Mechanic evaluation of Jisheng Shenqi Wan (济生肾气丸) on calcium oxalate kidney stones: an integrated network pharmacology and metabolomics

doi:10.19852/j.cnki.jtcm.20250929.001

Abstract

Abstract:

OBJECTIVE: To understand the efficacy of Jisheng Shenqi Wan (JSSQW, 济生肾气丸) in treating calcium oxalate kidney stones (KS) and to investigate the mechanism of JSSQW action by combining network pharmacology with metabolomics analysis based on ultra-high performance liquid chromatography combined with tandem electrostatic field orbital trap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS).

METHODS: The chemical components of JSSQW absorbed into rat blood were identified by UHPLC-Q/Orbitrap HRMS. The identified components were introduced into the Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine platform to screen for target genes, followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and disease enrichment analysis. A KS rat model was generated using the oxalic acid precursor method to examine the efficacy of JSSQW for treating KS. Serum metabolomics was used to monitor changes in endogenous substances in KS rats after JSSQW intervention.

RESULTS: Twenty-three chemicals from JSSQW were identified in the blood of JSSQW gavage-administered rats. KEGG enrichment analysis predicted the top 20 signaling pathways affected by these 23 chemicals. Disease enrichment analysis showed that the target genes of these 23 chemicals were enriched in diseases of the urinary system and endocrine system, including kidney stones. In a KS rat model, JSSQW inhibited the aggregation of calcium oxalate crystals, reduced renal tubular injury, lowered the renal index, and improved biochemical indicators (blood creatinine, blood urea nitrogen). Serum metabolomics identified 25 differential metabolites that responded to JSSQW treatment. They were mainly lipids, with phosphatidylethanolamine and phosphorylcholine and their derivatives accounting for the highest proportion. Metabolic pathway analysis showed that the changes in differential metabolites were related to multiple metabolic pathways, especially sphingolipid metabolism and sphingolipid signaling pathways.

CONCLUSIONS: JSSQW can inhibit the aggregation of calcium oxalate crystals in the kidneys, reduce tubular injury, and improve kidney function in KS rats. Its mechanism of action may be related to regulating disordered metabolites and metabolic pathways, especially glycerol phospholipid metabolism, sphingolipid metabolism, and sphingolipid signaling.

Key words: kidney stones, network pharmacology, metabolomics, ultra-high performance liquid chromatography combined with tandem electrostatic field orbital trap high-resolution mass spectrometry, Jisheng Shenqi Wan

SHI Bing, LI Yang, JIANG Zhuocheng, QIN Guozheng, ZHAO Fan. Mechanic evaluation of Jisheng Shenqi Wan (济生肾气丸) on calcium oxalate kidney stones: an integrated network pharmacology and metabolomics[J]. Journal of Traditional Chinese Medicine, 2026, 46(2): 371-381.

Figures/Tables 4

Figure 1 UHPLC-Q/Orbitrap HRMS identification of JSSQW chemical components and network pharmacology analysis A: base peak chromatogram of the JSSQW serum group in positive mode; B: base peak chromatogram of the JSSQW Serum group in negative mode; C: KEGG pathway analysis; D: TTD and OMIM disease enrichment analysis. UHPLC-Q/Orbitrap HRMS: ultra-high performance liquid chromatography combined with tandem electrostatic field orbital trap high-resolution mass spectrometry; JSSQW: Jisheng Shenqi Wan; KEGG: kyoto encyclopedia of genes and genomes; TTD: Therapeutic Target Database; OMIM: Online Mendelian Inheritance in Man.

Figure 2 JSSQW inhibits calcium oxalate crystal deposition and renal tubular structural damage in KS rat kidneys and improves renal function A: Von Kossa staining of kidney sections from the different groups; B: appearance of kidneys in the different groups; C: kidney index in rats of different groups; D: serum creatinine levels in rats of different groups; E: blood urea nitrogen levels in rats of different groups; F: HE staining of kidney sections from the different groups. Black arrows indicate damaged renal tubules containing transparent deposits of calcium oxalate crystals; A1, B1, F1: Normal group; A2, B2, F2: Model group; A3, B3, F3: low-dose JSSQW group; A4, B4, F4: middle-dose JSSQW group; A5, B5, F5: high-dose JSSQW group; A6, B6, F6: K3cit group. Normal group: without calcium oxalate kidney stone modeling drug; Model group: without any treatment; low-dose JSSQW group: treated with 1.2 g/kg JSSQW for 4 weeks; middle-dose JSSQW group: treated with 2.4 g/kg JSSQW for 4 weeks; high-dose JSSQW group: treated with 4.8 g/kg JSSQW for 4 weeks; K3cit group: treated with 8 mL/kg K3cit for 4 weeks; JSSQW: Jisheng Shenqi Wan; KS: kidney stones; HE: hematoxylin and eosin staining. One-way analysis of variance was used to compare more than two groups, followed by the least significant difference test to detect differences between groups. The data are presented as the mean ± standard deviation (n = 8). Compared with Normal group, a P < 0.05; compared with Model group, b P < 0.05; compared with high-dose JSSQW group, c P < 0.05.

Figure 3 OPLS-DA and volcano plot of serum metabolomics analysis A: OPLS-DA scores of Model vs Normal group; B: OPLS-DA scores of H-JSSQW vs Model group; C: OPLS-DA permutation test analysis of Model vs Normal group; D: OPLS-DA permutation test analysis of H-JSSQW vs Model group; E: volcano plot of Model vs Normal group; F: volcano plot of H-JSSQW vs Model group. H-JSSQW: high-dose JSSQW group; Normal group: without calcium oxalate kidney stone modeling drug; Model group: without any treatment; high-dose JSSQW group: treated with 4.8 g/kg JSSQW for 4 weeks. JSSQW: Jisheng Shenqi Wan; KS: kidney stones; OPLS-DA: orthogonal partial least-squares discriminant analysis.

Figure 4 Classification and cluster analysis of differential metabolites A: venn diagrams of Model vs Normal group and H-JSSQW vs Model group; B: pie chart of classification of 25 differential metabolites; C: cluster analysis heatmap of 25 differential metabolites. JSSQW: Jisheng Shenqi Wan; H-JSSQW: high-dose JSSQW group; Normal group: without calcium oxalate kidney stone modeling drug; Model group: without any treatment; high-dose JSSQW group: treated with 4.8 g/kg JSSQW for 4 weeks.

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