School of Laboratory Medicine, Chengdu Medical College, Chengdu, 610500, Sichuan, China.
Department of Orthopedics, Xindu District People's Hospital, Chengdu, 610500, Sichuan, China.
Sci Rep. 2024 Mar 15;14(1):6291. doi: 10.1038/s41598-024-56880-y.
Hedyotis diffusa Willd (HDW) possesses heat-clearing, detoxification, anti-cancer, and anti-inflammatory properties. However, its effects on rheumatoid arthritis (RA) remain under-researched. In this study, we identified potential targets of HDW and collected differentially expressed genes of RA from the GEO dataset GSE77298, leading to the construction of a drug-component-target-disease regulatory network. The intersecting genes underwent GO and KEGG analysis. A PPI protein interaction network was established in the STRING database. Through LASSO, RF, and SVM-RFE algorithms, we identified the core gene MMP9. Subsequent analyses, including ROC, GSEA enrichment, and immune cell infiltration, correlated core genes with RA. mRNA-miRNA-lncRNA regulatory networks were predicted using databases like TargetScan, miRTarBase, miRWalk, starBase, lncBase, and the GEO dataset GSE122616. Experimental verification in RA-FLS cells confirmed HDW's regulatory impact on core genes and their ceRNA expression. We obtained 11 main active ingredients of HDW and 180 corresponding targets, 2150 RA-related genes, and 36 drug-disease intersection targets. The PPI network diagram and three machine learning methods screened to obtain MMP9, and further analysis showed that MMP9 had high diagnostic significance and was significantly correlated with the main infiltrated immune cells, and the molecular docking verification also showed that MMP9 and the main active components of HDW were well combined. Next, we predicted 6 miRNAs and 314 lncRNAs acting on MMP9, and two ceRNA regulatory axes were obtained according to the screening. Cellular assays indicated HDW inhibits RA-FLS cell proliferation and MMP9 protein expression dose-dependently, suggesting HDW might influence RA's progression by regulating the MMP9/miR-204-5p/MIAT axis. This innovative analytical thinking provides guidance and reference for the future research on the ceRNA mechanism of traditional Chinese medicine in the treatment of RA.
白花蛇舌草（HDW）具有清热、解毒、抗癌、抗炎等功效。然而，其在类风湿关节炎（RA）中的作用仍有待研究。在本研究中，我们鉴定了 HDW 的潜在靶点，并从 GEO 数据集 GSE77298 中收集 RA 的差异表达基因，构建了药物-成分-靶标-疾病调控网络。交集基因进行了 GO 和 KEGG 分析。在 STRING 数据库中建立了 PPI 蛋白质相互作用网络。通过 LASSO、RF 和 SVM-RFE 算法，我们鉴定了核心基因 MMP9。随后的分析，包括 ROC、GSEA 富集和免疫细胞浸润，将核心基因与 RA 相关联。使用 TargetScan、miRTarBase、miRWalk、starBase、lncBase 和 GEO 数据集 GSE122616 等数据库预测了 mRNA-miRNA-lncRNA 调控网络。在 RA-FLS 细胞中进行的实验验证证实了 HDW 对核心基因及其 ceRNA 表达的调控作用。我们获得了 11 种 HDW 的主要活性成分和 180 个相应的靶点、2150 个 RA 相关基因和 36 个药物-疾病交集靶点。PPI 网络图和三种机器学习方法筛选获得 MMP9，进一步分析表明 MMP9 具有较高的诊断意义，与主要浸润免疫细胞显著相关，分子对接验证也表明 MMP9 与 HDW 的主要活性成分结合良好。接下来，我们预测了作用于 MMP9 的 6 个 miRNA 和 314 个 lncRNA，并根据筛选结果获得了两个 ceRNA 调控轴。细胞实验表明，HDW 呈剂量依赖性地抑制 RA-FLS 细胞增殖和 MMP9 蛋白表达，提示 HDW 可能通过调节 MMP9/miR-204-5p/MIAT 轴影响 RA 的进展。这种创新的分析思路为今后研究中药 ceRNA 机制在 RA 治疗中的作用提供了指导和参考。
