Bai Li, Li Fang
The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
Bazhong Central Hospital, Bazhong, China.
Medicine (Baltimore). 2024 Dec 6;103(49):e40550. doi: 10.1097/MD.0000000000040550.
This study aims to investigate the effect of promethazine (PMZ) on hippocampal neuronal injury through network pharmacology and in vivo experiments. Network pharmacology: The intersection genes of PMZ and Alzheimer Disease (AD) were obtained, and the core genes of PMZ in AD were screened. The intersection genes were enriched by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. In the in vitro experiment, mouse hippocampal neurons (HT22) were divided into control, glutamate (GLU) model, and GLU + PMZ treatment groups. The control group was given a complete culture medium, the model group was given GLU for 24 hours, the treatment group was given PMZ pretreatment for 3 hours, and then GLU was administered for 24 hours. Cell viability was determined, cell morphology was observed by microscopy, reactive oxygen species levels and glutathione content were detected, and protein expression of P53, PTGS2, SLC7A11, and GPX4 was detected by western blotting. Network pharmacology: A total of 317 PMZ targets, 1934 AD genes, 125 intersection genes, and 18 core genes, including P53 and PTGS2. Gene Ontology enrichment analysis showed that the effect of PMZ on AD was mainly related to cell proliferation, inflammation, hypoxia, synaptic structure, plasma membrane, and oxidoreductase activity. Kyoto Encyclopedia of Genes and Genomes results showed neuroactive ligand-receptor interaction, cell senescence, cancer pathway, PI3K-AKT signal pathway, neurodegeneration, and HIF-1 signal pathway. In vitro experiments: PMZ improved the GLU-induced decrease in cell viability and morphological changes in hippocampal neurons. PMZ inhibited reactive oxygen species levels and increased glutathione content in injured hippocampal neurons. Up-regulated of P53, SLC7A11 and GPX4 expression, and inhibited expression of PTGS2. PMZ regulates the SLC7A11-GPX4 antioxidant system to protect hippocampal neurons from oxidative stress injury.
本研究旨在通过网络药理学和体内实验探讨异丙嗪（PMZ）对海马神经元损伤的影响。网络药理学：获取PMZ与阿尔茨海默病（AD）的交集基因，筛选出PMZ在AD中的核心基因。通过基因本体论（Gene Ontology）和京都基因与基因组百科全书（KEGG）通路富集分析对交集基因进行富集。在体外实验中，将小鼠海马神经元（HT22）分为对照组、谷氨酸（GLU）模型组和GLU + PMZ治疗组。对照组给予完全培养基，模型组给予GLU 24小时，治疗组给予PMZ预处理3小时，然后给予GLU 24小时。测定细胞活力，通过显微镜观察细胞形态，检测活性氧水平和谷胱甘肽含量，并通过蛋白质免疫印迹法检测P53、PTGS2、SLC7A11和GPX4的蛋白表达。网络药理学：共获得317个PMZ靶点、1934个AD基因、125个交集基因和18个核心基因，包括P53和PTGS2。基因本体论富集分析表明，PMZ对AD的作用主要与细胞增殖、炎症、缺氧、突触结构、质膜和氧化还原酶活性有关。KEGG结果显示神经活性配体-受体相互作用、细胞衰老、癌症通路、PI3K-AKT信号通路、神经退行性变和HIF-1信号通路。体外实验：PMZ改善了GLU诱导的海马神经元细胞活力下降和形态变化。PMZ抑制了损伤海马神经元中的活性氧水平并增加了谷胱甘肽含量。上调P53、SLC7A11和GPX4的表达，并抑制PTGS2的表达。PMZ调节SLC7A11-GPX4抗氧化系统以保护海马神经元免受氧化应激损伤。
