Key Laboratory of Basic and Application Research of Beiyao (Heilongjiang University of Chinese Medicine), Ministry of Education, Harbin, 150040, China.
Key Laboratory of Basic and Application Research of Beiyao (Heilongjiang University of Chinese Medicine), Ministry of Education, Harbin, 150040, China.
J Ethnopharmacol. 2024 Jun 12;327:118026. doi: 10.1016/j.jep.2024.118026. Epub 2024 Mar 13.
Viscum coloratum (Kom.) Nakai has been traditionally used in China for nearly a thousand years to treat rheumatic diseases. However, its efficacy and mechanisms in treating rheumatoid arthritis (RA) have not been demonstrated.
To investigate the anti-arthritic effects and molecular mechanisms of Viscum coloratum (Kom.) Nakai on collagen-induced arthritic mice through network pharmacology technology and experimental validation.
First, the main ingredients of the extract of Viscum coloratum (Kom.) Nakai (EVC) were identified through chemical composition characterization using Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS). Then, the collagen-induced arthritis (CIA) model was established in DBA/1 J mice and the ameliorative effects of EVC on the progression of CIA mice were evaluated by oral treatment with different doses of the EVC for 28 days. After that, cytokine antibody microarray assay was used to detect the levels of multiple inflammation-related cytokines and chemokines in each group, and performed Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) enrichment analysis. Subsequently, the potential target for the effective chemical components of EVC in treating RA was identified using various databases. Additionally, a drug-disease target protein-protein interaction network (PPI) was conducted using Cytoscape for visualization and clustering, while GO and KEGG enrichment analyses were performed with the Metascape database. Finally, identified phenotypes and targets by network pharmacology analysis were experimentally validated in vivo.
Treatment with EVC significantly suppressed the severity of CIA with a dramatic reduction of paw swelling, arthritis index, levels of IgGs (IgG, IgG1, IgG2a, and IgG2b), multi-inflammation-related cytokines and chemokines on the progression of CIA. Histopathological examinations showed EVC could markedly inhibit inflammatory cell infiltration, tartrate-resistant acid phosphatase (TRAP) activity of osteoclast, and bone destruction. Furthermore, GO and KEGG enrichment analyses revealed that EVC could ameliorate RA by inhibiting osteoclast differentiation and regulating multiple signaling pathways including Osteoclast differentiation, IL-17, and TNF. PPI network analysis demonstrated that AKT1, MMP9, MAPK3, and other genes were highly related to EVC in treating RA. Finally, we proved that EVC could inhibit the expression of NFTAc1, MMP9, Cathepsin K, and AKT which were closely related to osteoclast activity.
EVC could treat RA through multiple components, multiple targets, and multiple pathways. The present study demonstrated the therapeutic efficacy of EVC and its molecular mechanisms in treating RA, indicating that it would be a potent candidate as a novel botanical drug for further investigation.
槲寄生（Kom。）Nakai 在中国传统上已使用近千年治疗风湿性疾病。然而，其在治疗类风湿关节炎（RA）中的功效和机制尚未得到证实。
通过网络药理学技术和实验验证，研究槲寄生（Kom。）Nakai 提取物（EVC）对胶原诱导性关节炎小鼠的抗关节炎作用及其分子机制。
首先，通过超高效液相色谱串联质谱联用（UPLC-MS）对槲寄生（Kom。）Nakai 提取物（EVC）的化学成分进行鉴定。然后，在 DBA/1J 小鼠中建立胶原诱导性关节炎（CIA）模型，并通过口服不同剂量的 EVC 治疗 28 天，评估 EVC 对 CIA 小鼠进展的改善作用。之后，采用细胞因子抗体微阵列检测各组中多种炎症相关细胞因子和趋化因子的水平，并进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。随后，利用多种数据库鉴定 EVC 中有效化学成分治疗 RA 的潜在靶标。此外，使用 Cytoscape 可视化和聚类进行药物-疾病靶蛋白-蛋白相互作用网络（PPI）构建，同时使用 Metascape 数据库进行 GO 和 KEGG 富集分析。最后，通过网络药理学分析鉴定出的表型和靶标在体内进行实验验证。
EVC 治疗可显著抑制 CIA 的严重程度，显著减少爪肿胀、关节炎指数、IgG（IgG、IgG1、IgG2a 和 IgG2b）水平、多种炎症相关细胞因子和趋化因子的产生。组织病理学检查显示，EVC 可明显抑制炎症细胞浸润、抗酒石酸酸性磷酸酶（TRAP）活性的破骨细胞和骨破坏。此外，GO 和 KEGG 富集分析表明，EVC 可通过抑制破骨细胞分化和调节包括破骨细胞分化、IL-17 和 TNF 在内的多种信号通路来改善 RA。PPI 网络分析表明，AKT1、MMP9、MAPK3 等基因与 EVC 治疗 RA 密切相关。最后，我们证明 EVC 可抑制与破骨细胞活性密切相关的 NFTAc1、MMP9、组织蛋白酶 K 和 AKT 的表达。
EVC 可通过多种成分、多个靶点和多条途径治疗 RA。本研究证明了 EVC 治疗 RA 的疗效及其分子机制，表明它可能是一种有前途的新型植物药，值得进一步研究。
