Effect and Mechanisms of Huangqi-Shanzhuyu in the Treatment of Diabetic Nephropathy based on Network Pharmacology and In Vitro Experiments

Abstract

Background: Huangqi-Shanzhuyu (HS), a classic combination of Chinese herbal formulae, has been widely used for the treatment of diabetic nephropathy (DN). However, its pharmacological mechanism of action is still unclear. background: Huangqi-Shanzhuyu (HS) is a classic combination in Chinese herbal formulas, which has been widely used in the treatment for diabetic nephropathy (DN). However, its pharmacological mechanism is still unclear. Methods: The active ingredients of HS and their potential targets were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the DN-related targets were determined from GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGkb, and Therapeutic Target Database (TTD). The Cytoscape software was used to construct a herb-disease-target network and screen core genes. STRING was employed to generate a protein-protein interaction (PPI) network. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to predict the mechanism of action of HS in DN. Animal experiments and molecular docking were used to verify the potential mechanism. objective: To obtain the active ingredients and targets of HS and the targets of DN. To construct the herb-disease-target network, protein interaction network and screen core genes. To predict the mechanism of HS treatment on DN. To verify the potential mechanism by animal experiment and molecular docking. Results: In total, 40 active ingredients and 180 effective targets of HS in DN were identified and 1115 DN-related targets were retrieved. From the PPI network, VEGFA, AKT1, IL6, IL1B, TP53, MMP9, PTGS2, CASP3, EGF and EGFR were identified as core genes. The anti-DN mechanism mainly involved multiple signaling pathways such as AGEs-RAGE. Animal experiments and molecular docking analysis confirmed that HS downregulated the expression of IL-1 and IL-6 via kaempferol-mediated inhibition of JNK1 phosphorylation. method: The active ingredients and targets of HS were obtained through TCMSP. The targets of DN were obtained by GeneCards, OMIM, PharmGkb, and TTD databases. Cytoscape software was used to construct the herb-disease-target network and screen core genes. STRING was used to construct protein interaction network. GO and KEGG enrichment analysis were applied to predict the mechanism of HS treatment on DN. Animal experiment and molecular docking were used to verify the potential mechanism. Conclusions: HS exhibits a therapeutic effect in DN through its multiple ingredients that act on several targets and multiple signaling pathways, including AGEs-RAGE. result: There were 40 active ingredients and 180 effective targets of HS. A total of 1115 DN-related targets were retrieved. From the PPI network, VEGFA, AKT1, IL6, IL1B, TP53, MMP9, PTGS2, CASP3, EGF, and EGFR were identified as corn genes. The mechanism of anti-DN mainly involved multiple signaling pathways such as AGEs-RAGE. Animal experiments and molecular docking analysis confirmed that HS reduced the expressions of IL-1 and IL-6, which was related to the inhibition of JNK1 phosphorylation by kaempferol. conclusion: HS has the therapeutic on anti-diabetic nephropathy by multiple ingredients, multiple targets and multiple signaling pathways including AGEs-RAGE. other: None.