Department of Oral Medicine, Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan 030001, China.
Department of Physiology, School of Medicine, China Medical University, Taichung 40402, Taiwan.
Phytomedicine. 2022 Aug;103:154195. doi: 10.1016/j.phymed.2022.154195. Epub 2022 May 22.
Oral leukoplakia (OLK), an uncharacterized pathological condition that occurs as a white patch in the oral mucosa, is the most common precancerous condition. Scutellaria baicalensis Georgi (SBG) is a medicinal plant with a wide range of pharmacological effects. Increased evidence shows that SBG has potential therapeutic effects on OLK. However, the therapeutic mechanisms of SBG against OLK have not yet been completely elucidated.
This study aimed to clarify the active components and multi-target mechanisms of SBG against OLK via network pharmacology, molecular docking and experimental evaluations.
The active components and related targets of SBG were screened by the TCMSP database and Swiss Target Prediction database. Potential therapeutic targets of OLK were collected using the GeneCards and OMIM databases. Then, we established protein-protein interaction (PPI), compound-target-disease (C-T-D), and compound-target-pathway (C-T-P) networks by Cytoscape to identify the main components, core targets, and pharmacological pathways of SBG against OLK via applying data mining techniques and topological parameters. Metascape database was utilized for GO and KEGG pathway analysis. Molecular docking techniques were used to estimate the binding force between the components and the hub genes. Subsequently, a series of in vitro experiments, specifically CCK-8 assay, clone formation assay, wound healing assay, flow cytometry, RT-qPCR and western blotting were conducted for further verification.
There were 25 active components and 31 related target genes in SBG against OLK. PPI analysis showed that Akt1, VEGFA, EGFR, HIF1A and PTGS2 shared the highest centrality among all target genes. KEGG pathway analysis found that PI3K-Akt signaling pathway may occupy core status in the anti-OLK system. Molecular docking results showed that the main active components of SBG had a strong binding affinity to the hub genes. In vitro experiments showed that the leading component baicalein may inhibit proliferation, block cells in the S phase, induce DOK cell apoptosis, and downregulate the mRNA expression of 5 hub genes by inhibiting PI3K/Akt signaling pathway activation.
The most predominant component of SBG against OLK was baicalein and the key pathway was PI3K/Akt. The main components and hub genes had robust binding abilities. In vitro experiments showed that baicalein could inhibit the proliferation of DOK cells, induce apoptosis, block the cell cycle, and inhibit the mRNA expression level of the hub genes by inhibiting the PI3K/Akt pathway.
口腔白斑病（OLK）是一种口腔黏膜出现白色斑块的特征不明的病理状态，是最常见的癌前病变。黄芩（SBG）是一种具有广泛药理作用的药用植物。越来越多的证据表明，黄芩对 OLE 具有潜在的治疗作用。然而，黄芩治疗 OLE 的机制尚未完全阐明。
本研究旨在通过网络药理学、分子对接和实验评价，阐明 SBG 对 OLE 的活性成分和多靶点机制。
利用 TCMSP 数据库和瑞士靶点预测数据库筛选 SBG 的活性成分和相关靶点。利用 GeneCards 和 OMIM 数据库收集 OLE 的潜在治疗靶点。然后，我们通过 Cytoscape 建立蛋白质-蛋白质相互作用（PPI）、化合物-靶点-疾病（C-T-D）和化合物-靶点-通路（C-T-P）网络，通过数据挖掘技术和拓扑参数识别 SBG 治疗 OLE 的主要成分、核心靶点和药理途径。利用 Metascape 数据库进行 GO 和 KEGG 通路分析。分子对接技术用于估计成分与枢纽基因之间的结合力。随后，进行了一系列体外实验，包括 CCK-8 测定、克隆形成测定、划痕愈合测定、流式细胞术、RT-qPCR 和 Western blot，以进一步验证。
SBG 治疗 OLE 有 25 个活性成分和 31 个相关靶点。PPI 分析表明，在所有靶点基因中，Akt1、VEGFA、EGFR、HIF1A 和 PTGS2 具有最高的中心性。KEGG 通路分析发现，PI3K-Akt 信号通路可能在抗 OLE 系统中占据核心地位。分子对接结果表明，SBG 的主要活性成分与枢纽基因具有很强的结合亲和力。体外实验表明，黄芩的主要成分白杨素可能通过抑制 PI3K/Akt 信号通路的激活来抑制细胞增殖、阻断细胞进入 S 期、诱导 DOK 细胞凋亡，并下调 5 个枢纽基因的 mRNA 表达。
SBG 治疗 OLE 的最主要成分是白杨素，关键通路是 PI3K/Akt。主要成分和枢纽基因具有很强的结合能力。体外实验表明，白杨素通过抑制 PI3K/Akt 通路，抑制 DOK 细胞增殖，诱导凋亡，阻断细胞周期，抑制枢纽基因的 mRNA 表达水平，从而抑制 DOK 细胞的增殖。
