Investigation on the antipyretic mechanism of Chaiqin Qingning capsule for the treatment of fever based on network pharmacology, molecular docking, and in vitro experimental validation.

Chaiqin Qingning Capsule (CQQNC), a traditional Chinese patent medicine, can effectively shorten the duration of fever and significantly improve fever symptoms. However, the mechanism of its antipyretic effect needs to be further elucidated. Therefore, we aimed to investigate the molecular mechanism of CQQNC in the treatment of fever. We used the network pharmacology method to analyze the mechanism of action of CQQNC in the treatment of fever and validated our study primarily by molecular docking. Finally, the predictive results were verified by IL‐1β‐induced bEnd.3 cells. The results showed that quercetin, kaempferol, cubebin, chenodeoxycholic acid, isorhamnetin, bilirubin, cholic acid, and baicalin were the major components of CQQNC against fever. A total of 381 common targets have been crossed by CQQNC for the treatment of fever. Furthermore, we found that CQQNC targets several deregulated genes in fever such as AKT1, COX2, AVP, cAMP, IL6, IL1B, TNF, mPGES1, and PI3K, biological functions such as endopeptidase, cytokine receptor binding, and phosphatase activity, and signaling pathways such as the PI3K‐Akt pathway and the AGE‐RAGE signaling pathway in diabetic complications. The docking study revealed that the core components of CQQNC both had high affinity for hub targets, especially the targets of COX‐2, cAMP, mPGES1, and PI3K proteins. To further investigate the mechanism of CQQNC, an Elisa assay and Western blot detection were performed as part of an in vitro study. Elisa's result showed that CQQNC can significantly decrease the expression levels of cPLA2, sPLA2, PGE2, cAMP, and 15‐PGDH after stimulating IL‐1β to bEnd.3 cells in a dose‐dependent manner (p <.01, p <.001, p <.0001). In addition, detection of the PGE2/ COX /cMAP pathway via immunoblotting showed that CQQNC can significantly downregulate the protein expression of COX‐1, COX‐2, EP3, cAMP, and mPGES1 in bEnd.3 (p <.0001, p <.001, p <.01). In conclusion, our study confirmed that the antipyretic mechanism of CQQNC affects the synthesis and secretion processes of PGE2 via the PGE2/ COX/cAMP pathway, providing insight into the antipyretic mechanism of CQQNC in clinical application. [ABSTRACT FROM AUTHOR]