Chemical profiles and mechanisms insights into Liuwei Dihuang-based formulas in diabetic nephropathy treatment: An integrated transcriptomics with in vitro experiments

Abstract

Liuwei Dihuang formula(LWDH) is widely accepted as a remedy for tonifying kidney due to its medicine and food dual nature. LWDH and its various derivative prescriptions(JWDHs) with edible herbs were used in the clinical treatment of diabetic nephropathy(DN). Although, the mechanisms of LWDH being increasingly clarified, mechanisms of various JWDHs remained largely unexplored. The purpose of this study is to explore the mechanisms of Guifu Dihuang(GFDH), Qiju Dihuang(QJDH), and Zhibai Dihuang(ZBDH) formulas in treating DN. Transcriptomics, including GO, KEGG, and GSEA, was used to identify potential targets and pathways, RT-qPCR for further validation. UPLC-MS/MS was utilized to uncover chemical profiling of GFDH, QJDH and ZBDH. Molecular docking and molecular dynamics simulations were conducted to screen components and assess the stability of drug-target binding. As results, GFDH, QJDH and ZBDH exhibited multi-target and multi-pathway characteristics in treating DN. They collectively regulated the CDKN1A/FOXO signaling pathway, and QJDH uniquely modulated the PGK1/HIF-1 signaling pathway. UPLC-MS/MS identified 72 components across GFDH, QJDH and ZBDH, with 45 common and 7 unique to QJDH. Molecular docking indicated that unique components in QJDH, including neochlorogenic acid, chlorogenic acid, naringenin, kaempferide, and oroxylin A, are potential agents for intervening DN through PGK1. Molecular dynamics simulation confirmed that neochlorogenic acid exhibited stable binding with PGK1, further supporting the validation of the unique mechanism of action of QJDH. This study highlights the diverse mechanisms of GFDH, QJDH, and ZBDH in combating DN, offering a theoretical basis for clinical use and a foundation for developing food therapy approaches.