Therapeutic potential of Qihuang Biwen Formula and its bioactive compounds for the treatment of ulcerative colitis in Drosophila and mice.

Abstract

BACKGROUND: Qihuang Biwen Formula (QHBWF), derived from the classic Yupingfeng Powder, is traditionally used for immune regulation. While clinically applied, its mechanism of action in ulcerative colitis (UC) remains unclear. PURPOSE: The effectiveness and underlying principles of QHBWF for UC treatment were investigated using both Drosophila and mice models, and its bioactive components were identified. METHODS: In the dextran sulfate sodium (DSS)-induced adult fly, the survival rate, locomotion, excretion, smurf, digestive capacity, intestinal length, death of intestinal epithelial cells (IECs), and proliferation of intestinal stem cells (ISCs) were measured. Meanwhile, the body weight, intestinal length, disease activity index, histopathology, serum cytokine levels, and ROS levels were detected in the UC mouse. Additionally, the mechanism of QHBWF was investigated by bioinformatics analysis, transcriptomic analysis, qRT-PCR, immunohistochemistry and western blotting (WB). Finally, UCPL-MS/MS technology and phenotype assays in UC flies were used to screen the key bioactive components of QHBWF. RESULTS: QHBWF significantly improved survival rate, locomotor activity, and intestinal function. It restored intestinal morphology, suppressed aberrant intestinal stem cell (ISC) proliferation, and reduced intestinal epithelial cell (IEC) apoptosis in DSS treated flies. In mice, QHBWF alleviated weight loss, colon shortening, and DAI scores, improved histopathology, and reduced IL-6, IL-1β, and intestinal ROS. These results indicated that QHBWF could alleviate the intestinal injury in both UC models. Mechanistically, QHBWF down-regulated mRNA levels of Toll signaling and the Imd signaling, and up-regulated the gene expression of mitochondrial oxidative phosphorylation (OXPHOS) complexes in the intestines of UC flies. Consistently, QHBWF decreased the protein levels of TLR4, MyD88 and NF-κB p65, and promoted the protein expresses of NDUFB8, SDHB and ATP5A in the guts of UC mice. These indicated that QHBWF inhibited the TLR4/NF-κB signaling pathway and activated the OXPHOS function. Furthermore, kaempferol, emodin, quercetin, isochlorogenic acid, and chlorogenic acid could extend the survival rate, increased the intestinal length, and facilitated intestinal barrier repair in UC flies. CONCLUSION: QHBWF exerted anti-colitis effects by regulating immunity and metabolism, targeting the TLR4/NF-κB signaling pathway and mitochondrial oxidative phosphorylation. Kaempferol, emodin, quercetin, isochlorogenic acid, and chlorogenic acid were the bioactive compounds of QHBWF against UC. This study provides the potential of QHBWF as a multi-target treatment for UC.