解郁安神颗粒治疗中风后抑郁的经典与非经典神经网络机制

Depression is a common occurrence after stroke and is associated with excess disability, cognitive impairment, and mortality. Post-stroke depression (PSD) prevalence rates range from 30% to 50%. In Traditional Chinese Medicine, depression syndrome is the major menifest in PSD. The pathogenesis of depression is a stagnation of liver Qi, and relieving Qi stagnancy is the main therapeutic principle. It is reported that resolving stagnation for tranquilization is effective method for treating depression in clinical practice. The efficacy of Jieyu Anshen pellet (JY) is dispersing stagnated liver Qi for relieving Qi stagnation and tranquilization. Our previous results demonstrated that JY ameliorate PSD in stroke model of mice and rats. However, the characteristics and mechanisms of pharmacological action of JY are still unclear. The objective of the present study is to assess the effect of JY on neurotransmitter, transporter and receptor in hippocampus and hypothalamus etc. and neuroendocrine system in PSD animal model by the method of neurochemistry, RT-PCT, Western blot, and fMRI, etc. In addition, the relation of depression and hippocampus, hypothalamus etc. will be observed. The effect of JY on the expression of relevant gene in hippocampus, hypothalamus etc. is also assayed at multiple time points by chip technology. The present study aimed to elucidate the characteristics and mechanisms of pharmacological action of JY on PSD, which will provide a theoretical basis and example for Traditional Chinese Medicine (e.g. JY) being used to treat PSD.

中风后抑郁（PSD）是中风后常见的伴随症状。PSD属中医学“郁证”的范畴，肝气郁结为抑郁症的基本病机，“解郁”为其基本治则。临床上，“解郁”法治疗PDS已获得广泛认可。解郁安神颗粒（JY）是部颁标准中药成方制剂，功效为舒肝解郁，安神定志。我们前期的研究显示JY可明显改善局灶性脑缺血大鼠和全脑缺血小鼠的抑郁表现，可调节神经介质5-HT，影响与神经再生相关的BDNF和糖皮质激素受体。但其作用特点及机制尚未明确。本课题以脑缺血后抑郁动物为模型，采用神经化学、细胞和分子生物学、fMRI等技术研究JY对PSD动物不同脑区神经递质、转运体及受体，以及神经-内分泌系统等的影响，观察抑郁与脑区活动的关系，并检测给药后不同时间相关脑区基因和蛋白表达的变化。明确JY “解郁安神”作用的特点，以及经典神经机制和非经典神经网络机制，为JY用于PSD的治疗提供实验依据，为中药治疗PSD的“解郁”治则提供理论支持。

制备PSD动物模型，糖水偏爱实验、强迫游泳实验及悬尾实验观察JY对PSD的治疗作用及其特点；HPLC检测前额叶皮质、海马、纹状体、下丘脑NE，DA及5-HT水平；RT-PCR/Western blot检测不同脑区5-HT1AR、D2R等表达； ELISA检测动物血清ACTH、CORT水平；BrdU/NeuN免疫荧光方法评价马神经再生；分离前额叶皮质、海马、纹状体及下丘脑，RT-PCR/Western blot检测动物不同脑区GR、BDNF等表达；ELISA检测动物血清、前额叶皮质及海马TNF-α、IL-1β、IL-18水平；Western blot检测前额叶皮质及海马NF-κB、IκB-α表达；基因芯片观察海马基因表达谱；MRI及fMRI评价脑结构及功能变化。JY能够明显提高PSD动物的糖水偏爱率，减少不动时间，缩短平衡木行走时间，缩短新奇环境摄食潜伏期，缩短逃避潜伏期，增加目标象限停留距离。表明JY具有抗PSD作用，并可改善PSD伴随的焦虑或认知功能障碍；JY提高 PSD动物海马、下丘脑DA、5-HT及NE水平，上调海马5-HT1AR、D2R、NMDAR1蛋白表达，下调海马ADRα2、GABA-BR蛋白表达。表明JY抗抑郁作用与调节单胺神经递质系统失衡有关；JY降低PSD动物血清ACTH及CORT水平，增加海马齿状回BrdU/NeuN阳性细胞数目，上调海马、前额叶皮质及下丘脑GR、BDNF蛋白表达，下调海马SERT蛋白表达，上调前额叶皮质和纹状体NET1、SERT等蛋白表达，降低血清、前额叶皮质及海马TNF-α、IL-1β及IL-18水平，下调前额叶皮质及海马NF-κB蛋白表达，上调IκB-α蛋白表达。JY诱导海马差异表达基因数目为314个，差异表达基因介导的信号转导通路包括MAPK、NF-κB、Notch、mTOR、神经营养蛋白等。JY改善PSD所致脑组织结构改变，提高海马NAA/Cr，Cho/Cr、mI/Cr比值及前额叶皮质Cho/Cr比值。表明JY抗PSD作用与抑制HPA轴亢进，促进海马神经再生，调节脑GR、BDNF、神经递质转运体蛋白表达，抑制神经炎症及改善脑组织结构、代谢功能异常有关。本课题明确了JY抗PSD作用和特点，以及经典神经机制和非经典神经网络机制，为JY用于PSD的治疗提供实验依据，为中药治疗PSD的“解郁”治则提供理论支持。