Exploration of Molecular Mechanisms of Shenqi Fuzheng Injection in Improving Cisplatin-Induced Acute Kidney Injury by Combining Network Pharmacology and Transcriptomics
Abstract:Objective:Using network pharmacology and transcriptomics methods,this paper explores the potential targets of Shenqi Fuzheng Injection in improving cisplatin-induced acute kidney injury (AKI), providing experimental basis for further clinical guidance. Methods:①Network pharmacology analysis:The HERB Database (HERB), Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), Herbal Ingredients' Targets (HIT), Swiss Target Prediction, PubChem BioAssay Database (PubChem), UniProt Knowledgebase (UniProt), and Protein-protein Interaction (PPI) Database (STRING) were applied to obtain potential targets for the main components of Shenqi Fuzheng Injection. The Online Mendelian Inheritance in Man (OMIM), Human Gene Database (GeneCards), and Therapeutic Target Database (TTD) were used to obtain potential targets for AKI. Using STRING database and Cytoscape 3.9.1 software, a PPI network diagram for common potential targets of the main components of Shenqi Fuzheng Injection and AKI was constructed and network topology analysis was performed. By using R language software, the gene ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. ② Animal experiment transcriptomics analysis: The 4T1 tumor-bearing mouse model of breast cancer were established and divided into four groups (the model group, the Shenqi Fuzheng Injection group, the cisplatin group, the Shenqi Fuzheng Injection combined with cisplatin group). After four weeks of administration, blood was taken to detect creatinine (CRE) and urea nitrogen (BUN), and kidney tissue was taken for transcriptome sequencing analysis. Real time quantitative polymerase chain reaction (qRT-PCR) was used for experimental verification. Results: A total of 65 common potential targets of the main components of Shenqi Fuzheng Injection and AKI were predicted through a network pharmacology database and Cytoscape 3.9.1 software. The PPI network diagram and common potential target network diagram indicated that there are multiple functional relationships between the main components of Shenqi Fuzheng Injection and AKI with 65 common potential targets. The results of GO functional enrichment analysis and KEGG pathway enrichment analysis indicated that the improvement of AKI by Shenqi Fuzheng Injection is closely related to oxidative and antioxidant, inflammatory and immune responses, as well as T cell receptor signaling pathways and hypoxia inducible factor-1 signaling pathways. The CRE and BUN levels in serum in the cisplatin group were higher than those in the model group (P<0.05). The CRE level in the Shenqi Fuzheng Injection group was significantly lower than that in the model group (P<0.05). The levels of CRE,BUN in serum in the cisplatin group were higher than those in the model group (P<0.05). The CRE level in serum in the Shenqi Fuzheng Injection group was lower than that in the model group (P<0.05). The levels of CRE, BUN in serum in the Shenqi Fuzheng Injection group were lower than those in the cisplatin group (P<0.05). The levels of CRE,BUN in serum in the Shenqi Fuzheng Injection combined with cisplatin group were lower than those in the cisplatin group (P<0.05). The CRE level in serum in the Shenqi Fuzheng Injection combined with cisplatin group was higher than that in the Shenqi Fuzheng Injection group (P<0.05). There was no significant difference in the BUN levels in serum between the Shenqi Fuzheng Injection combined with cisplatin group and the Shenqi Fuzheng Injection group (P>0.05). The HE staining results of kidney tissue in the four groups of mice showed that Shenqi Fuzheng Injection improved cisplatin-induced kidney injury. Combining network pharmacology and transcriptomics analysis, nine common potential targets were identified, among which the down-regulated genes of Shenqi Fuzheng Injection included xanthine dehydrogenase, interleukin-10, E-cadherin, and interferon- γ, estrogen receptor α, prostaglandin endoperoxide synthase 1, and up-regulated genes included peroxisome proliferator-activated receptors- γ, interleukin-6, and CD4+ T cells. Conclusion: Shenqi Fuzheng Injection can improve cisplatin-induced AKI by affecting oxidative stress, inflammation, and immune response, and has the advantages of multiple pathways and targets.