目的:通过利用网络药理学和分子对接技术,我们可以深入研究益心复麦颗粒(YXFMs)的复杂作用及其对皱纹小鼠病窦综合征(SSS)的影响。具体来说,我们旨在了解YXFM如何通过PI3K/AKT/FOXO途径增强自噬。
方法:人参的活性成分和药用,石竹,麦冬,五味子,丹参,和黄芪使用BATMAN-TCM数据库编制。我们还使用了基因卡,OMIM,和Disgenet文件来确定疾病目标。使用Cytoscape程序生成了“疾病-药物-关键靶标”的分层图。此外,我们使用STRING数据库建立了靶蛋白相互作用(PPI)网络。然后,使用ClusterProfilerR软件包对靶标进行GO功能富集评价和KEGG途径富集分析。基于PPI系统,与分子对接相比,我们选择了最重要的沟通靶标和物质。进行体内研究以进一步验证这些选择。诱导小鼠模型以研究由于年龄相关变化而具有较低心率的小鼠中受损的窦房结(SAN)。进行心电图和Masson染色评估以获得结果。透射电镜观察SAN细胞的自噬水平。采用蛋白质印迹来分析YXFMs对衰老小鼠中整个SSS治疗中PI3K/AKT/FOXO信号传导过程中的蛋白质表达的影响。
结果:一百四十二种活性成分,1858个目标,1226个疾病目标,获得266个相交目标。PPI网络的关键目标包括TP53,AKT1,CTNNB1,INS,和TNF,在其他人中。根据GO功能分析,YXFM在SSS治疗中的潜在机制可能主要与离子跨膜传输的控制有关,心脏收缩,调节血液循环,和其他生物过程。根据KEGG途径富集分析结果,确定它们主要富集在多个信号通路中,如PI3K-Akt信号通路,MAPK信号过程,AGE-RAGE信令路径,FOXO信令路径,HIF-1信令过程,和其他几条路。分子对接表明五种化合物与关键候选靶蛋白AKT1和INS具有良好的结合。通过体内研究,我们注意到服用YXFMs时的显着效果。这些效果包括抑制衰老诱导的SSS,R-R间隔的减少,心率上升,纤维化的减少,自噬过程水平的提高,以及PI3K/AKT/FOXO信号通路中关键蛋白分子的表达水平激增。
结论:本研究对YXFMs治疗SSS的潜力进行了初步预测。这表明YXFM可能具有靶向与病症相关的关键蛋白和关键路径的能力。已经进行了进一步的测试,以发现新的发现和解决老化引发的SSS的想法的证据。
OBJECTIVE: By employing network pharmacology alongside molecular docking techniques, we can delve into the intricate workings of Yixin-Fumai granules (YXFMs) and their impact on sick sinus syndrome (SSS) within wrinkles mice. Specifically, we aim to understand how YXFMs enhance autophagy through the PI3K/AKT/FOXO path.
METHODS: The active ingredients and medicinal uses of Ginseng, ligusticum wallichii, Ophiopogon, Schisandra, salvia, and astragalus were compiled using the BATMAN-TCM database. We also used Genecards, OMIM, and Disgenet files to identify the disease goals. A hierarchical diagram of \"disease-drug-key targets\" was generated using the Cytoscape programs. In addition, we established a target protein interaction (PPI) network using the STRING database. Then, the Cluster Profiler R package was used to conduct GO functional enrichment evaluation and KEGG pathway enrichment analyses of the targets. Based on the PPI system, we chose the top communicating targets and substances over molecular docking. In vivo studies were performed to validate these selections further. The mouse model was induced to study the damaged sinoatrial node (SAN) in mice with lower heart rates due to age-related changes. Electrocardiogram and Masson staining assessments were performed to obtain the results. The transmission electron microscope was used to assess the autophagy level of SAN cells. Western blot was employed to analyze the impact of YXFMs on protein expression in the PI3K/AKT/FOXO signaling process throughout SSS therapy in aging mice.
RESULTS: One hundred forty-two active ingredients, 1858 targets, 1226 disease targets, and 266 intersection targets were obtained. The key targets of the PPI network encompassed TP53, AKT1, CTNNB1, INS, and TNF, among others. According to GO functional analysis, the mechanism underlying YXFMs in SSS treatment may primarily be associated with the control of ion transport across membranes, cardiac contraction, regulation of blood circulation, and other biological processes. Based on the results of KEGG pathway enrichment analysis, it was determined that they were mainly enriched in multiple pathways of signaling such as the PI3K-Akt signaling route, MAPK signaling process, AGE-RAGE signaling path, FOXO signaling path, HIF-1 signaling process, and several other paths. Molecular docking demonstrated that five compounds had excellent binding to the key candidate target proteins AKT1 and INS. Through the in vivo studies, we noticed notable effects when administering YXFMs. These effects included the suppression of aging-induced SSS, a decrease in the R-R interval, a rise in heart rate, a reduction in fibrosis, a boost in the autophagy process level, and a spike in the levels of expression of key protein molecules in the PI3K/AKT/FOXO signaling path.
CONCLUSIONS: This research has made preliminary predictions about the potential of YXFMs in treating SSS. It suggests that YXFMs may have the ability to target key proteins and critical paths associated with the condition. Further testing has been conducted to discover new findings and evidence of ideas for tackling SSS triggered by aging.