Modified Pulsatilla decoction alleviates 5-fluorouracil-induced intestinal mucositis by modulating the TLR4/MyD88/NF-κB pathway and gut microbiota

Abstract

BACKGROUND

Modified Pulsatilla decoction (PD), a PD with licorice and ejiao, is a classic Traditional Chinese Medicine formula with significant efficacy in treating intestinal mucositis (IM) induced by tumor therapy. However, its specific molecular and biological mechanisms remain unclear.

AIM

To investigate the therapeutic effect and mechanism of modified PD in IM.

METHODS

This study used an IM mouse model established using 5-fluorouracil injections to investigate the effects of the modified PD (3, 6, and 12 g/kg) in IM. The primary chemical components of the modified PD were identified using liquid chromatography-mass spectrometry. Body weight loss, diarrhea scores, intestinal length, histopathological scores, and inflammatory cytokine levels were measured to evaluate the effects of the modified PD in IM. Effects on the TLR4/MyD88/NF-κB pathway were evaluated using western blot analysis. The intestinal microbiota was characterized using Illumina NovaSeq sequencing.

RESULTS

The results showed that modified PD significantly improved weight loss and diarrhea and shortened the intestines in IM mice. Mechanistically, modified PD suppressed the TLR4/MyD88/NF-κB pathway and downregulated the expression of reactive oxygen species, lipopolysaccharides, and pro-inflammatory cytokines (IL-1β, TNF-α, IFN-γ, IL-6, IL-8, and IL-17), while increasing the expression of the anti-inflammatory cytokine IL-10. Furthermore, modified PD protected the intestinal mucosal barrier by increasing the expression of tight junction proteins (occludin-1, claudin-1, and ZO-1) and mucin-2. Finally, 16S rDNA sequencing revealed that modified PD improved intestinal dysbiosis.

CONCLUSION

Our research offers new insights into the potential mechanism of modified PD in alleviating IM and provides experimental evidence supporting its pharmaceutical application in clinical IM treatment.

Keywords: Modified Pulsatilla decoction; Intestinal mucositis; Oxidative stress; Gut microbiota, TLR4/MyD88/NF-κB pathway; Inflammatory response

Core Tip: Developing safe and effective drugs for the treatment of intestinal mucositis (IM) is of great significance because IM is a common and severe complication of chemotherapy that can lead to a spectrum of gastrointestinal toxicities and even death. In this study, modified Pulsatilla decoction (PD) demonstrated significant efficacy in alleviating 5-fluorouracil-induced IM in mice. The treatment significantly improved key clinical symptoms, including weight loss, diarrhea, and intestinal damage, through modulation of the TLR4/MyD88/NF-κB pathway and restoration of intestinal microbiota balance. These findings provide a novel scientific foundation for the potential clinical application of modified PD in IM management.