Investigation of the mechanism of urolithin A in the treatment of renal fibrosis based on network pharma-cology and experimental verification
OBJECTIVE To investigate the efficacy and potential mechanism of urolithin A(UA)in the treatment of renal fibrosis induced by unilateral ureteral obstruction(UUO)based on network pharmacology,molecular docking and animal experi-ment.METHODS Network pharmacology techniques were used to analyze the mechanism of improving renal fibrosis action of UA.Auto Dock Tool software was used to perform molecular docking of UA with key targets to improve renal fibrosis.UUO was used to establish a rat renal fibrosis model.The regulatory effect of UA on renal fibrosis marker proteins was verified by immunohistochemical staining of fibronectin and collagen Ⅰ.The regulatory effects of UA on p-Akt1 and Akt1 were examined by immunofluorescence staining and Western blot.The effects of UA on the expression of PI3K and p-PI3K upstream,and GSK3βand p-GSK3βdownstream of Akt1 were examined by Western blot.RESULTS A total of 21 core targets were obtained by screening through network pharmacology techniques.Through PPI network topology analysis,GO biological function annotation and KEGG pathway enrichment analysis,it was found that UA could play a role in the treatment of renal fibrosis through signal pathways such as PI3K-Akt,p53 and key core targets such as Akt1,CCND1,CDK4,EGFR,CDK2,ERBB2,CDK6,GSK3B,BCL2L1,PTK2,ESR1 and PTGS2.Akt1 was the key target with the highest node degree in the PPI network study.Molecular docking further verified the high docking affinity of UA and Akt 1.Animal experiments showed that UA could signifi-cantly reduce the expression of fibronectin and collagen Ⅰ in UUO rats(P＜0.01),and inhibit the expression of p-Akt1(P＜0.01),as well as inhibited the phosphorylation of PI3K upstream and GSK3βdownstream of Akt1(P＜00.01).CONCLUSION UA can effectively alleviate renal fibrosis by inhibiting PI3K/Akt1/GSK3β signal pathway,and effectively reduce the expression of fibronectin and collagen Ⅰ in rat kidney,which provide scientific basis for further study of UA in the treatment of renal fibrosis.
urolithin Arenal fibrosisnetwork pharmacologymolecular dockingmechanism of actionPI3K/Akt1/GSK3β
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