基于体内药效学结合网络药理学的百合地黄汤抗老年痴呆的机制研究

目的 基于体内药效学结合网络药理学对百合地黄汤治疗老年痴呆的机制进行研究。方法 将清洁级ICR雄性小鼠按体重随机分为6组：正常组、模型组、咯利普兰组、百合地黄汤低、中、高剂量组。连续给药十四天,通过行为学试验来验证百合地黄汤对东莨菪碱诱导的AD模型小鼠行为学变化及药物的改善作用。最后通过呕吐试验来验证百合地黄汤产生呕吐不良反应的程度。通过中药成分数据库 TCMSP、GeneCards、OMIM、DRUGBANK 数据库分别收集百合、地黄化学成分、成分及疾病相关靶点,两者取交集后,运用 STRING 数据库分析靶点蛋白的相互作用,利用Metascape数据库进行生物功能和通路分析,相关结果采用 Cytoscape 3.8.0 进行构图和网络拓扑结构分析,并在此基础上进行机制研究。结果 动物试验结果显示AD小鼠与空白组有显著差异,说明造模成功;百合地黄汤可以逆转AD小鼠的行为学表现,作用与咯利普兰相当。在呕吐试验中,咯利普兰组麻醉苏醒时间显著延长,百合地黄汤组无明显影响,说明其没有呕吐的不良反应。网络药理学中共收集到9个活性成分,54个关键靶点,对接结果显示前10个核心靶点都表现为良好的对接结果。其中对接结果显示其作用与PRKACA、PTGS2密切相关,在此基础上进行了小鼠海马cAMP、PKA、p-CREB和BDNF检测,发现百合地黄汤能纠正东莨菪碱诱导的cAMP-PKA-CREB-BDNF通路下调,作用与阳性药相当,验证了网络药理学的结果。结论 本研究将百合地黄汤对东莨菪碱诱导的AD小鼠模型的抗老年痴呆药效学研究与网络药理学分析结合,预测药物可能通过影响cAMP,上调PTGS2基因的表达以及调控cAMP-PKA-CREB-BDNF信号通路来产生抗老年痴呆作用,为百合地黄汤的临床开发利用提供基础。

Mechanism study of Baihe dihuang decoction on anti-senile dementia based on pharmacodynamics in vivo combined with network pharmacology

Objective The mechanism of Baihe Dihuang decoction in the treatment of senile dementia was studied based on the combination of pharmacodynamics in vivo and network pharmacology. Methods ICR male mice were randomly divided into control group, model group, Rolipram group, low, middle and high dose group of Baihe dihuang decoction. The effect of Baihe Dihuang decoction and the behavior of AD model mice induced by scopolamine was tested by behavior experiment for 14 days. At last, the adverse reaction induced by Baihe dihuang decoction was less than that induced by Rolipram. Chemical constituents of baihe and dihuang, constituents and disease-related targets were collected from TCMSP, GeneCards, OMIM and druggbank databases. respectively, target protein interactions were analyzed with STRING database and biological function and pathway were analyzed with Metascape database.lastly relevant results were analyzed with Cytoscape 3.8.0. Results The results showed that there was significant difference between AD mice and control group, which indicated that the model was successful, and Baihe Dihuang decoction could reverse the behavior of AD mice. In the vomiting experiment, the anesthesia recovery time of the Rolipram group was significantly prolonged, while that of the Baihe Dihuang decoction group was not significantly affected, indicating that Baihe Dihuang decoction was no adverse reaction of vomiting. A total of 9 active components and 54 key targets were collected from network pharmacology. The docking results showed that the first 10 core targets all performed well and their effects were closely related to PRKACA and PTGS2. On this basis, through the detection of cAMP,PKA,p-CREB and BDNF in the hippocampus of mice, it was found that Baihe dihuang decoction could improve the down-regulation of cAMP-PKA-CREB-BDNF pathway induced by scopolamine. Conclusion This study combined the pharmacodynamic study of Baihe Dihuang decoction on AD mice model induced by scopolamine with the analysis of network pharmacology, showing that up-regulation of PTGS2 gene expression and regulation of cAMP-PKA-CREB-BDNF signal pathway in anti-alzheimer"s disease may provide basis for clinical development and utilization of Baihe dihuang decoction.