Mechanism of Shenqi Pills(肾气丸)in Treatment of Ovulation Disorder with Shenxuzheng(肾虚证)Based on AGE-RAGE Signaling Pathways
Objective:To study the mechanism of Shenqi Pills(肾气丸)in improving ovulation disorder(OD)through network pharmacology and to validate the findings through animal experiments.Methods:Active components and related targets were screened by network pharmacolo-gy,and a"drug-active component-target"network and a protein-protein interaction(PPI)network was constructed.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed on the intersecting targets using R language.According to the results of network pharmacology,animal experiments were conducted.A model was established by intramuscular injection of hydrocortisone in both hind limbs.After successful modeling,Shenqi Pills were administered for 14 days.Vaginal smears were used to observe estrous cy-cles.HE staining was used to observe histopathological changes in ovarian tissue.TUNEL staining was used to detect apoptosis in ovarian granulosa cells.Serum levels of FSH,LH,E2,P,IL-1β,IL-6,and TNF-α were measured to assess hormone levels and inflammatory response.Western blot was used to detect the expression of AGEs,RAGE,ERK1/2,and C-JUN proteins in ovarian tissue.Results:Shenqi Pills were found to contain 117 active components and 93 therapeutic targets for treating OD.Quercetin,diosgenin,β-sitosterol,(+)-epicatechin,and di-hydroquercetin were identified as potential key components for OD intervention.The PPI network identified 20 core targets.GO enrichment a-nalysis showed the top three molecular functions were DNA-binding transcription factor binding,RNA polymerase Ⅱ-specific DNA-binding transcription factor binding,and DNA-binding transcription activator activity.The top three biological processes were response to external biot-ic stimulus,epithelial cell proliferation,and oxidative stress response.The top three cellular components were membrane raft,membrane mi-crodomain,and vesicle lumen.KEGG analysis indicated that the AGE-RAGE signaling pathway played a crucial role in the treatment of OD with Shenqi Pills.Animal experiments demonstrated that intervention with Shenqi Pills improved estrous cycle disorders in rats,increased the number of follicles at various stages,reduced follicular atresia,inhibited apoptosis of ovarian granulosa cells,normalized levels of FSH,LH,E2,and P hormones,reduced inflammatory responses,and downregulated the expression of AGEs,R AGE,ERK1/2,and C-JUN proteins,there-by regulating the AGE-RAGE signaling pathway to improve OD.Conclusion:Shenqi Pills may improve OD by regulating the expression of AGEs,RAGE,ERK1/2,and C-JUN proteins in the AGE-RAGE signaling pathway,thereby improving hormone levels,reducing inflammation,and inhibiting granulosa cell apoptosis.
Shenqi Pills(肾气丸)Network PharmacologyShenxuzheng(肾虚证)Ovulatory DisorderAdvanced Glycation End Prod-ucts/Glycation End Products Receptor Signaling Pathway
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