Qingdi mixture alleviates radiation enteritis via the PI3K/AKT signaling pathway: An integrated study combining network pharmacology and experimental validation

Radiation enteritis (RE) is a common complication of radiotherapy for pelvic tumors, and Qindi Mixture (QDM) has been routinely used in tumor radiotherapy at the Affiliated Hospital of Qingdao University for the treatment of RE and alleviation of radiotherapy complications. However, the molecular mechanisms of QDM remain to be fully elucidated. Firstly, potential molecular targets of RE and QDM-related targets were systematically retrieved from established public databases such as TCMSP, using the results of mass spectrometry analysis from previous studies of QDM. Secondly, through comprehensive analysis of protein–protein interaction (PPI) network as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, we constructed target-pathway networks to identify potential therapeutic targets and associated signaling pathways. Subsequently, molecular docking was applied to validate the interactions between the QDM components and hub targets. To investigate the therapeutic potential, we established both in vitro RILI cell models and in vivo murine models. We elucidated molecular mechanisms of QDM’s protective effects against RE. Firstly, 387 related targets on RE and QDM were accessed through PPI network based on our previous LC-MS analysis on QDM. The core targets of the PPI network were mainly closely related to inflammation and transcriptional regulation. Secondly, GO and KEGG enrichment analyzes indicated that the pathways in cancer, PI3K-AKT signaling pathway, and Lipid and atherosclerosis might play key roles in the treatment of RE by QDM. Meanwhile, molecular docking showed that the core active components of QDM had stable binding to the main targets of RE. In vitro experiments showed that QDM inhibited the proliferation and migration of rectal cancer cells after radiotherapy. In vitro experiments demonstrated that QDM effectively inhibits the proliferation and migration of colorectal cancer cells after radiotherapy. In vivo, the downregulation of TP53 by QDM was associated with significant amelioration of RE, evidenced by an approximately 16% reduction in the AOD histopathological score ( p < 0.05). Western blot analysis showed that QDM treatment significantly reduced the activation ratios of p-PI3K/PI3K and p-AKT/AKT compared to the radiation group ( p < 0.05). The network pharmacology framework integrating computational prediction and experimental validation provides a novel approach for exploring the mechanism of QDM in treating RE.