Study on the Potential Mechanism of Astragaloside IV on Renoprotection in Db/Db Mice via Network Pharmacology and Experimental Validation.

Abstract

BACKGROUND/AIMS: Diabetic nephropathy (DN) is a highly predominant and critical microvascular complication associated with diabetes mellitus. Astragaloside IV (AS-IV), the main active component in Radix Astragali, is associated with various pharmacological effects, including on DN. Nevertheless, the fundamental mechanism through which AS-IV ameliorates DN is still unclear. Therefore, this research seeks to explore the molecular mechanisms underpinning the therapeutic effectiveness of AS-IV for DN using network pharmacology and experimental validation. METHODS: Initially, DN mouse models were created and administered AS-IV treatment. Key metabolic parameters were assessed, and pathological alterations in the mice's kidneys were examined. Subsequently, the GEO database and other online public databases were utilized to detect DN-correlated targets and the primary AS-IV targets. A protein-protein interaction (PPI) network of overlapping targets was created to pinpoint core targets using Cytoscape software. These core targets underwent GO and KEGG analyses. Differential gene analysis and ROC curve analysis were then conducted on the core targets. Finally, western blotting was conducted to confirm the expression changes of core targets and pathway proteins. RESULTS: AS-IV treatment markedly reduced fasting blood glucose (FBG), blood urea nitrogen (BUN), body weight (BW), urinary albumin excretion (UAE), and serum creatinine (Scr) levels and alleviated histopathological alterations in the DN mice's kidneys. Twelve key core targets were identified through GEO data analysis and network pharmacology, with four core targets (MMP-9, MPO, IL-6, and IL-1β) emerging after more stringent screening. Enrichment analysis illustrated that these genes predominantly participated in biological processes, such as regulating oxidative stress and inflammatory responses, and were part of pathways like TNF, IL-17, and AGE-RAGE signaling in DM complications. Western blotting further displayed that AS-IV treatment downregulated the levels of DN-affected proteins MMP-9, MPO, IL-1β, IL-6, and TNF-α. CONCLUSIONS: Overall, combining the network pharmacology and experimental validation offers a precise elucidation of the molecular mechanism by which AS-IV treats DN. Additionally, it proposes a novel approach for identifying the active components of traditional Chinese medicine.