Table5_Niaoduqing alleviates podocyte injury in high glucose model via regulating multiple targets and AGE/RAGE pathway: Network pharmacology and experimental validation.pdf

Purpose: The aim of present study was to explore the pharmacological mechanisms of Niaoduqing granules on the treatment of podocyte injury in diabetic nephropathy (DN) via network pharmacology and experimental validation.
Methods: Active ingredients and related targets of Niaoduqing, as well as related genes of podocyte injury, proteinuria and DN, were obtained from public databases. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction (PPI) network analysis were performed to investigate the potential mechanisms. High glucose (HG) -induced MPC5 cell injury model was treated with the major core active ingredients of Niaoduqing and used to validate the predicted targets and signaling pathways.
Results: Totally, 16 potential therapeutic targets were identified by intersecting the targets of Niaoduqing and disease, in which 7 of them were considered as the core targets via PPI network analysis. KEGG enrichment analysis showed that AGE-RAGE signaling pathway was identified as the most crucial signaling pathway. The results of in vitro experiments revealed that the treatment of Niaoduqing active ingredients significantly protected MPC5 cells from HG-induced apoptosis. Moreover, Niaoduqing could significantly attenuate the HG-induced activation of AGE-RAGE signaling pathway, whereas inhibited the over-expression of VEGF-A, ICAM-1, PTGS-2 and ACE in HG-induced MPC5 cells.
Conclusion: Niaoduqing might protect against podocyte injury in DN through regulating the activity of AGE/RAGE pathway and expression of multiple genes. Further clinical and animal experimental studies are necessary to confirm present findings.

研究目的：本研究旨在通过网络药理学与实验验证方法，探讨尿毒清颗粒治疗糖尿病肾病（Diabetic nephropathy, DN）所致足细胞损伤（podocyte injury）的药理学机制。
研究方法：从公共数据库中获取尿毒清颗粒的活性成分及其相关靶点，以及足细胞损伤、蛋白尿（proteinuria）与糖尿病肾病的相关基因。通过基因本体论（Gene Ontology, GO）、京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）以及蛋白质-蛋白质相互作用（Protein-Protein Interaction, PPI）网络分析，探究其潜在作用机制。采用高糖（High Glucose, HG）诱导的MPC5细胞损伤模型，给予尿毒清颗粒核心活性成分干预，以验证预测的靶点及信号通路。
研究结果：通过交集尿毒清颗粒靶点与疾病相关靶点，最终共筛选得到16个潜在治疗靶点；经PPI网络分析，其中7个被鉴定为核心靶点。KEGG富集分析结果显示，晚期糖基化终末产物（Advanced Glycation End Products, AGE）-晚期糖基化终末产物受体（Receptor for Advanced Glycation End Products, RAGE）信号通路为最为关键的信号通路。体外实验结果表明，给予尿毒清颗粒活性成分干预后，可显著保护MPC5细胞免受高糖诱导的细胞凋亡。此外，尿毒清颗粒可显著抑制高糖诱导的AGE-RAGE信号通路激活，并下调高糖处理的MPC5细胞中血管内皮生长因子A（Vascular Endothelial Growth Factor A, VEGF-A）、细胞间黏附分子1（Intercellular Adhesion Molecule 1, ICAM-1）、前列腺素内过氧化物合酶2（Prostaglandin-Endoperoxide Synthase 2, PTGS-2）及血管紧张素转换酶（Angiotensin-Converting Enzyme, ACE）的过度表达。
研究结论：尿毒清颗粒可能通过调控AGE/RAGE信号通路活性及多个基因的表达，减轻糖尿病肾病中的足细胞损伤。未来需开展进一步的临床与动物实验研究，以验证本研究结果。