基于网络药理学、分子对接技术和动物实验探讨小柴胡汤对氟中毒所致肾损伤的保护作用OA

Objective:To explore the protective mechanisms of Xiao Chaihu Decoction against fluoride-induced renal injury using network pharmacology,molecular docking,and animal experiments.Methods:Thirty-two male Sprague-Dawley(SD)rats were randomly divided into four groups(n=8 per group):normal control(NC)group,sodium fluoride(NaF)group,Xiao Chaihu Decoction intervention(XCHT+NaF)group,and Xiao Chaihu Decoction negative control(XCHT)group.The rats were allowed to freely drink NaF solution at a concentration of 100 mg/L to establish a chronic fluorosis rat model.The effect of Xiao Chaihu Decoction on renal function,oxidative stress levels and pathological changes in kidney tis-sue were observed.Active compounds and potential targets of XCHT were retrieved from the TCMSP database,ChemSpider,and SwissTargetPrediction.Targets associated with fluoride poisoning and kidney injury were obtained from the GeneCards database.Intersection targets were identified,and a protein-protein interaction(PPI)network was constructed using the STRING database and visualized with Cytoscape 3.10.2 to identify core targets.Gene Ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were performed via the DAVID database.A"XCHT-active compound-core target-pathway"network was constructed using Cytoscape.Molecular docking and visualiza-tion of the main active compounds of XCHT and key targets were performed using software such as AutoDock Tools 1.5.7 and Chimera 1.17.3.Results:(1)Animal experiments:Compared with the NaF group,the XCHT+NaF group showed signifi-cantly reduced serum uric acid(UA)and creatinine(Scr)levels,decreased malondialdehyde(MDA)content,and super-oxide dismutase(SOD)activity in renal tissue(P<0.05 or P<0.01),suggesting that XCHT alleviates fluoride-induced renal injury and oxidative stress.Compared with NaF group,the range of renal lesions and the degree of inflammatory cell infiltra-tion were significantly improved in the XCHT+NaF group.(2)Network pharmacology analysis:A total of 679 XCHT-related targets,715 fluoride poisoning-related targets,and 8,417 kidney injury-related targets were identified,yielding 203 inter-secting targets.Quercetin,kaempferol,stigmasterol,STAT3(signal transducer and activator of transcription 3),JUN(activa-tor protein-1),and TP53(tumor protein p53)were identified as core active compounds,GO enrichment analysis indicated that biological processes(BP)were primarily involved in positive regulation of cAMP-dependent protein kinase activity,negative regulation of cholesterol storage,and inhibitory G-protein-coupled serotonin receptor signaling.Cellular compo-nents(CC)were mainly associated with heterochromatin and cyclin-dependent protein kinase holoenzyme complexes.Mo-lecular functions(MF)included caffeine oxidase activity,dopamine receptor binding,and estrogen response element bind-ing.KEGG pathway analysis highlighted 20 significantly enriched pathways,including the IL-17 signaling pathway,PI3K-Akt signaling pathway,and HIF-1 signaling pathway.Molecular docking results showed that all three core compounds exhib-ited favorable binding affinities with JUN,STAT3,and TP53.Specifically,quercetin exhibits a binding energy of-6.68 kcal/mol with JUN;Kaempferol shows a binding energy of-6.54 kcal/mol with TP53;and stigmasterol demonstrates a binding en-ergy of-10.68 kcal/mol with STAT3.Conclusion:Xiao Chaihu Decoction may exert protective effects against fluoride-induced renal injury through multi-target and multi-pathway mechanisms.