Exploration of the Effect and Mechanism of Fructus Lycii, Rehmanniae Radix Praeparata, and Paeonia lactiflora in the Treatment of AMD Based on Network Pharmacology and in vitro Experimental Verification.
Exploration of the Effect and Mechanism of Fructus Lycii, Rehmanniae Radix Praeparata, and Paeonia lactiflora in the Treatment of AMD Based on Network Pharmacology and in vitro Experimental Verification.
复制标题
基于网络药理学及体外实验验证枸杞子、熟地、芍药治疗AMD的作用及机制探讨
DOI：
10.2147/dddt.s310481
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Wei W
中科院分区：
文献类型：
--
作者：
Chen X;Zuo J;Hu T;Shi X;Zhu Y;Wu H;Xia Y;Shi W;Wei W
目的以网络药理学和生物学实验为基础，观察枸杞子、地黄、白药治疗老年性黄斑变性(AMD)的作用机制。方法从公共数据库中获取生物活性物质、FL、RRP和PL的潜在靶点以及与AMD相关的基因。基因本体论(GO)和京都基因和基因组百科全书(KEGG)对核心靶标进行了功能和途径浓缩分析。随后，这一发现得到了细胞实验的进一步验证。采用四甲基偶氮唑盐比色法和流式细胞仪检测细胞存活率和细胞凋亡率。用DCFH-DA染色分析ROS的产生；用试剂盒化学测定抗氧化酶的活性。用免疫印迹法检测关键蛋白的表达。结果共鉴定出59个活性化合物、182个潜在靶点和2536个AMD相关人类基因。通过蛋白质-蛋白质相互作用(PPI)分析，确定了三种中药治疗AMD的103个关键靶点。根据GO和KEGG分析，上述靶点与核受体活性、氧化应激和细胞凋亡途径相关。四甲基偶氮唑盐比色法和流式细胞仪检测结果表明，三味中药可显著提高ARPE-19细胞的存活率，减少过氧化氢诱导的细胞凋亡。三味中药均可降低过氧化氢后细胞内ROS水平，提高超氧化物歧化酶和过氧化氢酶的活性。此外，这三种中药还通过增加Nrf2、HO-1和NQO1的表达而显著抑制氧化应激。结论网络药理实验和验证实验相结合的结果表明，FL、RRP和PL可降低RPE细胞的氧化应激和细胞凋亡，从而发挥其治疗AMD的作用。
Purpose The aim of this study was to observe the mechanism of Fructus Lycii (FL), Rehmanniae Radix Praeparata (RRP) and Paeonia lactiflora (PL) in treating age-related macular degeneration (AMD) based on network pharmacology and biological experiments. Methods Bioactive compounds, potential targets of FL, RRP and PL, and genes related to AMD, were acquired from public databases. Functional and pathway enrichment analyses of the core targets were conducted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Subsequently, the finding was further verified with cell experiments. The MTT assay and flow cytometric analysis were used to assess cell viability and apoptosis. The production of reactive oxygen species (ROS) was analyzed by DCFH-DA staining; the activity of antioxidant enzymes was chemically measured with assay kits. The expression of key proteins was evaluated by Western blot analysis. Results Fifty-nine active compounds, 182 potential targets, and 2536 AMD-related human genes were identified. A total of 103 key targets of the three herbs on AMD were identified by protein–protein interaction (PPI) analysis. The abovementioned targets were correlated with nuclear receptor activity, oxidative stress, and apoptosis pathways according to the GO and KEGG analyses. MTT assay and flow cytometry demonstrated that pretreatment of ARPE-19 cells with the three herbs significantly increased cell viability and decreased apoptosis induced by H2O2. The three herbs might reduce the intracellular ROS levels and increase the SOD and CAT activities after H2O2. Furthermore, the three herbs significantly inhibited oxidative stress via increasing the expression of Nrf2, HO-1 and NQO1. Conclusion The combined results of network pharmacology and validation experiments showed that FL, RRP and PL reduce oxidative stress and apoptosis in RPE cells to exert its effect in the treatment of AMD.