Protective Effects and Mechanisms of CHEBULAE FRUCTUS Water Extract in Rats with Hyperuricemic Nephropathy
Objective:To investigate the effects and preliminary mechanisms of CHEBULAE FRUCTUS water extract in improving hyperuricemic ne-phropathy in rats.Methods:Active components and target information of CHEBULAE FRUCTUS were obtained through the Traditional Chi-nese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP),and targets related to hyperuricemic nephropathy were col-lected from the GeneCards database.Subsequently,protein-protein interaction (PPI) analysis was performed by using the STRING database.Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed via Metascape platform."CHEBULAE FRUCTUS-active components-intersecting targets-disease" network diagram was constructed by Cytoscape 3.7.1 soft-ware.Autodock Vina software was employed for the molecular docking of core components of CHEBULAE FRUCTUS with key targets,and the ADMET properties and drug-likeness of the core components were predicted by the pkCSM database.A rat model of hyperuricemic nephropa-thy was established by using potassium oxonate combined with 10％ fructose,and the protective effect of CHEBULAE FRUCTUS water extract was verified through renal function index tests of serum and urine,as well as HE,PAS,and Masson staining of renal tissue.The effects of CHEBULAE FRUCTUS water extract and its core components on the uric acid-induced HK-2 renal injury cell model and key targets were vali-dated using methods such as CCK8,RT-PCR,and Western blot.Results:Seven active components of CHEBULAE FRUCTUS were identified,with 70 component targets,878 hyperuricemic nephropathy-related targets,and 22 drug-disease intersecting targets.GO enrichment analysis indicated that CHEBULAE FRUCTUS mainly affected biological processes such as leukocyte apoptosis,cellular responses to organic cyclic compounds,and responses to tumor necrosis factors.It exerted molecular functions such as binding to specific protein domains,cysteine-type endopeptidase activity in the apoptotic signaling pathway,and BH3 domain binding,involving cellular components such as membrane mi-crodomains,transcription regulation complexes,and receptor complexes and thereby playing a role in treating hyperuricemic nephropathy.KEGG enrichment analysis revealed that the main pathways involved in the treatment of hyperuricemic nephropathy by CHEBULAE FRUCTUS were the P53 signaling pathway,apoptosis signaling pathway,etc.TP53,ESR1,CASP8,CASP9,and CASP3 were key targets in the treatment of hyperuricemic nephropathy by CHEBULAE FRUCTUS.Additionally,molecular docking results showed that the core components of CHE-BULAE FRUCTUS had good binding ability with key targets,and ADMET properties and drug-likeness analyses indicated that all core compo-nents had good drug properties.Animal experiments indicated that CHEBULAE FRUCTUS water extract significantly reduced the contents of serum uric acid (SUA),serum creatinine (Scr),blood urea nitrogen (BUN),and 24-hour urinary protein levels in rats with hyperuricemic nephropathy (P<0.05 or P<0.01) and improved pathological damage of renal tissue.Cell experiments showed that CHEBULAE FRUCTUS water extract and its core components effectively enhanced the viability of uric acid-induced HK-2 cells and improved cell condition.At the molecular level,CHEBULAE FRUCTUS water extract significantly down-regulated the gene expression of Casp8 and Casp3 (P<0.01),and the cheilanthifoline significantly down-regulated Tp53 gene expression (P<0.01).Quinidine sulfate significantly down-regulated the gene and protein expression of Tp53,Esr1,Casp8,Casp9,and Casp3 (P<0.05 or P<0.01).Conclusion:CHEBULAE FRUCTUS water extract can im-prove renal injury in rats with hyperuricemic nephropathy,and its mechanism of action may be based on its core components Quinidine sulfate acting through the anti-apoptotic pathway.
CHEBULAE FRUCTUS water extractHyperuricemic nephropathyNetwork pharmacologyMolecular dockingApoptosisQuinidine sulfate
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