相关性及柴胡加龙骨牡蛎汤的干预作用

Epilepsy-Depression Comorbidity (EDC) is secondary to epilepsy and accounts for 23.1% in epileptic patients. It is the main reason of premature death in epileptic patients. The pathogenesis of EDC can be summarized by “Phlegm, Melancholia”. Phlegm and Stasis can change into Melancholia gradually, at the same time Melancholia will in turn aggravate Phlegm and Stasis. Autophagy has been regarded as Phlegm and Stasis at the microscopic level of the cell, which affects hippocampal neural regeneration directly. Therefore, we infer that the pathogenesis of EDC in state of “Phlegm, Melancholia” is related to the inhibition of hippocampal neural regeneration induced by abnormal autophagy. In our previous study, Chaihu-Longgu-Muli Decoction (CLMD) can remarkably relieve symptoms of both epilepsy and depression in EDC patients, meanwhile decrease the apoptosis ratio of hippocampal neurons in EDC rats. In this project, losts of detection methods including macroscopic representation of Chinese Medicine, behavioral detection, immunohistochemisty, Western blot and RT-PCR will be used. This study will be carried out at the level of “overall-cell-molecular” and “shape-function”, with experiments in vivo and in vitro designed altogether. The reasons of relieving EDC symptoms will be explored wherther by Up-regulating the 5-HT1A Receptor, mediating the mTOR Pathway, affecting autophagy and then increasing hippocampal neural regeneration or not. The results will reveal the molecular mechanism of “soothing the liver and regulating qi, resolving phlegm and relieving melancholia” of CLMD and offer novel treatments for EDC with Traditional Chinese Medicinal herbs.

癫痫-抑郁共病（EDC）继发于癫痫，占癫痫患者的23.1%，是癫痫患者过早死亡的主要原因。EDC的中医病机可用“痰、郁”概括，痰瘀可转郁，郁又加重痰瘀。自噬是中医痰瘀在细胞微观层面上的体现，直接影响海马神经再生。据此我们推测：EDC的“痰、郁”病理机制与细胞自噬功能异常导致海马神经再生受到抑制密切相关。我们前期研究证实，柴胡加龙骨牡蛎汤（CLMD）可有效缓解EDC患者的癫痫和抑郁症状，降低EDC大鼠海马神经元凋亡率。本项目拟采用中医宏观表征观察、行为学检测、免疫组化、Western blot、RT-PCR等方法，从“整体-细胞-分子”三层面，“形态-功能”两角度，结合体内外实验，探索CLMD是否通过上调5-HT1A受体，介导mTOR通路，调控细胞自噬，增强海马神经再生，最终缓解EDC相关症状，从而实现其“疏肝理气、化痰解郁”作用的分子机制，为中医药防治EDC提供新的思路和方法。

癫痫-抑郁共病（EDC）的中医病机可用“痰、郁”概括，痰瘀可转郁，郁又加重痰瘀。自噬是中医痰瘀在细胞微观层面上的体现，直接影响海马神经再生。据此我们认为，EDC的“痰、郁”病理机制与细胞自噬功能异常导致海马神经再生受到抑制密切相关。本研究通过制备大鼠EDC模型，采用行为学检测、免疫荧光、电镜、Western-blotting、RT-PCR等方法，探索CLMD是否通过介导mTOR通路，调控细胞自噬，增强海马神经再生，最终缓解EDC相关症状，从而实现其“疏肝理气、化痰解郁”作用的分子机制。. 结果显示：①EDC大鼠体重、摄食量、水平运动次数、垂直运动次数均较正常大鼠明显降低，经过氟西汀或CLMD治疗后，各项指标均明显提升；②EDC大鼠海马FG标记阳性神经元数目明显低于与正常组，经过氟西汀组或CLMD治疗后海马FG标记阳性神经元数目明显上升；③CLMD能明显降低癫痫大鼠癫痫发作次数，以高剂量组效果更显著；④CLMD能明显降低癫痫大鼠海马齿状回自噬体的数量，改善自噬，特别是高剂量使用时效果更为显著；其作用机制可能与促进mTOR的表达，抑制Beclin-1和LC3的表达有关；⑤CLMD对颞叶癫痫患者有明显的治疗作用，显著增加血清SOD、NGF、BDNF水平，降低MDA，减少痫样放电。. 本研究既为EDC的中医药治疗提供实验依据和科学参考，又为癫痫的临床治疗提供了新的思路。