Can "Fire" Change Medicinal Effects? Processing heat transforms the efficacy of Chaihu from antipyretic to hepatic-soothing

Traditional Chinese Medicine (TCM) from centuries past asserts that unprocessed Chaihu (Bupleurum chinense DC.) primarily exhibits exterior-releasing properties, hepatic soothing, and is advisable for use in cases of Yin deficiency and bone-steaming sensation. To validate this age-old clinical experience, we employed modern biomedical methodologies such as TCM network pharmacology and molecular docking. Through network pharmacology research, specific targets (AKT, IKKs, IκBα, CDK1, VEGKA) associated with Chaihu in treating diseases were identified within the PI3K-Akt signaling pathway. Initial conclusions were drawn through target network survival validation, network immunohistochemistry verification, and molecular docking studies, suggesting that processing Chaihu with heat alters its efficacy. Subsequent validation using PCR and WB experiments on unprocessed Chaihu, turtle blood-processed Chaihu, vinegar-processed Chaihu, and Quercetin against the selected PI3K-Akt signaling pathway further confirmed that processing Chaihu with heat enhances its inhibitory effect on the PI3K-Akt signaling pathway. This interference aids in transcriptional regulation of inflammatory factors, thereby augmenting the hepatic soothing effects, particularly enhancing the hepatic soothing and depression-relieving efficacy post-processing. Additionally, turtle blood-processed Chaihu and vinegar-processed Chaihu exhibited a more pronounced impact on AKT and IKKs compared to unprocessed Chaihu. Conclusions: This substantiates that processing Chaihu with heat weakens its exterior-releasing and antipyretic effects while strengthening its hepatic soothing and depression-relieving properties.