孙雪 梁建庆 何建成 马利芳

【摘 要】 目的：探討敦煌颤震方对PD阴虚动风证大鼠DA能神经元的保护机制。方法：根据大鼠脑立体定位图谱，确定右侧黑质二点坐标，6-OHDA脑立体定位注射法构建PD阴虚动风证模型。采用分层随机法将12只PD阴虚动风证模型大鼠分为2组：模型组，敦煌颤震方组（14 g/kg），每组6只;另取6只大鼠不作造模处理设为正常组。敦煌颤震方组用0.9%氯化钠溶液溶解，每天灌胃1次;正常组和模型组以等体积0.9%氯化钠溶液（2 mL/只）灌胃，连续灌胃6周。处死大鼠，取右侧黑质及纹状体组织，提取总RNA，IlluminaHi Seq平台测序，检测各组大鼠所有基因转录本的差异表达，筛选出与DA能神经元保护机制相关的信号通路及代谢途径。结果：转录数据分析显示：正常组和模型组（右侧黑质/右侧纹状体）之间共筛选出143/149个显著性差异表达基因，其中78/86个基因上调表达，65/63个基因下调表达。经过敦煌颤震方治疗后，差异基因的表达富集程度明显降低， PD模型大鼠PI3K-Akt、MAPK、cAMP、TNF、cGMP-PKG信号通路表达被抑制，氨基酸、蛋白质、维生素B6等代谢途径被激活。结论：敦煌颤震方对PD阴虚动风证大鼠DA能神经元具有一定的保护作用，其机制可能与调节细胞自噬、细胞凋亡、神经细胞再生、神经炎症、氧化应激、突触可塑性、认知功能、氨基酸、蛋白质、维生素B6等信号代谢通路有关。敦煌颤震方可望成为DA能神经元保护剂。

【关键词】 帕金森病;敦煌颤震方;转录组学;多巴胺能神经元

【中图分类号】R285.5   【文献标志码】 A    【文章编号】1007-8517（2021）10-0014-08

Study on the Mechanism of Dunhuang Tremor Prescription Protecting DA Neurons in PD Rats with Yin Deficiency and Moving Wind Syndrome based on Transcriptome

SUN Xue1 LIANG Jianqing1，2* HE Jiancheng3 MA Lifang1

1.School of Basic Medicine， Gansu University of  TCM，Lanzhou 730000， China;

2. The Key Laboratory of the Ministry of Education of Dunhuang medicine and transformation，Lanzhou 730000，China;

3. School of Basic Medicine， Shanghai University of TCM， Shanghai 201203， China

Abstract：Objective To explore the protective mechanism of Dunhuang tremor prescription on DA neurons of PD rats with Yin deficiency and moving wind syndrome.Methods According to the stereotaxic map of the rat brain， the two-point coordinates of the right substantia nigra were determined， and the model of PD Yin deficiency and moving wind syndrome was established by 6-OHDA stereotaxic injection. Twelve PD rats with Yin deficiency and moving wind syndrome were randomly divided into two groups： model group， Dunhuang tremor prescription group （14 g / kg）， 6 rats in each group; another 6 rats were not treated as normal group. Dunhuang tremor prescription group was dissolved with 0.9% sodium chloride solution and administered to the stomach once a day; the normal group and the model group were administered with 0.9% sodium chloride solution of equal volume （2 mL/animal） for 6 weeks. Rats were killed， the right substantia nigra and striatum tissues were taken， total RNA was extracted and sequenced by illuminahi SEQ platform， the differential expression of all gene transcripts in each group was detected， and the signal pathway and metabolic pathway related to the protection mechanism of DA neurons were screened.Results The analysis of transcription data showed that 143 / 149 differentially expressed genes were screened between the normal group and the model group （right substantia nigra / right striatum）， of which 78 / 86 genes were up-regulated and 65 / 63 genes were down regulated. After the treatment of Dunhuang tremor prescription， the expression of differential genes was significantly reduced， the expression of PI3K Akt， MAPK， camp， TNF， cgmp-pkg signaling pathway was inhibited， and the metabolic pathways of amino acid， protein and vitamin B6 were activated.Conclusion Dunhuang tremor prescription has a certain protective effect on DA neurons of PD rats with Yin deficiency and moving wind syndrome. Its mechanism may be related to the regulation of signal metabolic pathways such as autophagy， apoptosis， nerve cell regeneration， neuroinflammation， oxidative stress， synaptic plasticity， cognitive function， amino acid， protein， and vitamin B6. Dunhuang tremor prescription is expected to be a protective agent for DA neurons.

Keywords：Parkinsons Disease; Dunhuang Tremor Prescription;Transcriptome;Dopaminergic Neuron

帕金森病（Parkinsons disease，PD）是全球第二大神经退行性疾病，全世界现有约580万PD患者，中国患病人数约为260万，居世界第一位[1]。……