基于网络药理学和裸鼠体内实验探讨补脾胃泻阴火升阳汤通过HIF-1α调节糖酵解途径抗结肠癌的机制OA

Objective To investigate the mechanisms of Bupiwei Xieyinhuo Shengyang Decoction(BpwXyhSyD)in treating colon cancer through network pharmacology combined with in vivo experiments.Methods ① Using TCMSP,GeneCards,OMIM,and DrugBank databases to search for active ingredients and colon cancer-related targets of BpwXyhSyD,a protein-protein interaction(PPI)network was constructed using the STRING platform,while Gene Ontology(GO)and Kyoto Gene and Genome Database(KEGG)enrichment analysis were performed.②Sixty BALB/C nude mice were subcutaneously injected with human colon concer HCT-116 cells to establish a colon cancer model and randomly divided into four groups:model group,traditional Chinese medicine(TCM)group(20.93 g·kg-1),Western medicine group(capecitabine:390 mg·kg-1),and combined TCM-Western medicine group(TCM:20.93 g·kg-1;capecitabine:390 mg·kg-1).Mice in each group received corresponding drug interventions for a total of 2 weeks.Tumor growth was monitored by measuring tumor weights in each group.Histopathological changes in tumor tissues were observed using hematoxylin and eosin(HE)staining.Immunohistochemical techniques were employed to detect the expression of HIF-1α,GLUT1,HK2,and LDHA in tumor tissues.Western blot and RT-qPCR analyses were conducted to assess the protein and mRNA expression levels of HIF-1α,GLUT1,HK2,and LDHA in tumor tissues.Results ①A total of 141 intersection genes between the BpwXyhSyD and colon cancer were successfully identified.Among these,p53,AKT1,EGFR,and HIF-1α may represent key therapeutic targets for the BpwXyhSyD in colon cancer treatment.The underlying mechanisms primarily involve signaling pathways such as p53,VEGF and HIF-1.② Compared with the model group,HE staining and immunohistochemistry revealed varying degrees of cellular disruption and necrosis in tumor tissues across all drug intervention groups.Western blot analysis demonstrated significantly reduced relative expression levels of HIF-1α,GLUT1,HK2,and LDHA proteins in tumor tissues treated with each intervention.RT-qPCR results demonstrated significantly reduced mRNA expression levels of LDHA in both the Western medicine group and the TCM-Western medicine group(P<0.05),along with significantly reduced mRNA expression of HIF-1α,GLUT1,and HK2 in all drug intervention groups(P<0.05).Furthermore,compared with the model group and the TCM group,the tumor tissue in the TCM-Western medicine group exhibited more pronounced destruction,along with lower expression levels of HIF-1α,GLUT1,HK2,and LDHA proteins(P<0.05).Conclusion The use of BpwXyhSyD synergistically inhibits glycolysis,thereby suppressing the growth of colon cancer xenografts.The mechanism may involve downregulating HIF-1α expression and inhibiting the expression of key enzymes in glycolysis,including GLUT1,HK2,and LDHA.