DataSheet3_Pantao Pill Improves the Learning and Memory Abilities of APP/PS1 Mice by Multiple Mechanisms.zip

Background: To explore the effect and mechanisms of Pantao Pill (PTP) on cognitive impairment.
Methods: Network pharmacology was performed to analyze the mechanism of PTP treating cognitive impairment. The targets of PTP and cognitive impairment were predicted and used to construct protein-protein interaction (PPI) networks. The intersection network was selected, and the core network was obtained through topological analysis. Enrichment analysis was conducted to obtain the GOBP terms and KEGG pathways. We then performed experiments to validate the results of the network pharmacology by using an APP/PS1 transgenic mouse model. The APP/PS1 mice were divided into four groups: the model group, the high-dose PTP (3.6 g/kg·d) group, the low-dose PTP (1.8 g/kg·d) group, and the positive control group (donepezil hydrochloride, 2 mg/kg·d). Wild-type (WT) C57 mice served as a normal control group. PTP and donepezil were administered by gavage for 8 weeks.
Results: Network pharmacology showed that PTP might improve cognitive impairment by regulating autophagy, apoptosis, and oxidative stress. For the Morris water maze test, a significant difference was shown in the total swimming distance among groups (p < 0.05) in the positioning navigation experiment, and with training time extension, the swimming speed increased (p < 0.01). In the space probe test, PTP administration significantly reduced the swimming path length and the escape latency of APP/PS1 mice (p < 0.05 or p < 0.01), whereas it had no effect on the swimming speed (p > 0.05). PTP (3.6 g/kg/d) rescued the reduction of norepinephrine and acetylcholine levels (p < 0.05), and increased the acetylcholinesterase concentration (p < 0.05) in the brain tissue. PTP (1.8 g/kg/d) increased the norepinephrine level (p < 0.01). PTP rescued the activity reduction of superoxide dismutase in the brain tissue (p < 0.01) and the neuron cell pyknosis in the hippocampal CA region (p < 0.05). PTP reduced ATG12 and PS1 expression (p < 0.05 or p < 0.01), and increased Bcl-2 expression in the brain tissue (p < 0.05).
Conclusion: PTP can significantly improve the learning and memory abilities of APP/PS1 mice, and the mechanism may be related to the increase of neurotransmitter acetylcholine and norepinephrine levels, the reduction of the excessive autophagic activation, and the suppression of oxidative stress and excessive apoptotic activity.

背景：为探讨瘫桃丸（Pantao Pill, PTP）对认知障碍的作用及其机制。方法：采用网络药理学方法分析PTP治疗认知障碍的作用机制。预测PTP与认知障碍的作用靶点，并构建蛋白-蛋白相互作用（protein-protein interaction, PPI）网络。选取交集网络，通过拓扑分析获取核心网络。开展富集分析以获得基因本体生物过程（Gene Ontology Biological Process, GOBP）术语及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路。随后采用APP/PS1转基因小鼠模型对网络药理学分析结果进行实验验证。将APP/PS1小鼠分为4组：模型组、高剂量PTP组（3.6 g/kg·d）、低剂量PTP组（1.8 g/kg·d）及阳性对照组（盐酸多奈哌齐，2 mg/kg·d）；以野生型（Wild-type, WT）C57小鼠作为正常对照组。采用灌胃方式给予PTP与盐酸多奈哌齐，持续给药8周。结果：网络药理学分析显示，PTP或可通过调控自噬、细胞凋亡及氧化应激改善认知障碍。莫里斯水迷宫（Morris water maze）定位导航实验中，各组小鼠总游泳距离存在显著组间差异（p < 0.05）；随着训练时长增加，小鼠游泳速度显著提升（p < 0.01）。空间探索实验中，PTP给药可显著缩短APP/PS1小鼠的游泳路径长度与逃避潜伏期（p < 0.05 或 p < 0.01），但对游泳速度无显著影响（p > 0.05）。3.6 g/kg/d剂量的PTP可恢复脑组织中去甲肾上腺素与乙酰胆碱水平的降低（p < 0.05），并提升脑组织中乙酰胆碱酯酶浓度（p < 0.05）；1.8 g/kg/d剂量的PTP可提升去甲肾上腺素水平（p < 0.01）。PTP可逆转脑组织中超氧化物歧化酶活性的降低（p < 0.01），并改善海马CA区神经元固缩现象（p < 0.05）。PTP可降低脑组织中ATG12与PS1的表达水平（p < 0.05 或 p < 0.01），并提升Bcl-2的表达水平（p < 0.05）。结论：PTP可显著改善APP/PS1小鼠的学习记忆能力，其潜在机制可能与提升神经递质乙酰胆碱与去甲肾上腺素水平、抑制过度自噬激活、减轻氧化应激及过度细胞凋亡活性相关。