Yuzhi Zhixue Granules regulate the JAK/STAT pathway to affect T-cell differentiation in the colon of ulcerative colitis.

PURPOSE

Yuzhi Zhixue (YZZX) granules, a classic Chinese herbal remedy, have demonstrated efficacy in managing hemorrhagic and inflammatory conditions, and are clinically used to treat uterine bleeding. Building upon their documented clinical applications and phytochemical composition, we hypothesized that YZZX may also possess therapeutic potential against ulcerative colitis (UC). This research aimed to examine the immunomodulatory effects of YZZX granules on Th1/Th17-mediated immune reactions in a murine UC model induced by dextran sulfate sodium (DSS).

METHODS

The major small-molecule constituents and macromolecular nano-components, specifically carbon dots (CDs), in YZZX were identified using UHPLC-QE-MS and other modern analytical techniques. UC was induced in mice via ad libitum access to a 2.5 % DSS solution, with YZZX administered orally. Anti-inflammatory effects were assessed by monitoring disease activity index (DAI) scores, body weight, colon length, ulcer area, and histopathological features. To uncover the mechanistic pathways of YZZX in UC treatment, transcriptomic sequencing, Flow cytometry, ELISA, RT-qPCR and Western blotting were conducted on colon tissue samples.

RESULTS

YZZX significantly mitigated body weight loss, reduced DAI scores, ameliorated histopathological damage, and prevented colon shortening in UC mice. It improved the intestinal barrier function by promoting the upregulation of mucin MUC-2 and tight junction proteins (e.g., claudin, occludin, and ZO-1). Additionally, YZZX restored immune balance by modulating Th17/Treg and Th1/Th2 cell responses through the normalization of cytokine secretion and transcription factor expression. Transcriptomic analysis revealed that YZZX modulates the immune-inflammatory responses in colitis primarily via the JAK/STAT signaling pathway. These findings were further validated by Western blotting and RT-qPCR, which showed that YZZX suppressed the activation of this pathway by downregulating phosphorylated JAK/STAT protein expression.

CONCLUSION

YZZX granules exert immunomodulatory effects primarily by rebalancing Th1/Th2 and Th17/Treg cell populations in the colon via inhibition of the JAK/STAT signaling pathway, thereby alleviating mucosal inflammation. This is the first study to systematically demonstrate the synergistic protective effects of both small-molecule compounds and carbon dot nano-components, derived from a traditional Chinese medicine formula in a DSS-induced murine model of UC.