TCM compatibility-driven mechanism of Shenshuaining (SSN) capsules against renal fibrosis: Integrated metabolomic and network analyses.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Shenshuaining (SSN), a formula documented in the Chinese Pharmacopoeia (2020), is used to treat chronic kidney disease (CKD) based on the TCM "Qi deficiency with turbid stasis" theory. Its composition of ten herbs follows the "Monarch, Minister, Assistant, and Guide" principle: Monarch drugs strengthen Qi and eliminate turbidity; Minister drugs resolve dampness and clear heat; Assistant drugs activate blood circulation; and the Guide drug harmonizes the formula. AIM OF THE STUDY: This study aims to elucidate the pharmacological mechanisms of SSN and its functional groups based on TCM compatibility principles, using integrated metabolomics and network pharmacology. MATERIALS AND METHODS: A Unilateral Ureteral Obstruction (UUO)-induced renal fibrosis rat model was used to evaluate SSN and the components of its four functional groups. Renal pathology as well as fibrosis, inflammation, and oxidative stress biomarkers were analyzed, and HPLC-MS was used to identify SSN's components, while network pharmacology predicted targets. Metabolomics revealed differential metabolites, and compound-target-metabolite networks were constructed, while molecular docking was used to assess interactions between key compounds and targets, supported by biochemical validation of functional group mechanisms. The protective effects and mechanisms of SSN and its functional groups were further validated in TGF-β-induced HK-2 cells. RESULTS: SSN significantly attenuated renal fibrosis, inflammation, and oxidative stress, as confirmed by histopathology and biomarker assays. Metabolomics revealed that the functional groups of SSN, Monarch, Minister, Assistant, and Guide, showed sequentially reduced regulatory effects on differentially metabolites in both serum and renal tissues, enriched pathways were primarily associated with lipid-related metabolism. Integrated analysis of renal fibrosis related targets associated with 63 compounds and 50 differential metabolites revealed that CA2, NOS3, and PLA2G2A serve as specific targets for the Monarch, Minister, and Guide groups respectively, while PPARG was identified as a common target regulated by the Monarch, Minister, and Assistant group to collaboratively suppressed lipid accumulation. Each functional group inhibited fibrosis via its respective targets, illustrating integrated synergistic actions and target-specific anti-fibrotic mechanisms. CONCLUSIONS: The functional groups of SSN ameliorated renal fibrosis through their respective targets, while also synergistically modulating lipid-related metabolism to attenuate fibrosis. This integrated multi-target mechanism systematically validates the TCM compatibility principle and provide a scientific rationale for the composition of SSN.