运用网络药理学、分子对接和实验验证的方法揭示地黄治疗银屑病的作用机制。
Integrating network pharmacology, molecular docking, and experimental validation to reveal the mechanism of Radix Rehmanniae in psoriasis.
机构信息
The Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, China.
State Key Laboratory of TCM Moisture Syndrome at the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China.
出版信息
Medicine (Baltimore). 2024 Oct 25;103(43):e40211. doi: 10.1097/MD.0000000000040211.
BACKGROUND
Radix Rehmanniae (RR) plays an important role in treating psoriasis. However, the active compounds of RR and potential mechanisms are unclear. The current study was designed to investigate the potential active ingredients, targets, and mechanisms of RR in treating psoriasis through network pharmacology, molecular docking, and vitro experiments.
METHODS
Initially, the TCMSP database and literature retrieval were used to access the active ingredients of RR. The psoriasis target proteins were obtained from Therapeutic Target Database, OMIM, GeneCards, and DrugBank databases. The target proteins were then converted into target genes using Uniprot. Secondly, overlapping genes were obtained through Venn online tool. Then, protein-protein interactions network diagram is finished by STRING database. Next, Cytoscape software was used to acquire the top 10 hub proteins; gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis were then used to predict possible mechanisms. Afterwards, molecular docking validation of the active ingredients with the main targets was performed by AutoDock software. Finally, lipopolysaccharides induced RAW264.7, to assess the effects and molecular mechanisms by MTT, RT-qPCR, and Western blot assays.
RESULTS
Overall, there are 20 effective compounds and 33 targets involved in biological processes including apoptosis, intracellular signaling, vasodilation, and mitogen-activated protein kinase (MAPK) signaling cascade. The docking results showed strong binding capacity between the active ingredients and targets. We verified aucubin as the key active ingredient, tumor necrosis factor α, and IL6 as the core targets, and focused on the p38MAPK protein pathway. Cellular experiments showed that aucubin down-regulated the phosphorylated p38MAP protein and reduced the expression of tumor necrosis factor α mRNA, IL6 mRNA, and IL1βmRNA.
CONCLUSION
In summary, RR is featured with multicomponent, multi-target, and multi-pathway in treating psoriasis; the preliminary mechanism may be associated with the down-regulation of p38MAPK phosphorylation and curbing the expression of inflammatory factor by aucubin. This paper provides the scientific basis for Traditional Chinese medicine treating psoriasis.
背景
地黄在治疗银屑病中起着重要作用。然而，地黄的活性化合物和潜在机制尚不清楚。本研究旨在通过网络药理学、分子对接和体外实验研究地黄治疗银屑病的潜在活性成分、靶点和机制。
方法
首先，利用 TCMSP 数据库和文献检索获取地黄的活性成分。从治疗靶点数据库、OMIM、基因卡片和药物银行数据库中获得银屑病靶点蛋白。然后，使用 Uniprot 将靶点蛋白转化为靶点基因。其次，通过 Venn 在线工具获得重叠基因。然后，通过 STRING 数据库完成蛋白质-蛋白质相互作用网络图。接下来，使用 Cytoscape 软件获取前 10 个枢纽蛋白；然后进行基因本体论和京都基因与基因组百科全书富集分析，以预测可能的机制。之后，使用 AutoDock 软件对主要靶点的活性成分进行分子对接验证。最后，通过 MTT、RT-qPCR 和 Western blot 测定法评估脂多糖诱导的 RAW264.7 细胞的作用和分子机制。
结果
总的来说，有 20 种有效化合物和 33 个靶点参与了包括细胞凋亡、细胞内信号转导、血管扩张和丝裂原激活蛋白激酶（MAPK）信号级联在内的生物学过程。对接结果表明，活性成分与靶点之间具有很强的结合能力。我们验证了梓醇是关键的活性成分，肿瘤坏死因子-α和白细胞介素 6 是核心靶点，并关注了 p38MAPK 蛋白通路。细胞实验表明，梓醇下调磷酸化 p38MAP 蛋白的表达，降低肿瘤坏死因子-αmRNA、白细胞介素 6mRNA 和白细胞介素 1βmRNA 的表达。
结论
综上所述，地黄在治疗银屑病方面具有多成分、多靶点和多途径的特点；初步机制可能与 p38MAPK 磷酸化的下调和梓醇抑制炎症因子的表达有关。本文为中药治疗银屑病提供了科学依据。
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