Enhanced therapeutic effect on colitis with powder formulations of Painong San associated with the promotion of intestinal adhesion and absorption.

ETHNOPHARMACOLOGICAL RELEVANCE: Painong San is a prescription composed of traditional Chinese medicine, which has been used to treat colitis. The Painong San's usage recorded in "Jingui Yaolve" by Zhongjing Zhang in the Later Han Dynasty is powder. However, the decoction is often used in reality. It's unclear which dosage form of Painong San is more suitable for colitis treatment and why? AIM OF THE STUDY: This study aims to evaluate the different therapeutic effects of Painong San (a powder of Painong San) and Painong Decoction (a decoction of Painong San) on a dextran sulfate sodium salt-induced colitis model and the possible reasons of these different effects. MATERIALS AND METHODS: The contents of paeoniflorin, naringin, heperidin and neohesperidin in Painong San and Painong decoction were determined by an ultra-performance liquid chromatography system. The therapeutic effect on colitis was evaluated by intragastric administration of Painong San or Painong Decoction in dextran sulfate sodium salt-induced mouse model. The accumulated release rate of Painong San in vitro was analyzed with artificial gastric juice, artificial intestinal juice, and artificial colon juice. The concentrations of four compounds in the blood, intestinal contents, and intestinal fluids were detected by an-ultra performance liquid chromatography-tandem quadrupole mass spectrometry system. In situ intestinal perfusion experiments were used to observe the adhesion of Painong San to the intestine's surface. The expression of Mucin-2 and Trefoil Factor 3 in the colon was studied by immunohistochemistry. RESULTS: There was no significant difference in the contents of the four compounds in Painong San and Painong Decoction. In vivo, Painong San has a better therapeutic effect than Painong Decoction in the treatment of colitis. Painong San could be released slowly in the simulated human digestion environment in vitro, and more Painong San particles were released on the intestinal surface in the colitis state in the healthy state. Painong San could increase the bioavailability of hesperidin and neohesperidin, and their concentrations in local intestinal tissue, intestinal fluid and intestinal contents. The expression of trefoil Factor 3 protein on the surface of the colon tended to be in the intestinal cavity and overlapped with the expression of mucin-2. Trefoil Factor 3 accumulates in the intestinal cavity of the colon in the state of colitis, which may increase the adhesion of Painong San particles. CONCLUSION: In conclusion, this experiment proved that Painong San is a more suitable dosage form for the treatment of dextran sulfate sodium salt-induced colitis than Painong Decoction, which may be related to the enhancement of Painong San particle adhesion to the intestine in colitis. This study provides a reference for the selection of clinical dosage forms for traditional Chinese medicine.

Painong San, a Traditional Chinese Compound Herbal Medicine, Restores Colon Barrier Function on DSS-Induced Colitis in Mice.

OBJECTIVE: The intestinal barrier decreases in colitis and restores the integrity of the mucosal barriers that could be used for the treatment of colitis. Painong San (PNS), a traditional Chinese compound herbal medicine originally recorded in "Jingui Yaolve" by Zhongjing Zhang in the Later Han Dynasty, is often used in China and Japan to treat various purulent diseases including intestinal carbuncle. This study was to investigate the effect of PNS on mucosal barrier function in mice with DSS-induced colitis and its related mechanisms. METHODS: BALB/C mice were given 3% DSS to induce colitis. The body weight and stool status of the mice were recorded daily, and the histopathological changes of the colon were observed after execution. The permeability of the intestinal mucosa was measured by fluorescein isothiocyanate-dextran 4000, the change of intestinal microbiota was measured by 16S rDNA, and the tight junction-related proteins and Muc-2 were investigated by immunohistochemical or immunofluorescence. The possible signaling pathways were detected by western blot. RESULTS: Compared with the control group, the composition of the microbiota in the PNS group was close to that of the normal group, the number of goblet cells was improved, and the mucosal permeability was significantly reduced. PNS could upregulate the expression of tight junction-related proteins (ZO-1, claudin-1, and occludin) and Muc-2, and at the same time, regulate the Notch pathway. CONCLUSION: PNS could effectively improve the mucosal barrier function through multiple ways, including restoring the balance of intestine flora, enhancement of the mucous layer barrier, and mechanical barrier function. These protective effects may relate to inhibiting the Notch signaling pathway activated by DSS.

Protective effect of Pai-Nong-San against AOM/DSS-induced CAC in mice through inhibiting the Wnt signaling pathway.

Pai-Nong-San (PNS), a prescription of traditional Chinese medicine, has been used for years to treat abscessation-induced diseases including colitis and colorectal cancer. This study was aimed to investigate the preventive effects and possible protective mechanism of PNS on a colitis-associated colorectal cancer (CAC) mouse model induced by azoxymethane (AOM)/dextran sodium sulfate (DSS). The macroscopic and histopathologic examinations of colon injury and DAI score were observed. The inflammatory indicators of intestinal immunity were determined by immunohistochemistry and immunofluorescence. The high throughput 16S rRNA sequence of gut microbiota in the feces of mice was performed. Western blot was used to investigate the protein expression of the Wnt signaling pathway in colon tissues. PNS improved colon injury, as manifested by the alleviation of hematochezia, decreased DAI score, increased colon length, and reversal of pathological changes. PNS treatment protected against AOM/DSS-induced colon inflammation by regulating the expression of CD4+ and CD8+ T cells, inhibiting the production of HIF-α, IL-6, and TNF-α, and promoting the expression of IL-4 and IFN-γ in colon tissues. Meanwhile, PNS improved the components of gut microbiota, as measured by the adjusted levels of Firmicutes, Bacteroidetes, Proteobacteria, and Lactobacillus. PNS down-regulated the protein expression of p-GSK-3ß, ß-catenin, and c-Myc, while up-regulating the GSK-3ß and p-ß-catenin in colon tissues of CAC mice. In conclusion, our results suggested that PNS exhibits protective effect on AOM/DSS-induced colon injury and alleviates the development of CAC through suppressing inflammation, improving gut microbiota, and inhibiting the Wnt signaling pathway.