Background Quercetin, a flavonoid derived from astragalus and other medicinal plants, exhibits anti-cancer activity; however, its mechanisms in gastric cancer, especially regarding tumor immunity, remain incompletely understood. The aim of this study is to elucidate the mechanism of quercetin against gastric cancer. Methods Quercetin was evaluated in vitro for the effects on proliferation, apoptosis, cell cycle, mitochondrial potential, and morphology in MKN-45, AGS and normal GES-1 cells. Its efficacy and safety were assessed in vivo using a xenograft model. A “gastric cancer cells + T cells” co-culture system was established to examine quercetin's influence on T cell function. Target prediction employed network pharmacology, molecular docking, and molecular dynamics simulations, with validation via PD-L1 knockdown and PI3K overexpression rescue. Results Quercetin significantly inhibited MKN-45 proliferation, induced G0/G1 arrest, and promoted apoptosis, accompanied by mitochondrial damage, loss of membrane potential, downregulation of BCL-2, and upregulation of BAX, CytC, cleaved Caspase-9, and cleaved Caspase-3. In vivo, quercetin reduced tumor growth, decreased Ki-67, increased tumor apoptosis and peripheral CD8+ T cells, with no apparent organ toxicity. In co-culture, quercetin enhanced CD8+ T cell cytotoxicity, increasing granzyme B, perforin, IL-2, and IFN-γ. Mechanistically, quercetin bound PD-L1, reduced its protein expression without affecting mRNA, and suppressed the PI3K-AKT signaling pathway. Conclusion Quercetin exhibited a dual mechanism: it directly suppressed gastric cancer cell proliferation and induced mitochondria-mediated apoptosis, while also enhancing CD8+ T cell cytotoxicity. Its anti-cancer mechanism involved modulation of PD-L1 protein expression and suppression of the PI3K-AKT signaling pathway. These findings supported the feasibility of quercetin as a potential therapeutic candidate for gastric cancer.

Severe acute pancreatitis (SAP) is associated with high morbidity and mortality. Microcirculatory dysfunction is a critical pathological event in this process and a primary contributor to organ failure (OF). Despite the pivotal role of P-selectin in mediating the adhesion of activated platelets and leukocytes to the vascular endothelium, a process central to microcirculatory dysfunction, effective therapeutic interventions for SAP remain limited. Salvia miltiorrhiza , a traditional Chinese medicinal (TCM) herb, possesses well-documented pharmacological properties, including anti-inflammatory, anticoagulant, and microcirculation-improving effects. This review synthesizes recent advances in understanding the bioactive components of Salvia miltiorrhiza , which ameliorate microcirculation by modulating P-selectin expression and activity through mechanisms targeting its transcription, translation, or post-translational activation. Given the current lack of direct evidence in the context of SAP, we synthesized extensive findings from studies on cardiovascular, gastrointestinal, and inflammatory diseases, as well as from relevant acute pancreatitis (AP)/SAP models. These collective data demonstrate that Salvia miltiorrhiza effectively inhibits platelet aggregation, attenuates leukocyte adhesion, mitigates endothelial injury, and improves perfusion. Substantial evidence suggests that the bioactive compounds derived from Salvia miltiorrhiza function as effective agents against microcirculatory dysfunction by targeting P-selectin. Leveraging this well-defined mechanistic pathway and the promising therapeutic efficacy observed in AP/SAP models, targeting P-selectin with Salvia miltiorrhiza ’s bioactive compounds emerges as a compelling novel strategy for SAP-associated microcirculatory dysfunction, laying a groundwork for subsequent validation studies.

Three-Strain Probiotic Combination (Golden Bifid), a probiotic formulation composed of Bifidobacterium longum, Lactobacillus bulgaricus, and Streptococcus thermophilus, is widely used to modulate gut microbiota homeostasis and treat various gastrointestinal disorders. However, the specific molecular mechanisms underlying its therapeutic effects in slow transit constipation (STC) remain incompletely understood. In this study, we demonstrated that Golden Bifid alleviates loperamide-induced constipation by coordinately modulating host transcriptomic profiles, particularly the MAPK and serotonin signaling pathways, and restoring gut microbiota composition and diversity. These multi-omics findings provide novel mechanistic insights into the clinical efficacy of this probiotic combination, which have not been previously elucidated. Using a loperamide (LOP)-induced STC rat model, Golden Bifid was shown significantly increase defecation frequency, fecal water content, and intestinal motility, while improving the pathological damage of colonic tissues. It also elevated the protein expression of c-kit, 5-HT, 5-HT3R, and 5-HT4R in colonic tissue. RNA sequencing identified 1,998 differentially expressed transcripts in Golden Bifid group compared with the LOP group, with 899 upregulated and 1,099 downregulated. These transcripts were enriched in pathways, such as the mitogen-activated protein kinase (MAPK), tumor necrosis factor (TNF) and estrogen signaling pathway. Additionally, 16S rDNA sequencing demonstrated that the Golden Bifid partially restored gut microbiota structure, increased microbial diversity, and reversed the dysbiosis induced by LOP, notably reducing the abundance of Patescibacteria and modulating microbial taxa at both the phylum and genus levels to resemble the gut microbiota composition of the control group. These findings suggest that Golden Bifid alleviate STC by enhancing c-kit and 5-HT signaling, modulating the MAPK signaling pathway and pathway and restoring gut microbiota balance, offering promising therapeutic potential for STC treatment.

Intestinal mucosal barrier injury represents a critical complication of severe acute pancreatitis (SAP) without effective treatment. This study investigated the efficacy, underlying mechanism, and responsible active compounds of the traditional Chinese medicinal prescription Chaihuang Qingyi Huoxue granule (CHQY) in treating SAP-induced intestinal mucosal barrier injury. SAP was established in Sprague-Dawley rats via intra-pancreaticobiliary duct infusion of sodium taurocholate, followed by oral CHQY administration (3.15 g/kg every 6 h for 12 and 24 h). Blood and tissues were harvested to assess the severity of pancreatitis, intestinal mucosal barrier integrity, and extent of inflammatory injury. Intestine-absorbing compounds were identified using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). Our results showed that CHQY treatment effectively mitigated SAP-induced intestinal mucosal injury, as evidenced by improved intestinal epithelial structure, decreased serum levels of intestinal injury markers (d-lactic acid, diamine oxidase, I-FABP, and Zonulin), restored expression of the tight junction protein ZO-1, and reduced serum endotoxin levels. Furthermore, CHQY administration suppressed the expression of proinflammatory mediator HMGB1, its receptor TLR4, and downstream NF-κB signaling in the intestine, leading to downregulated intestinal IL-1β expression and reduced circulating TNF-α and IL-6. UHPLC-HRMS analysis identified 15 intestine-absorbing compounds in CHQY, of which paeoniflorin sulfite and chrysin-7-O-glucuronide independently inhibited TNF-α-induced tight junction loss in IEC-6 cells and mitigated intestinal mucosal barrier injury in SAP rats through suppressing NF-κB signaling. In summary, CHQY ameliorates SAP-induced intestinal mucosal barrier injury by downregulating the proinflammatory HMGB1/TLR4/NF-κB signaling, with efficacy partially attributed to its active compounds paeoniflorin sulfite and chrysin-7-O-glucuronide.

Carbohydrate kinases serve an oncogenic role in several types of cancer; however, the function of FGGY carbohydrate kinase domain containing (FGGY) in colorectal cancer (CRC) remains unknown. The present study investigated the function and possible molecular mechanisms of FGGY in CRC. The results showed that elevated levels of FGGY mRNA and protein were observed in CRC tissues, and a higher expression of FGGY was associated with advanced N stage and reduced overall survival time in patients with CRC. Silencing FGGY inhibited the viability of CRC cells by inducing cell cycle arrest and promoting apoptosis in vitro, thereby attenuating tumor growth in a xenograft mouse model. FGGY knockdown also enriched the senescence‑associated heterochromatin foci (SAHF) pathway and p53 pathway, as further confirmed by enhancing senescence‑associated β‑galactosidase (SA‑β‑gal) activity, with increased levels of SAHF‑associated proteins HP1γ and trimethylation of H3K9 (H3k9me3) in CRC cells, as well as upregulation of p53 and its downstream protein p21. Furthermore, p53 knockout rescued FGGY knockdown‑mediated reductions in cell viability, SA‑β‑gal activity, and the levels of HP1γ and H3k9me3 in CRC cells. These findings indicated that FGGY could act as a newly identified potential oncogene in CRC, partially through regulating the p53/p21 signaling pathway and altering cell senescence.

Background:Raised erosive gastritis (REG) is a chronic gastritis with a high risk of malignant transformation. Current treatments often result in high recurrence rates and complications. Jianpi Qinghua Sanyu Yin (JPQHSYY), a traditional Chinese medicine, shows promise in treating REG. However, the underlying molecular mechanisms remain unclear. This study aimed to investigate the potential mechanism of JPQHSYY's therapeutic effects on REG. Methods:RNA-seq was employed to systematically analyze mRNA, lncRNA, and miRNA profiles in gastric mucosal tissues from REG patients before and after JPQHSYY treatment. The pivotal lncRNA-miRNA and miRNA-mRNA networks were predicted from sequencing data and bioinformatic analysis, and the results were exported using Cytoscape software. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used for functional exploration. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to validate RNA-seq analysis results. CCK8, cell cycle, apoptosis and western blot were performed to detect the effects of miR-122-5p in GES-1 cells in vitro. Results:RNA-seq analysis revealed 576 differentially expressed lncRNAs (269 upregulated, 307 downregulated), 33 differentially expressed miRNAs (13 upregulated, 20 downregulated), and 1717 differentially expressed mRNAs (777 upregulated, 940 downregulated) in JPQHSYY-treated REG patients. GO and KEGG analyses highlighted key pathways, including the PI3K/AKT signaling pathway, involved in cell cycle and apoptosis regulation. The ceRNA network analysis suggested that JPQHSYY impacts the miRNA-lncRNA interactions. Validation experiments confirmed that JPQHSYY inhibits the PI3K/AKT pathway, reducing cell viability, colony formation, and promoting apoptosis in miR-122-5p transfected GES-1 cells. Conclusion:The therapeutic efficacy of JPQHSYY in treating REG might be mediated by the ceRNA-driven PI3K/AKT pathway signaling pathways, which is implicated in the proliferation of gastric mucosal epithelial cells. Furthermore, the investigation of miRNA-lncRNA networks could reveal more information on potential new mechanisms and targets for JPQHSYY in the management of REG.

Stroke is the second leading cause of death worldwide. Organoids, as disease models that closely mimic human physiology and pathology, are highly suitable for investigating the role of neural cells in brain diseases. However, there are few reports on circRNA research based on cerebral organoid stroke models. In this study, we established a human cerebral organoid ischemic stroke model induced by iPSCs and performed transcriptome sequencing. We found that at Days 31 and 52 of organoid development, the microglial marker Iba1, neuronal marker NeuN, and astrocytic marker GFAP exhibited strong fluorescent signals and showed co-localization with the proliferation marker KI67. The organoids showed high levels of the vascular marker CD31 and α-SMA, with co-localization between them. Transcriptome sequencing identified a total of 18 differentially expressed circRNAs (DEcircRNAs) and 2,111 differentially expressed genes (DEGs) in the human cerebral organoid stroke model. KEGG pathway enrichment analysis and WGCNA showed that these DEcircRNAs were involved in the pyroptosis pathway. Furthermore, we identified a pyroptosis-associated circRNA circFGFR2 who was up-regulated in astrocytes and promoted astrocyte pyroptosis. In summary, this study identified a highly expressed pyroptosis-associated circFGFR2 in the human cerebral organoid ischemic stroke model and demonstrated that circFGFR2 promoted astrocyte pyroptosis. This study provides new insights into the pathogenic mechanisms of ischemic stroke.

In this study, loperamide hydrochloride (LOP) was used to establish a rat model of functional constipation (FC) to explore the mechanism of Atractylodes macrocephala insoluble dietary fiber (AMDF) in improving FC. The results showed that AMDF could increase the fecal water content and small intestinal advancement rate of FC rats, raise the levels of gastrin (GAS), vasoactive intestinal peptide (VIP), and substance P (SP) in serum, reduce the level of somatostatin (SS), reduce the levels of IL-6, TNF-α, and NO in serum, and alleviate the pathological state of colon tissue. The 16S rDNA sequencing results revealed that AMDF could enhance the diversity of the intestinal microbiota and restore the ratio of Firmicutes/Bacteroides (F/B). Metabolomics results show that AMDF treatment can regulate bile secretion, primary bile acid biosynthesis, amino acid metabolic signaling pathways, etc. Meanwhile, AMDF could inhibit the activation of the NF-κB signaling pathway in colon tissue and alleviate intestinal inflammation in FC rats. In conclusion, the research results showed that AMDF can improve the intestinal microbiota and metabolism, and inhibit the intestinal inflammatory response, achieving the effect of relieving FC.

Dachengqi decoction (DCQD) is a traditional Chinese medicine formula consisting of three botanical drugs and one mineral drug. It has anti-inflammatory, gastrointestinal motility, and microcirculation improvement effects. A large number of studies have shown that DCQD has a significant impact on the treatment of acute pancreatitis (AP). However, owing to the multi-component and multi-target characteristics of traditional Chinese medicine, it is difficult for modern scientists to understand the role of DCQD in treating AP. Therefore, We sorted out the literature data related to DCQD published in databases such as Web of Science, PubMed, and CNKI, and summarized the main metabolites of DCQD, including anthraquinones, flavonoids, lignans and other metabolites. The pharmacokinetic characteristics of the main metabolites entering the blood circulation are preliminarily summarized. Combining chemical analysis and network analysis, some metabolites in DCQD, such as emodin, rhein, and luteolin, may play important roles in the treatment of AP, and this review also summarizes the efficacy mechanisms of these metabolites. In brief, we have summarized the treatment of AP with DCQD from the aspects of metabolites, preliminary pharmacokinetic analysis, and efficacy mechanisms, to facilitate further understanding of the material basis of DCQD.

Abstract Background Pien Tze Huang (PZH), a traditional Chinese medicine formulation, is recognized for its therapeutic effect on colitis and colorectal cancer. However, its protective role and underlying mechanism in colitis-associated colorectal cancer (CAC) remain to be elucidated. Methods A CAC mouse model was established using AOM/DSS. Twenty mice were randomly divided into four groups (n = 5/group): Control, PZH, AOM/DSS, and AOM/DSS + PZH groups. Mice in the PZH and AOM/DSS + PZH group were orally administered PZH (250 mg/kg/d) from the first day of experiment, while the control and AOM/DSS group received an equivalent volume of distilled water. Parameters such as body weight, disease activity index (DAI), colon weight, colon length, colon histomorphology, intestinal tumor formation, serum concentrations of pro-inflammatory cytokines, proliferation and apoptosis in colon tissue were assessed. RNA sequencing was employed to identify the differentially expressed transcripts (DETs) in colonic tissues and related signaling pathways. Wnt/β-Catenin Pathway-Related genes in colon tissue were detected by QPCR and immunohistochemistry (IHC). Results PZH significantly attenuated AOM/DSS-induced weight loss, DAI elevation, colonic weight gain, colon shortening, histological damage, and intestinal tumor formation in mice. PZH also notably decreased serum concentration of IL-6, IL-1β, and TNF-α. Furthermore, PZH inhibited cell proliferation and promote apoptosis in tumor tissues. RNA-seq and KEGG analysis revealed key pathways influenced by PZH, including Wnt/β-catenin signaling pathway. IHC staining confirmed that PZH suppressed the expression of β-catenin, cyclin D1 and c-Myc in colonic tissues. Conclusions PZH ameliorates AOM/DSS-induced CAC in mice by suppressing the activation of Wnt/β-catenin signaling pathway.

Proteasome inhibition emerges as a promising strategy for cancer prevention. PNO1, pivotal for colorectal cancer (CRC) progression, is involved in proteasome assembly in Saccharomyces cerevisiae. Hence, we aimed to explore the role of PNO1 in proteasome assembly and its up- and down-streams in CRC. Here, we demonstrated that PNO1 knockdown suppressed CRC cells growth, proteasome activities and assembly, as well as CDKN1B/p27Kip1 (p27) degradation. Moreover, p27 knockdown partially attenuated the inhibition of HCT116 cells growth by PNO1 knockdown. The up-stream studies of PNO1 identified miR-326 as a candidate miRNA directly targeting to CDS-region of PNO1 and its overexpression significantly down-regulated PNO1 protein expression, resulting in suppression of cell growth, decrease of proteasome activities and assembly, as well as increasing the stability of p27 in CRC cells. These findings indicated that miR-326 overexpression can suppress CRC cell growth, acting as an endogenous proteasome inhibitor by targeting PNO1.

OBJECTIVE:To explore the regulatory effect of Pien Tze Huang (PZH) on targeting partner of NOB1 (PNO1) and it's down-stream mediators in colorectal cancer (CRC) cells.METHODS:Quantitative polymerase chain reaction was performed to determine mRNA levels of PNO1, TP53, and CDKN1A. Western blotting was performed to determine protein levels of PNO1, p53, and p21. HCT-8 cells were transduced with a lentivirus over-expressing PNO1. Colony formation assay was used to detect cell survival in PNO1 overexpression of HCT-8 cells after PZH treatment. Cell-cycle distribution, cell viability and cell apoptosis were performed to identify the effect of PNO1 overexpression on cell proliferation and apoptosis of HCT-8 cells after PZH treatment. Xenograft BALB/c nude mice bearing HCT116 cells transduced with sh-PNO1 or sh-Ctrl lentivirus were evaluated. Western blot assay was performed to detect PNO1, p53, p21 and PCNA expression in tumor sections. Terminal deoxynucleotidyl transferase dUTP nick end labling (TUNEL) assay was used to determine the apoptotic cells in tissues.RESULTS:PZH treatment decreased cell viability, down-regulated PNO1 expression, and up-regulated p53 and p21 expressions in HCT-8 cells (P<0.05). PNO1 overexpression attenuated the effects of PZH treatment, including the expression of p53 and p21, cell growth, cell viability, cell cycle arrest and cell apoptosis in vitro (P<0.05). PNO1 knockdown eliminated the effects of PZH treatment on tumor growth, inhibiting cell proliferation inhibition and apoptosis induction in vivo (P<0.05). Similarly, PNO1 knockdown attenuated the effects of PZH treatment on the down-regulation of PNO1 and up-regulation of p53 and p21 in vivo (P<0.05).CONCLUSION:The mechanism by which PZH induces its CRC anti-proliferative effect is at least in part by regulating the expression of PNO1 and its downstream targets p53 and p21.

Slow transit constipation (STC) is a common and debilitating condition characterized by delayed colonic transit and difficulty in fecal expulsion, significantly impacting patients’ physical and mental wellbeing as well as their overall quality of life. This study investigates the therapeutic potential of Liqi Tongbian Decoction (LTD) in the treatment of STC, especially in cases involving the context of Qi stagnation, through a multifaceted approach involving the modulation of intestinal flora and short-chain fatty acids (SCFAs). We employed a rat model of STC with Qi Stagnation Pattern, established using the “loperamide + tail-clamping provocation method,” to explore the effects of LTD on fecal characteristics, intestinal motility, and colonic pathology. Importantly, LTD exhibited the ability to increase the richness, diversity, and homogeneity of intestinal flora while also modulating the composition of microorganisms. It significantly increased the production of SCFAs, especially butyric acid. Moreover, LTD exerted a substantial influence on the synthesis of serotonin (5-HT) by modulating the expression of tryptophan hydroxylase (TPH) and interacting with the 5-HT4 receptor (5-HT4R), resulting in enhanced colonic motility. Correlation analyses revealed a positive correlation between certain bacterial genera, such as Lachnospiraceae_NK4A136 spp. and Clostridiales spp. and the concentrations of butyric acid and 5-HT. These results suggest a mechanistic link between microbiome composition, SCFAs production, and 5-HT synthesis. These findings highlight the potential of LTD to alleviate STC by facilitating a beneficial interplay among intestinal flora, SCFAs production, and 5-HT-mediated colonic motility, providing novel insights into the management of STC with Qi Stagnation Pattern.

ContextUlcerative colitis has been clinically treated with Qing Hua Chang Yin (QHCY), a traditional Chinese medicine formula. However, its precise mechanisms in mitigating chronic colitis are largely uncharted.ObjectiveTo elucidate the therapeutic efficiency of QHCY on chronic colitis and explore its underlying molecular mechanisms.Materials and methodsA total ion chromatogram fingerprint of QHCY was analysed. Chronic colitis was induced in male C57BL/6 mice using 2% dextran sodium sulphate (DSS) over 49 days. Mice were divided into control, DSS, DSS + QHCY (0.8, 1.6 and 3.2 g/kg/d dose, respectively) and DSS + mesalazine (0.2 g/kg/d) groups (n = 6). Mice were intragastrically administered QHCY or mesalazine for 49 days. The changes of disease activity index (DAI), colon length, colon histomorphology and serum pro-inflammatory factors in mice were observed. RNA sequencing was utilized to identify the differentially expressed transcripts (DETs) in colonic tissues and the associated signalling pathways. The expression of endoplasmic reticulum (ER) stress-related protein and NF-kappa B signalling pathway-related proteins in colonic tissues was detected by immunohistochemistry staining.ResultsForty-seven compounds were identified in QHCY. Compared with the DSS group, QHCY significantly improved symptoms of chronic colitis like DAI increase, weight loss, colon shortening and histological damage. It notably reduced serum levels of IL-6, IL-1 beta and TNF-alpha. QHCY suppressed the activation of PERK-ATF4-CHOP pathway of ER stress and NF-kappa B signalling pathways in colonic tissues.Discussion and conclusionsThe findings in this study provide novel insights into the potential of QHCY in treating chronic colitis patients.

Table S6 indicates that PNO1 mRNA expression is significantly up-regulated in various types of malignancies, including CRC, lung cancer, and breast cancer.

5-Fluorouracil (5-FU) is one of the most commonly prescribed anticancer agents. However, intestinal mucositis is a well-known adverse event, which limits its therapeutic use. Refined Baohe formula (RBF) is derived from Baohe Pills with a potent gastrointestinal protective effect. However, the effects of RBF on 5-FU-induced intestinal mucositis remain unknown. Thus, this study investigated the effects and mechanism of RBF on 5-FU-induced intestinal mucositis in a CT-26 xenograft mice colorectal cancer (CRC) model. The composition of the RBF preparation was analyzed by high-pressure liquid chromatography. CT-26 cells bearing mice were intraperitoneally administered 150 mg/kg of 5-FU on day 1 to construct a 5-FU-induced intestinal injury model. Mice in the 5-FU + RBF group were intragastrically administrated with RBF (12.4 g/kg) for 4 days. Tumor volume and weight were determined, body weight and diarrhea scores were monitored, leukocytes were calculated, and hematoxylin-eosin staining, immunohistochemistry, and terminal deoxynucleotidyl transferase dUTP nick end labeling were performed to investigate the therapeutic efficiency and underlying mechanisms of RBF on 5-FU-induced intestinal mucositis mice. RBF didn`t affect the effect of tumor suppression, while attenuated diarrhea associated with 5-FU-induced intestinal mucositis and significantly reduced leucocyte toxicity. Additionally, RBF significantly promoted cell proliferation and inhibited cell apoptosis, and up-regulated CDK4, c-Myc, and B-cell lymphoma-2 expression, while down-regulated Bax expression, as well as enhanced both p-AKT and AKT expression in intestinal crypts of 5-FU-treated mice. RBF attenuated 5-FU-induced intestinal mucositis by inhibiting apoptosis and promoting cell proliferation by modulating the AKT pathway and its downstream effectors in a CT-26 xenograft mouse CRC model.

Table S2 shows the double-stranded shRNAs targeting selected up-regulated genes and siRNA for PNO1.

Background: Qing Hua Chang Yin (QHCY) is a famous formula of traditional Chinese medicine (TCM) and has been proven to have protective effect on ulcerative colitis. However, its protective effect and potential therapeutic mechanisms in chronic colitis remain unclear. The purpose of this study is to explore the effects and underlying mechanisms of QHCY on dextran sulfate sodium (DSS)-induced chronic colitis mice model. Methods: The chronic colitis model was established by administration of 2% DSS for three consecutive cycles of 7 days with two intervals of 14 days for recovery by drinking water. The experiment lasted 49 days. The DSS + QHCY group received QHCY administration by oral gavage at doses of 1.6 g/kg/d, DSS + Mesalazine group was administrated Mesalazine by oral gavage at doses of 0.2 g/kg/d. The control and DSS group were given equal volume of distilled water. The body weight, stool consistency and blood in stool were monitored every 2 days. The disease activity index (DAI) was calculated. The colon length was measured after the mice were sacrificed. The histomorphology of colonic tissues was checked by the HE and PAS staining. Immunohistochemistry was performed to detect the expressions of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6), tight junction proteins (ZO-1, occludin) and Mucin2 (MUC2). 16S rRNA sequencing analysis was conducted to study the diversity and abundance of gut microbiota changes. Results: QHCY treatment not only significantly attenuated DSS-induced the weight loss, DAI score increase, colon shortening and histological damage in mice, but also decreased the expression of pro-inflammatory cytokines in colonic tissues and increased the expression of ZO-1, occludin, and MUC2. Furthermore, QHCY enhanced the diversity of gut microbes and regulated the structure and composition of intestinal microflora in mice with chronic colitis. Conclusion: QHCY has a therapeutic effect on a murine model of chronic colitis. It can effectively reduce the clinical and pathological manifestations of colitis and prevent alterations in the gut microbiota.

Table S3 summaries primer sequences and probe for quantitative PCR or Northern blotting