Huangqi decoction ameliorated intestinal barrier dysfunction via regulating NF-κB signaling pathway in slow transit...

The development of slow transit constipation (STC) is associated with intestinal barrier damage. Huangqi decoction (HQD) is effective in treating STC, but mechanisms are unclear.

To investigate whether HQD alleviates STC by downregulating the nuclear factor κB (NF-κB) signaling pathway and restoring intestinal barrier function.

KM mice were divided into control, model, and HQD treatment groups. Fresh colonic tissues were collected for single-cell RNA sequencing and spatial transcriptome sequencing. The expressions of claudin-1, mucin 2, and NF-κB P65 proteins were detected by immunohistochemistry. In vitro experiments evaluated the effects of HQD on the LS174T cell line.

HQD improved intestinal motility, restored mucosal epithelium function and morphology. Single-cell RNA sequencing and spatial transcriptome sequencing data showed a reduction in goblet cells, decreased mucin 2 secretion, and activated apoptotic pathways in STC mice. The population of intestinal stem cells was reduced, and proliferation along with Wnt/β-catenin pathways were inhibited. STC also altered the distribution of intestinal cell states, increasing immune-associated Enterocyte_C3. Aberrant NF-κB pathway activation was noted across various cell types. After HQD treatment, NF-κB pathway activity was down-regulated, while cell proliferation pathways were up-regulated, alongside an increase in Enterocyte_C1 related to material transport. Immunocytochemical, Western blot, and immunohistochemistry analyses confirmed NF-κB pathway activation in goblet cells of STC mice, with HQD inhibiting this aberrant activation.

STC involves intestinal mucosal barrier damage. HQD may treat STC by suppressing NF-κB signaling in epithelial cells, restoring intestinal epithelial cell function, and promoting mucosal barrier repair.