Liu Zengying, Guan Chen, Li Chenyu, Zhang Ningxin, Yang Chengyu, Xu Lingyu, Zhou Bin, Zhao Long, Luan Hong, Man Xiaofei, Xu Yan
Department of Nephrology, the Affiliated Hospital of Qingdao University, Qingdao, China.
Medizinische Klinik und Poliklinik IV, Klinikum der Universität, LMU München, München, Germany.
Front Pharmacol. 2022 Jun 3;13:862584. doi: 10.3389/fphar.2022.862584. eCollection 2022.
Acute kidney injury (AKI) is a common syndrome impacting about 13.3 million patients per year. Tilianin has been reported to alleviate myocardial ischemia/reperfusion (I/R) injury, while its effect on AKI is unknown; thus, this study aimed to explore if tilianin protects I/R-induced AKI and the underlying mechanisms. The microarray dataset GSE52004 was downloaded from GEO DataSets (Gene Expression Omnibus). Differential expression analysis and gene-set enrichment analysis (GSEA) were performed by R software to identify apoptosis pathway-related genes. Then, was applied to identify the transcription factor (TF) related to apoptosis. The STRING database was used to construct a protein-protein interaction (PPI) network. Cytoscape software visualized PPI networks, and hub TFs were selected via AutoDock was used for molecular docking of tilianin and hub gene-encoded proteins. The expression levels of hub genes were assayed and visualized by quantitative real-time PCR, Western blotting, and immunohistochemistry by establishing I/R-induced AKI mouse models. Bioinformatics analysis showed that 34 genes, including FOS, ATF4, and Gadd45g, were involved in the apoptosis pathway. In total, seven hub TFs might play important roles in tilianin-regulating apoptosis pathways. In , tilianin improved kidney function and reduced the number of TUNEL-positive renal tubular epithelial cells (RTECs) after I/R-induced AKI. Tilianin reduced the activation of the ERK pathway and then downregulated the expression of EGR1. This further ameliorated the expression of anti-apoptotic genes such as BCL2L1 and BCL2, reduced pro-apoptotic genes such as BAD, BAX, and caspase-3, and reduced the release of cytochrome c. Tilianin reduced apoptosis after I/R-induced AKI by the ERK/EGR1/BCL2L1 pathway. Our findings provided novel insights for the first time into the protective effect and underlying molecular mechanisms of tilianin on I/R-induced AKI.
急性肾损伤（AKI）是一种常见综合征，每年影响约1330万患者。据报道，田基黄苷可减轻心肌缺血/再灌注（I/R）损伤，但其对AKI的影响尚不清楚；因此，本研究旨在探讨田基黄苷是否能保护I/R诱导的AKI及其潜在机制。从基因表达综合数据库（GEO DataSets）下载微阵列数据集GSE52004。通过R软件进行差异表达分析和基因集富集分析（GSEA），以鉴定凋亡途径相关基因。然后，应用该软件鉴定与凋亡相关的转录因子（TF）。利用STRING数据库构建蛋白质-蛋白质相互作用（PPI）网络。Cytoscape软件可视化PPI网络，并通过自动对接选择枢纽转录因子。通过建立I/R诱导的AKI小鼠模型，采用定量实时PCR、蛋白质印迹法和免疫组织化学法检测并可视化枢纽基因的表达水平。生物信息学分析表明，包括FOS、ATF4和Gadd45g在内的34个基因参与了凋亡途径。总共有7个枢纽转录因子可能在田基黄苷调节凋亡途径中发挥重要作用。在体内，田基黄苷改善了I/R诱导的AKI后的肾功能，并减少了TUNEL阳性肾小管上皮细胞（RTECs）的数量。田基黄苷降低了ERK途径的激活，进而下调了EGR1的表达。这进一步改善了抗凋亡基因如BCL2L1和BCL2的表达，降低了促凋亡基因如BAD、BAX和caspase-3的表达，并减少了细胞色素c的释放。田基黄苷通过ERK/EGR1/BCL2L1途径减少了I/R诱导的AKI后的细胞凋亡。我们的研究结果首次为田基黄苷对I/R诱导的AKI的保护作用及其潜在分子机制提供了新的见解。
