Department of Internal Medicine, Henan Medical College, Zhengzhou, China.
Academy of Chinese Medicine Sciences, Henan University of Chinese Medicine, Zhengzhou, China.
Gene. 2025 Jan 20;934:149056. doi: 10.1016/j.gene.2024.149056. Epub 2024 Oct 28.
Renal fibrosis is a crucial factor in the progression of chronic kidney diseases. Previous studies have suggested that apigenin (API) has potential in ameliorating renal fibrosis, but its therapeutic mechanism remains unclear. This study aims to elucidate the mechanisms by which API treats renal fibrosis using network pharmacology and experimental validation. Initially, we used the Traditional Chinese Medicine Systems Pharmacology (TCMSP) Database and GeneCards database to identify molecular targets of API and associated genes. Next, we constructed a network of API-renal fibrosis targets, followed by protein-protein interaction (PPI) analysis. Subsequent analyses, such as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, were performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID). We also performed molecular docking studies to explore API's interactions with key proteins. To validate API's mechanism in treating renal fibrosis, we used a Human Kidney-2 (HK-2) cell model of epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1). We identified 77 API target genes, 8434 renal fibrosis target genes, and 64 intersection genes, which were primarily enriched in nuclear factor kappa-B (NF-κB) and Phosphatidylinositide 3-kinases/protein kinase B (PI3K-AKT) pathways. API significantly inhibited EMT in TGF-β1-induced HK-2 cells by regulating the expression of α-Smooth muscle actin (α-SMA) and E-cadherin and suppressing the protein expression of p-PI3K, p-AKT, and p-P65, which are related to the PI3K-AKT and NF-κB pathways. However, co-administration of the PI3K agonist 740Y-P counteracted API's inhibitory effects on these protein expressions. In summary, these findings highlight API's therapeutic potential in treating renal fibrosis by modulating EMT in renal tubular epithelial cells via the PI3K-AKT and NF-κB pathways.
肾纤维化是慢性肾脏病进展的关键因素。先前的研究表明，芹菜素（API）在改善肾纤维化方面具有潜力，但其治疗机制尚不清楚。本研究旨在通过网络药理学和实验验证来阐明 API 治疗肾纤维化的机制。首先，我们使用中药系统药理学（TCMSP）数据库和基因卡片数据库来识别 API 的分子靶标和相关基因。接下来，我们构建了 API-肾纤维化靶标网络，随后进行了蛋白质-蛋白质相互作用（PPI）分析。随后，使用数据库进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析（DAVID）。我们还进行了分子对接研究，以探索 API 与关键蛋白的相互作用。为了验证 API 治疗肾纤维化的机制，我们使用了由转化生长因子-β1（TGF-β1）诱导的人肾-2（HK-2）细胞上皮-间充质转化（EMT）模型。我们鉴定了 77 个 API 靶基因、8434 个肾纤维化靶基因和 64 个交集基因，这些基因主要富集在核因子 kappa-B（NF-κB）和磷脂酰肌醇 3-激酶/蛋白激酶 B（PI3K-AKT）通路中。API 通过调节α-平滑肌肌动蛋白（α-SMA）和 E-钙黏蛋白的表达以及抑制与 PI3K-AKT 和 NF-κB 通路相关的 p-PI3K、p-AKT 和 p-P65 的蛋白表达，显著抑制了 TGF-β1 诱导的 HK-2 细胞中的 EMT。然而，PI3K 激动剂 740Y-P 的共同给药抵消了 API 对这些蛋白表达的抑制作用。总之，这些发现强调了 API 通过调节 PI3K-AKT 和 NF-κB 通路在肾小管上皮细胞 EMT 中的作用，从而具有治疗肾纤维化的治疗潜力。
