Identifying active substances of Huangqin Decoction regulating intestinal epithelial barrier dysfunction by mass spectrometry networking and multiscale models

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Introduction: Huangqin decoction (HQD) is a traditional Chinese medicine prescription recorded in the ''Shang Han Lun'', a decoction composed of huangqin, chishao, gancao and dazao, which has been widely used to relieve symptoms of gastrointestinal diseases, such as ulcerative colitis (UC) and typhoid fever. However, its active components is yet to be clarified. Methods: The chemical composition of HQD was analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by network pharmacology prediction. A colonic cell line and mouse colonic organoid inflammation model were established to observe the effects of HQD and its components on inflammatory factor expression and intestinal barrier function. The efficacy of the aqueous fraction of HQD (HQD-far-A) was further validated in a zebrafish enteritis model by assessing intestinal oxidative stress levels, neutrophil aggregation levels, and goblet cell counts. Results: In the Caco-2 cell inflammation model, HQD and its individual components all reduced the pharmacological effects of elevated inflammatory factors. Treatment with HQD-fra-A significantly protected the intestinal epithelial barrier, including tight junction proteins ZO-1 and Occludin, and this effect was validated in a DSS-induced intestinal organoid inflammation model. In vitro experiments demonstrated that HQD-fra-A reduced neutrophil aggregation and intestinal oxidative stress caused by intestinal inflammation in zebrafish, mitigated intestinal injury, and protected goblet cells to maintain the intestinal barrier. Discussion: HQD has been validated as an effective treatment for colon inflammation by protecting the intestinal barrier. The multi-scale model for drug efficacy validation provides a solid foundation and new insights for advancing drug efficacy evaluations in future research.

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