背景:红景天苷(SAL),红景天提取物的主要成分,是一种具有生物活性的类黄酮,如抗氧化应激,抗炎,和降血脂。在这项研究中,在体外实验的基础上,研究了SAL抗视网膜神经节细胞(RGCs)氧化应激的潜在治疗靶点和机制,网络药理学,和分子对接技术。
方法:构建RGC氧化应激模型,和细胞活动,活性氧(ROS),并检查细胞凋亡水平的差异。对应于视紫红质的基因,RGC,从GeneCards中筛选出氧化应激,TCMSP数据库,和一个分析平台。三人的交叉点被拿走了,绘制了维恩图。蛋白质相互作用,GO功能富集,和KEGG途径富集数据通过STRING数据库进行分析,Cytohubba插件,和Metascape数据库。使用qRT-PCR验证筛选途径中的关键因素。最后,使用MOE2019软件进行分子对接预测,使用Gromacs2018软件进行分子动力学模拟。
结果:在体外RGC氧化应激模型中,细胞活性增强,ROS减少了,SAL处理后细胞凋亡减少。获得了SALRGCs中16个潜在的氧化应激目标,并通过网络拓扑分析筛选前10名核心目标。GO分析表明,SAL视网膜氧化应激治疗主要涉及细胞对应激反应,转录调控复合物,和DNA结合转录因子结合。KEGG分析显示,大多数基因主要富集在糖尿病并发症的多个肿瘤通路和信号通路中,非酒精性脂肪肝,和脂质和动脉粥样硬化。通过PCR验证,分子对接和分子动力学模拟显示SAL可能通过调节SIRT1、NRF2和NOS3减弱RGCs的氧化应激和减少细胞凋亡。
结论:本研究初步揭示了SAL对RGCs的抗氧化治疗作用和分子机制,为后续研究提供理论依据。
BACKGROUND: Salidroside (SAL), the main component of Rhodiola rosea extract, is a flavonoid with biological activities, such as antioxidative stress, anti-inflammatory, and hypolipidemic. In this study, the potential therapeutic targets and mechanisms of SAL against oxidative stress in retinal ganglion cells (RGCs) were investigated on the basis of in-vitro experiments, network pharmacology, and molecular docking techniques.
METHODS: RGC oxidative stress models were constructed, and cell activity, reactive oxygen species (ROS), and apoptosis levels were examined for differences. The genes corresponding to rhodopsin, RGCs, and oxidative stress were screened from GeneCards, TCMSP database, and an analysis platform. The intersection of the three was taken, and a Venn diagram was drawn. Protein interactions, GO functional enrichment, and KEGG pathway enrichment data were analyzed by STRING database, Cytohubba plugin, and Metascape database. The key factors in the screening pathway were validated using qRT-PCR. Finally, molecular docking prediction was performed using MOE 2019 software, molecular dynamic simulations was performed using Gromacs 2018 software.
RESULTS: In the RGC oxidative stress model in vitro, the cell activity was enhanced, ROS was reduced, and apoptosis was decreased after SAL treatment. A total of 16 potential targets of oxidative stress in SAL RGCs were obtained, and the top 10 core targets were screened by network topology analysis. GO analysis showed that SAL retinal oxidative stress treatment mainly involved cellular response to stress, transcriptional regulatory complexes, and DNA-binding transcription factor binding. KEGG analysis showed that most genes were mainly enriched in multiple cancer pathways and signaling pathways in diabetic complications, nonalcoholic fatty liver, and lipid and atherosclerosis. Validation by PCR, molecular docking and molecular dynamic simulations revealed that SAL may attenuate oxidative stress and reduce apoptosis in RGCs by regulating SIRT1, NRF2, and NOS3.
CONCLUSIONS: This study initially revealed the antioxidant therapeutic effects and molecular mechanisms of SAL on RGCs, providing a theoretical basis for subsequent studies.