Dahuang Huanglian Decoction alleviates dysbiosis by inhibiting GBP5/NLRP3 signaling pathway-mediated pyroptosis of colonic epithelial cells.

Ethnopharmacological Relevance: Currently, the prevalence of dysbiosis is increasing, but its treatment options are limited. Dahuang Huanglian Xiexin Decoction (XXD) is a traditional herbal prescription recorded in the Treatise on Typhoid Fever, with a longstanding application in the treatment of digestive system diseases. It consists mainly of three classical chinese medicinal herbs: Dahuang (Rheum palmatum L.), Huangqin (Scutellaria baicalensis Georgi), and Huanglian (Coptis chinensis Franch.). Previous studies demonstrated the efficacy of XXD in treating dysbiosis. However, the exact underlying mechanism requires further investigation.

Aim Of The Study: The effects of XXD were evaluated in this study to determine its impact on dysbiosis and to reveal the potential mechanisms underlying its alleviation using proteomics and transcriptomics.

Materials And Methods: The components of XXD were identified through UPLC-Q-TOF-MS. Dysbiosis mice were established by mixing antibiotic solutions, and XXD was employed as the therapeutic agent in the intervention. Body weight changes, diarrhea rates, and histopathology were evaluated to determine the therapeutic effects of XXD. Proteomics and transcriptomics were subsequently employed to further elucidate the mechanisms underlying the therapeutic effects of XXD on dysbiosis. Meanwhile, TEM was used to observe tight junctions and pyroptosis in the mouse colon. Furthermore, IF, western blotting, RT-qPCR, and ELISA were employed to investigate the mechanism of XXD.

Results: This study indicates that XXD promoted the recovery from dysbiosis and repair of the intestinal barrier. Integrative proteomic and transcriptomic analyses identified the NOD-like receptor signaling pathway as a potential key mechanism, with GBP5 as a possible key protein or gene. In the verification of the prediction results, XXD could significantly inhibit the protein expression of GBP5, NLRP3, ASC, Pro-Caspase1/Cleaved-Caspase1, and N-GSDMD/GSDMD; upregulate the protein levels of ZO-1 and occludin; and reduce the mRNA levels of GBP5 and NLRP3. In addition, it reduced the secretion of IL-1β and IL-18. IF confirmed the co-localization of EpCAM-GSDMD, GBP5-NLRP3, and NLRP3-ASC in colon tissues, whereas TEM suggested that XXD alleviated the ultrastructural damage caused by pyroptosis.

Conclusion: This study demonstrated that XXD can repair the intestinal mucosal barrier and regulate dysbiosis and its associated symptoms. Multiomics approaches have been used to predict the potential mechanisms of XXD. Evidence from experiments indicates that the regulation of dysbiosis by XXD may involve alleviating pyroptosis via inhibition of the GBP5/NLRP3 pathway.