[Molecular targets of Shenyan Fangshuai Liquid in treating diabetic kidney disease based on transcriptomics]

This study investigates the molecular targets of Shenyan Fangshuai Liquid(SYFSL) in the treatment of diabetic kidney disease(DKD) based on renal tissue transcriptomics. A DKD model was established in C57BL/6J mice by high-fat diet combined with streptozotocin induction. Mice were divided into six groups: normal group, model group, low-, medium-, and high-dose SYFSL groups, and dapagliflozin group, with 12 mice in each group. The low-,medium-, and high-dose SYFSL groups received oral gavage of the herbal suspension at corresponding doses. The dapagliflozin group received dapagliflozin suspension by gavage. The normal and model groups received an equal volume of physiological saline by gavage for 12 consecutive weeks. After treatment, blood glucose, kidney function indicators, and urine albumin-to-creatinine ratio(UACR) were measured. Kidney tissue pathological changes were observed, and RNA-Seq sequencing was used to analyze the renal transcriptome and identify differentially expressed genes(DEGs), followed by Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analyses. Additionally, weighted gene co-expression network analysis(WGCNA) was performed to identify module genes, and the intersection of DEGs and module genes was used to determine hub genes. Hub gene expression was validated by RT-qPCR and further confirmed with the NephroSeq database. Single-cell sequencing data were analyzed to explore hub gene expression in different cell types, and a hub gene-miRNA regulatory network was constructed. SYFSL significantly reduced blood glucose, kidney index, serum creatinine, blood urea nitrogen, and UACR in DKD mice, and improved kidney pathological damage. Transcriptome sequencing identified 1 284 DEGs, among which 95 genes were related to SYFSL intervention in DKD. Six hub genes(S100A6, EPHA2, TNC, SPON1, CLDN4, and TACSTD2) were identified. These genes were mainly enriched in biological processes such as extracellular matrix, basement membrane, and epithelial formation, as well as signaling pathways including extracellular matrix-receptor interaction, and showed correlations with kidney function and UACR. The expression patterns of hub genes in the NephroSeq database were consistent with sequencing results. Single-cell sequencing analysis revealed specific expression patterns of these hub genes in different cell types, and the constructed gene-miRNA network revealed complex interactions among them. This study, based on transcriptomic technology, demonstrates that SYFSL exerts therapeutic effects on DKD by regulating the expression of S100A6, EPHA2, TNC, SPON1, CLDN4, and TACSTD2 and their related signaling pathways, providing a scientific basis for its clinical use in DKD treatment.