Study on Qihuang Gushen Tongluo Prescription in Treating Diabetic Nephropathy Based on Bioinformatics and Animal Experiment
Objective:In this study,the mechanism of Qiguangushen Tongluo Prescription in the treatment of diabetic nephropathy(DKD)was investigated.Methods:The active components and targets of Qighuanggu Shen Tongluo Prescription and the DKD-re-lated targets were searched from GeneCards,TTD,DisGeNET,DrugBank,and other databases.The drug target points were mapped to the lesion target points,and the obtained intersection target points were constructed into a protein-protein interaction(PPI)net-work and active component-joint target network.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)en-richment analyses were performed.Animal experiments were conducted to verify the first three targets with more enrichment nodes in the core targets of DKD treated by Qihuanggushen Tongluo Prescription in the network pharmacology results.C57BL/6J mice were selected as the study objects,and streptozotocin(STZ)was injected intraperitoneally twice to prepare the animal model.After eight weeks,24 h urinary protein of the mice was measured,and the mice were divided into the normal group,model group,and model+Qihuanggushen Tongluo Prescription group.After eight weeks of intervention,insulin,C-peptide,urea nitrogen,and 24 h u-rinary protein were measured.The expressions of CTNNB1,JUN,and NCOA1 in mouse kidneys were detected by quantitative poly-merase chain reaction(qPCR)and Western blot.Results:GO and KEGG enrichment analyses showed that the effective targets of Qihuanggushen Tongluo Prescription in the treatment of DKD mainly involved the AGE-RAGE signaling pathway,lipid and athero-sclerosis,fluid shear stress and atherosis,neuroactive ligand-receptor interaction,etc.Animal experiments showed that Qihuanggush-en Tongluo Prescription could reduce the mRNA and protein expressions of CTNNB1,JUN,and NCOA1 in the kidney tissue of DKD mice and improve kidney injury.Conclusion:Through network pharmacology and experimental verification,it is found that Qi-huanggushen Tongluo Prescription may intervene in AGE-RAGE and other pathways to treat DKD by reducing the expression levels of CTNNB1,JUN,and NCOA1 proteins.