基于网络药理学、分子对接和实验验证揭示白藜芦醇治疗糖尿病肾病的机制。

BACKGROUND: Diabetic kidney disease (DKD) has been the leading cause of chronic kidney disease in developed countries. Evidence of the benefits of resveratrol (RES) for the treatment of DKD is accumulating. However, comprehensive therapeutic targets and underlying mechanisms through which RES exerts its effects against DKD are limited. METHODS: Drug targets of RES were obtained from Drugbank and SwissTargetPrediction Databases. Disease targets of DKD were obtained from DisGeNET, Genecards, and Therapeutic Target Database. Therapeutic targets for RES against DKD were identified by intersecting the drug targets and disease targets. GO functional enrichment analysis, KEGG pathway analysis, and disease association analysis were performed using the DAVID database and visualized by Cytoscape software. Molecular docking validation of the binding capacity between RES and targets was performed by UCSF Chimera software and SwissDock webserver. The high glucose (HG)-induced podocyte injury model, RT-qPCR, and western blot were used to verify the reliability of the effects of RES on target proteins. RESULTS: After the intersection of the 86 drug targets and 566 disease targets, 25 therapeutic targets for RES against DKD were obtained. And the target proteins were classified into 6 functional categories. A total of 11 cellular components terms and 27 diseases, and the top 20 enriched biological processes, molecular functions, and KEGG pathways potentially involved in the RES action against DKD were recorded. Molecular docking studies showed that RES had a strong binding affinity toward PPARA, ESR1, SLC2A1, SHBG, AR, AKR1B1, PPARG, IGF1R, RELA, PIK3CA, MMP9, AKT1, INSR, MMP2, TTR, and CYP2C9 domains. The HG-induced podocyte injury model was successfully constructed and validated by RT-qPCR and western blot. RES treatment was able to reverse the abnormal gene expression of PPARA, SHBG, AKR1B1, PPARG, IGF1R, MMP9, AKT1, and INSR. CONCLUSIONS: RES may target PPARA, SHBG, AKR1B1, PPARG, IGF1R, MMP9, AKT1, and INSR domains to act as a therapeutic agent for DKD. These findings comprehensively reveal the potential therapeutic targets for RES against DKD and provide theoretical bases for the clinical application of RES in the treatment of DKD.

背景：糖尿病肾病（DKD）已成为发达国家慢性肾脏病的主要病因。越来越多的证据表明白藜芦醇（RES）对 DKD 的治疗有益。然而，综合的治疗靶点以及 RES 发挥作用的潜在机制仍有限。

方法：从 Drugbank 和 SwissTargetPrediction 数据库中获取 RES 的药物靶点。从 DisGeNET、Genecards 和 Therapeutic Target Database 中获取 DKD 的疾病靶点。通过 intersecting 药物靶点和疾病靶点来鉴定 RES 治疗 DKD 的治疗靶点。使用 DAVID 数据库进行 GO 功能富集分析、KEGG 通路分析和疾病关联分析，并通过 Cytoscape 软件可视化。使用 UCSF Chimera 软件和 SwissDock webserver 对 RES 与靶标之间的结合能力进行分子对接验证。采用高糖（HG）诱导的足细胞损伤模型、RT-qPCR 和 Western blot 验证 RES 对靶蛋白作用的可靠性。

结果：在 86 个药物靶点和 566 个疾病靶点的交集后，获得了 25 个 RES 治疗 DKD 的治疗靶点。并且将靶蛋白分为 6 个功能类别。共记录了 11 个细胞成分术语和 27 种疾病，以及 20 个最富集的生物过程、分子功能和 KEGG 通路，这些通路可能涉及 RES 对 DKD 的作用。分子对接研究表明 RES 对白藜芦醇受体（PPARA）、雌激素受体 1（ESR1）、溶质载体家族 2 成员 1（SLC2A1）、性激素结合球蛋白（SHBG）、雄激素受体（AR）、醛酮还原酶 1B1（AKR1B1）、过氧化物酶体增殖物激活受体 γ（PPARG）、胰岛素样生长因子 1 受体（IGF1R）、核因子 κB 亚基 p65（RELA）、磷酸肌醇 3 激酶催化亚基（PIK3CA）、基质金属蛋白酶 9（MMP9）、蛋白激酶 B（AKT1）、胰岛素受体（INSR）、转甲状腺素蛋白（TTR）和细胞色素 P450 2C9 结构域具有很强的结合亲和力。成功构建并验证了 HG 诱导的足细胞损伤模型，通过 RT-qPCR 和 Western blot 验证。RES 治疗能够逆转 PPARA、SHBG、AKR1B1、PPARG、IGF1R、MMP9、AKT1 和 INSR 的异常基因表达。

结论：RES 可能通过靶向 PPARA、SHBG、AKR1B1、PPARG、IGF1R、MMP9、AKT1 和 INSR 结构域来作为 DKD 的治疗药物。这些发现全面揭示了 RES 治疗 DKD 的潜在治疗靶点，并为 RES 在 DKD 治疗中的临床应用提供了理论依据。