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. 2026 Apr 13;17:1767221. doi: 10.3389/fmicb.2026.1767221

Preventive effect of ferulic acid on dextran sulfate sodium-induced ulcerative colitis in mice

Zheng Zhong 1, Zhihan Dong 2, Hongya Zheng 1, Yanzhu Zhu 2,3, Caoxing Huang 4, Wei Lian 5, Baishuang Yin 1,2,*
PMCID: PMC13111449  PMID: 42052396

1. Introduction

Ulcerative colitis (UC) is a chronic, lifelong, non-specific intestinal inflammatory disease, which is charactered with abdominal pain, chronic diarrhea, and rectal bleeding. The incidence of UC in emerging industrialized countries and regions such as Asia, South America and Eastern Europe has shown a rapid growth trend (Ashique et al., 2023; Yamazaki et al., 2023). While the precise etiology of UC remains elusive, the most accepted pathogenesis involves chronic intestinal inflammation. Intestinal microbiota played an important role in the pathogenesis and progression of UC, including decreased microbial diversity, reduced beneficial bacteria and increased harmful bacteria (Wu et al., 2023). While current anti-inflammatory therapies aim to control symptoms, many patients experience drug resistance and low responsiveness. Prednisone (Pdn) is a synthetic glucocorticoid and one of the key medications for treating UC. Although inflammation symptoms could be alleviated by glucocorticoids, metabolic disorders, endocrine abnormalities, hepatic injury, and severe pancreatitis were often observed (Di Rienzo et al., 2024). It is necessary to explore safe and effective natural active ingredients for UC as potential intervention strategies.

Ferula is a resin derived from the plants Ferula Xinjiang or Ferula fukang of the Apiaceae family (Hu et al., 2024). Historically, Ferulic acid (FA) was first isolated and identified from the components of such plants, and thus it got its name (Martínková et al., 2023). It exhibited anti-inflammatory, antioxidant, anticancer, and antibacterial effects (Zhang et al., 2022; Zhang et al., 2023). FA enhanced autophagy, inhibited the activation of NLRP3 inflammasome and reduced the expression and release of inflammatory factors (Liu et al., 2022). FA had a gastroprotective effect against indomethacin-induced gastric ulcers in rats due to its antioxidant and anti-inflammatory properties (Ermis et al., 2023). FA inhibits the inflammatory injury of endothelial cells in 2,4,6-triabrobenzene sulfonic acid-induced UC (Yu S. et al., 2023). However, its role in the other UC model is still unclear.

In this study, the non-infectious UC in mice was established by dextran sulfate sodium (DSS). The protective effect of FA on DSS-induced UC was explored through colonic morphology, inflammatory mediators, antioxidant capacity, and gut microbiota compared with the Pdn. It will provide a theoretical basis for the expanded application of FA in UC prevention.

2. Materials and methods

2.1. Reagents

The molecular weight of DSS is 36–50 kda and it was purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. Item No. 60316ES76. Pdn was purchased from Shanghai Maokang Biotechnology Co., Ltd. Item number: MZ10301-25G. Qinglong PI is extracted from the green husks of walnuts in Changbai Mountain (jilin) by Nanjing Forestry University.

2.2. Experimental animal

Sixty male Kunming mice (6–8 weeks old and weighing 20 ± 2 g) were housed in ventilated cages under controlled conditions (23 ± 2 °C; 50–60% humidity; 12-h light/dark cycle). The standard food and water were available ad libitum, and bedding were replaced every other day. After a 7-day acclimatization, the mice were randomly allocated to six groups (n = 10, oral gavage): Control group (CON), Dextran sulfate sodium group (DSS, 3%), FA group (80 mg/kg), FA+DSS group, Pdn (4 mg/kg), Pdn+DSS group (Figure 1A). The diagram illustrated the experimental flowchart of this study (Figure 1A). DSS (3%) gavage was freshly prepared daily to the mice in the DSS, FA+DSS, and Pdn+DSS groups for 7 days. On the third day before DSS exposure, FA (extracted from Changbai Mountain Qinglongyi) was administered by gavage once daily for 10 days. On the fourth day before DSS exposure, Pdn was given by gavage once daily for 3 days. Body weight was recorded daily during experiment. At the end of the experiment, mice were euthanized under isoflurane anesthesia. Whole blood was collected from the retro-orbital plexus and centrifuged at a speed of 1, 400 × g for 10 min to obtain serum. The entire colon was excised, and gently rinsed with ice-cold normal saline to remove luminal contents without damaging the mucosa. Colon segments (2 cm) were immobilized in a 4% formaldehyde solution. Colon samples (1 mm3) were immediately immersed in ice-cold 2.5% glutaraldehyd. The dose of DSS was chosen according to Lu’s experiment (Lu et al., 2022). The doses of FA and Pdn was chosen according to Huang and Mencarelli’s experiment, respectively (Huang et al., 2025; Mencarelli et al., 2016).

Figure 1.

Panel A shows a timeline diagram illustrating experimental groups of mice given different oral treatments (physiological saline, ferulic acid, prednisone) with or without dextran sulfate sodium over seventeen days, ending in sacrifice. Panel B displays a line graph comparing weight changes across experimental groups over seven days. Panel C presents a bar graph of disease activity index among groups, with significance indicated by asterisks. Panel D contains four photographs of mice representing each group (CON, DSS, FA+DSS, Pdn+DSS), highlighting their physical condition and fecal samples.

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The clinical symptom evaluation (n = 10). (A) Experimental flowchart, (B) body weight of mice in each group was monitored daily for 7 days, (C) DAI disease score was calculated, (D) the main symptoms of UC were captured.

2.3. Detection of disease activity index (DAI)

The Disease Activity Index (DAI) scores in the mice were calculated according to DAI method (Zhang et al., 2024). The Disease Activity Index (DAI) scores were observed as follows: (a) weight loss: 0 points = not available, 1 point = 1–5%, 2 points = 5–10%, 3 points = 10–15%, and 4 points = more than 15%; (b) fecal consistency: 0 point = normal, 1 point = not rigorous, 2 points = soft stool, 3 points = meager stool, and 4 points = constipation; (c) hemorrhage: 0 points = normal, 1 point = negative fecal occult blood count, 2 points = positive for occult blood, 3 points = obvious bloody stool, and 4 points = profuse bleeding.

2.4. Histopathological examination of mouse colon

Colon segments (2 cm) were immobilized in a 4% formaldehyde solution. A standard ethanol–xylene gradient was used for dehydration and clearing before paraffin infiltration. Fixed colon tissue was embedded in Paraffin. Serial 5 μm sections were cut on a rotary microtome, mounted on glass slides, and stained with hematoxylin and eosin method. Slides were covers with mounting medium and examined under a light microscope (Zhang et al., 2021). The histological scoring was evaluated by two independent blinded investigators.

2.5. The ultrastructural changes of colon tissue

Colon samples (1 mm3) were immediately immersed in ice-cold 2.5% glutaraldehyde for 2 h. The tissue was post-fixed with 1% osmium tetroxide for 2 h. Specimens were dehydrated through graded ethanol solutions, infiltrated with epoxy resin (Epon 812), and polymerized at 60 °C for 48 h. Ultrathin sections (70 nm) were cut on an ultramicrotome, mounted on copper grids, and contrast-stained with uranyl acetate followed by lead citrate. Ultrastructural changes in colon were examined with a transmission electron microscope (Zhu et al., 2016). And the mitochondria histological scoring was evaluated by two independent blinded investigators.

2.6. Detection of colitis factors in mice

Serum levels of IL-1β (YJ301814), IL-6 (YJ098430), IL-2 (YJ35348), and IL-10 (YJ34730) were quantified using ELISA kits (Shanghai Yuanju Biotechnology Center, Shanghai, China) according to the manufacturer’s instructions. Additionally, the levels of lactoferrin (FL) (ml821520) and fecal calprotectin (FC) (ml106769L) in mouse feces were measured using ELISA kits (Shanghai Enzyme-linked Biotechnology Co., Ltd., Shanghai, China).

2.7. Detection of antioxidant factors in mice

To evaluate the oxidative stress and antioxidant defense status of the experimental subjects, the enzymatic activities of superoxide dismutase (SOD, kit catalog no. A001-3-2), glutathione peroxidase (GSH-Px, kit catalog no. A005-1-2), catalase (CAT, kit catalog no. A007-1-1), and total antioxidant capacity (T-AOC, kit catalog no. A015-2-1), as well as the content of malondialdehyde (MDA, kit catalog no. A003-2-2) in the collected serum samples, were quantitatively determined using commercially available assay kits. All these kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu Province, China), and the entire detection process was strictly performed in accordance with the detailed standard operating protocol.

2.8. Western blot analysis

The protein expression of NLRP3 was examined using western blot (Zhu et al., 2020). The supernatants of the colon were collected and determined using a BCA Protein Assay Kit (Beyotime Bio, Beijing, China). 40 μg of protein was separated on 10% sodium dodecyl sulfate–polyacrylamide gels, subjected to electrophoresis and transferred to PVDF membranes (polyvinylidene difluoride). The PVDF membranes were blocked with 5% non-fat milk for 2 h at room temperature, followed by incubation overnight with primary antibodies of NLRP3 (1:2000, 68,102-1-Ig, Proteintech Group, Inc., Wuhan, China) and β-actin (1:5000, 20,536-1-AP, Proteintech Group, Inc., Wuhan, China), on a shaker at 4 °C. Then, the membranes were washed three times with TBS-T and incubated with anti-rabbit horseradish peroxidase-conjugated IgG (1:5000, R00001, Nature Biosciences, Hangzhou, China) for 2 h at room temperature. Subsequently, PVDF were washed three times with TBS-T. The immunoreactivity was detected using an enhanced chemiluminescence reaction. The density of the bands was quantified by Image J version 2.0 (USA). β-actin was used as an internal control to normalize protein loading, and the relative expression of NLRP3 was calculated as the ratio of NLRP3 band density to β-actin band density.

2.9. Gut microbiota sequencing

Fresh luminal contents were aseptically collected from the rectum of each group immediately after euthanasia. Microbial genomic DNA was isolated using the QIAamp Fast DNA Stool Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocol. Gut microbiota composixxtion was analyzed by sequencing of the 16S rRNA V4 gene region. The primers used were 515F (GTGCCAGCMGCCGCGGTAA) and 806R (GGACTACHVGGGTWTCTAAT). Sequencing was conducted on the NovaSeq 6,000 platform (Illumina) using 2 × 250-bp paired-end sequencing with the NovaSeq 6,000 SP Reagent Kit (Illumina) (500 cycles).

2.10. Statistical analysis

The results were analyzed by one-way analysis of variance followed by Tukey’s HSD (SPSS 20.0 software; SPSS Inc., Chicago, IL, USA). The histograms were drawn by the GraphPad Prism (version 8.0, GraphPad Software Inc., San Diego, CA, USA). The results were expressed as mean ± standard deviation (SD). SD was shown on the top of each column. ***p < 0.001 indicated extremely significant difference, **p < 0.01 indicated markedly significant difference, *p < 0.05 indicated significant difference, and p > 0.05 was considered not statistically significant.

3. Experimental results

3.1. Animal model analysis

The average body weight increased in the FA+DSS group (Figure 1B). Seven days after DSS induced exposure, diminished food and water intake, increased irritability, weight loss, and mucoid or bloody diarrhea were documented. The Disease Activity Index (DAI) is a standardized quantitative scoring system used to assess the severity of UC. Based on these symptoms, the DAI disease scores in the DSS modeling exhibited increased trend, while a reduction in DAI disease scores was observed in the FA and Pdn group (Figure 1C). Compared with the CON, the mice in the DSS group had obvious hematochezia conditions, while the hematochezia conditions of the mice in the FA and the Pdn prevention group were significantly alleviated (Figure 1D). As indicated by the above results, the DSS modeling was successful in this experiment.

3.2. Inflammatory cytokines in mice

FC and FL are standard non-invasive biomarkers for the clinical diagnosis of UC. IL-1β, IL-2, IL-6, and IL-10 plays the core roles of initiating inflammation balance. Compared with the DSS group, such increases of FC and FL were significantly suppressed by preventive supplementation with FA and Pdn (p < 0.01, Figures 2A,B). This increase of IL-6 and IL-1β was markedly reduced when FA and Pdn were preventively supplemented (p < 0.01, Figures 2C,D). The level of IL-2 and IL-10 was increased by preventive supplementation with both FA and Pdn (p < 0.05, Figures 2E,F). These results indicate that preventive supplementation of FA and Pdn can effectively alleviate inflammatory responses, effectively preventing UC.

Figure 2.

Eight-panel scientific figure presents six bar graphs (A-F, H) and one western blot image (G). Bar graphs show statistical comparisons of calprotectin, lactoferrin, IL-1β, IL-6, IL-2, IL-10, and NLRP3 levels across control and experimental groups, indicating significance with asterisks and 'ns' for no significance. Western blot in panel G displays NLRP3 and β-actin protein bands for each group, with molecular weights labeled.

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Preventive FA supplementation on the inflammation cytokine and anti- inflammation cytokine (n = 10 per group). FC, FL, IL-1β, IL-6, IL-2, and IL-10 were quantified using ELISA kits. (A) FC, (B) FL, (C) the IL-1β level, (D) the IL-6 level, (E) the IL-2 level, (F) the IL-10 level, (G) NLRP3 protein expression, and (H) mean gray value analysis of NLRP3 protein expression. All data were expressed as the mean ± SD. **p < 0.01, *p < 0.05, and ns p > 0.05. SD was shown on the top of each column.

3.3. NLRP3 protein expression in colon of mice

In UC, the NLRP3 inflammasome is a core inflammatory sensor and amplifier, and its excessive activation is a key link driving intestinal immune imbalance and tissue damage. As presented in Figures 2G,H, increase trend of the NLRP3 protein expression were reduced in the FA+DSS group and the Pdn+DSS group (p < 0.05). This suggests that preventive FA and supplementation can inhibit NLRP3 protein expression in the DSS group.

3.4. Antioxidant function in serum of mice

SOD, GSH-Px, CAT, T-AOC and MDA are precisely the key indicators for evaluating the antioxidant defense capacity and the degree of oxidative damage. Compared with the DSS group, preventive FA and Pdn supplementation significantly increased the activities of SOD, GSH-Px, CAT, T-AOC, and reduced the MDA content (p < 0.01, p < 0.05, Figures 3A–E). From the above results, preventive administration of FA and Pdn can alleviate the oxidative damage induced by DSS.

Figure 3.

Bar graph with five panels labeled A to E, each comparing treatment groups for antioxidant or oxidative stress markers: CAT, GSH-PX, SOD, T-AOC, and MDA. Axes display marker concentration; statistical significance is indicated by asterisks and “ns” marks above bars, comparing CON, DSS, FA+DSS, FA, Pan, and Pan+DSS groups. Error bars show standard deviation.

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Preventive FA supplementation on the anti-oxidative function (n = 10 per group). The activity of SOD, GSH-Px, CAT, T-AOC activity, and MDA content were measured using Commercial kits. (A) The CAT activity, (B) the GSH-Px activity, (C) the SOD activity, (D) the T-AOC activity, (E) the MDA content. All data were expressed as the mean ± SD. **p < 0.01, *p < 0.05, and ns p > 0.05. SD was shown on the top of each column.

3.5. Colon histopathology in mice

Colon histopathology with HE staining serves as the standard and ultimate evidence for evaluating the success of UC models. The extensive inflammation infiltration and crypt destruction were observed in the DSS group (Figures 4A,B). The colon injury was substantially attenuated in the FA+DSS group and Pdn+DSS groups (Figures 4C,D). The Colonic histological injury score was increased in DSS group, and it was lower in the FA+DSS group and Pdn+DSS group (Figure 4E) (p < 0.01) This study demonstrates that prophylactic administration of FA and Pdn can effectively alleviate the Colon histopathological damage in mice with DSS-induced colitis.

Figure 4.

Panel A to D shows microscopic images of colon tissue with varying degrees of cellular damage. Panel E is a bar graph comparing colonic histological injury scores among four groups, showing significantly higher injury in the DSS group compared to FA+DSS and Pun+DSS groups, with statistical significance indicated by double asterisks.

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Histopathology of mice colon (n = 10 per group, 50 μm). The histopathology of mice colon was determined by HE staining method. (A) CON, (B) DSS group, (C) FA+DSS group, (D) Pdn+DSS group, (E) Colonic histological injury score. All data are expressed as the mean ± SD. **p < 0.01. SD was shown on the top of each column.

3.6. Gut microbiota diversity in mice

The diversity of the intestinal microbiota using 16S rRNA sequencing method is of vital importance in UC. This disorder can directly trigger or exacerbate UC. Muribaculaceae and Clostridia was at a much higher level in the DSS group than that in the CON. A notable decrease was observed in the relative abundance of beneficial genera such as Lactobacilli, Bifidobacteria, Bacteroides in the DSS group. Following preventive administration of FA and Pdn, a restoration of the intestinal microbial community was observed, as evidenced by a significant increase of beneficial bacteria. Muribaculaceae and Clostridia were reduced, Bacteroides was increased in the FA+DSS and Pdn+DSS groups, indicating that FA and Pdn exerted a regulatory effect on gut microbiota composition. Based on these findings, it can be concluded that preventive administration of FA and Pdn improves the gut microbiota in mice (Figure 5).

Figure 5.

Panel A shows a stacked bar chart of relative genus abundance across samples grouped by six treatment conditions, with a color-coded genus legend. Panel B contains a PCA plot with colored clusters for each group, showing separation of microbial communities, and side boxplots. Panel C presents a heatmap representing enzyme abundance patterns across the same sample groups. Panel D displays a petal diagram indicating shared and unique operational taxonomic units among the six groups, with a core of two hundred fifty shared units and petal-wise unique counts.

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Gut microbiota diversity in mice (n = 3 per group). Gut microbiota composition was analyzed by sequencing of the 16S rRNA V4 gene region. C, CON, D, DSS group, FD, FA+DSS group, F, FA group, P, Pdn group, FU, Pdn+DSS group. (A) Community bar chart, (B) PCA graph, (C) Heatmap of Enzyme, (D) Venn diagram. C, CON, D, DSS group, FD, FA + DSS group, F, FA group, P, Pdn group, FU, Pdn+ DSS group.

3.7. The ultrastructural colon in mice

The ultrastructural colon is key supplementary tools for revealing the damage of UC. As shown in Figure 6, In the DSS group, mitochondria were partially swollen with extravasated contents, and the marginalization of chromatin and endoplasmic reticulum dilation were observed. In the FA+DSS group and Pdn+DSS group, the lesion was alleviated. The mitochondria damage score was lower in the FA+DSS group and Pdn+DSS group compared the CON (p < 0.01). The Mitochondrial score was increased in DSS group, and mitochondria damage score was lower in the FA+DSS group and Pdn+DSS grou (p < 0.01) These findings indicate that FA and Pdn attenuate DSS-induced mitochondrial damage.

Figure 6.

Panel A to D contain grayscale transmission electron micrographs of cellular structures labeled with red and blue arrows, highlighting mitochondrial and nuclear features. Enlarged sections illustrate mitochondrial morphology. Scale bars indicate magnification levels. Panel E presents a vertical bar chart labeled "Mitochondrial damage score" on the y-axis and groups "CON," "DSS," "FA+DSS," and "Pdrn+DSS" on the x-axis, showing significant differences marked with double asterisks between groups.

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The ultrastructural changes of colitis (n = 10 per group, 2 μm). Ultrastructural changes in colon were examined with a transmission electron microscope. (A) CON, (B) DSS group, (C) FA+DSS group, (D) Pdn+DSS group, (E) mitochondrial damage score. The red arrow indicated intercellular junctions, and the blue arrow indicated the endoplasmic reticulum. N, the nucleus. All data are expressed as the mean ± SD. **p < 0.01, *p < 0.05, and ns p > 0.05. SD was shown on the top of each column.

4. Discussion

In this study, preventive FA supplementation increased the body weight, the SOD, GSH-Px, CAT, T-AOC activity, IL-2 and IL-10 contents, reduced the IL-6, IL-1β, FC, FL, and the MDA content, NLRP3 protein expression in the DSS-induced UC mice. FA attenuated the extensive inflammation infiltration, crypt destruction and the mitochondrial damage in the DSS-induced UC mice. Meanwhile, FA can effectively improve the intestinal flora in mice. All test results consistently showed that, compared to the DSS group, both the FA and the Pdn prevention group exhibited significant and directionally similar improvements across all indicators, but there is no significance between these groups. FA has no side effects compared with Pdn. It indicates that preventive FA supplementation is helpful to alleviate the DSS-induced UC in mice.

DSS, a polyanionic derivative of dextran, was toxic to colonic epithelial cells and directly induced colon injury (Kiesler et al., 2015). DSS-induced colitis model has become one of the most widely UC mode (Xu et al., 2022; Eichele and Kharbanda, 2017). A concentration of 3% DSS can successfully establish the model within 5 to 10 consecutive days (Lu et al., 2022). In this experiment, DSS damaged colonic morphology, increased inflammatory cytokines, inhibited anti-inflammatory cytokines, antioxidant capacity, and disordered gut microbiota. It indicates that the UC mice model was successfully established. FA exerted anti-inflammatory effects through regulation of inflammatory cytokine levels and amelioration of oxidative stress (Liu et al., 2025). Huang et al. demonstrated that 80 mg/kg FA significantly inhibited the inflammation cytokines, enhanced intestinal antioxidant activity in the intestine (Huang et al., 2025). However, since the model-inducing drug used in this experiment was DSS acute modeling, the preventive intragastric administration days of 80 mg/kg were extended from 7 days to 10 days. Pdn was a synthetic glucocorticoid with anti-inflammatory, anti-allergic, and immunosuppressive properties (Bruscoli et al., 2021). A dose of 5 mg/kg prednisone was used as a positive drug control for the treatment of IBD (Mencarelli et al., 2016). This is the dosage for therapeutic use. In this experiment, Pdn was administered as a preventive drug, and the drug concentration was reduced to 4 mg/kg.

FC and FL are biomarkers for inflammation in UC (Chen et al., 2023). IL-2, IL-6 and IL-1β are pleiotropic pro-inflammatory cytokines and played a prominent role in the pathogenesis of IBD (Kaur et al., 2020; Mao et al., 2018). IL-10 is a well-defined anti-inflammatory cytokine (Li et al., 2024). These cytokines performed multiple functions at sites of inflammation (Li and Zhang, 2023). In this experiment, FA markedly reduced the elevation of FC, FL, IL-1β and IL-6, and promoted the increase of IL-2 and IL-10. Fecal Cp and Lf levels had almost equivalent abilities in detecting clinical remission and tended to be higher in UC patients (Yamakawa et al., 2024). The reduction of the FC and FL in the FA+DSS and Pdn+DSS group indicates that the inflammation of UC was alleviated. IL-1β induced the expression of proinflammatory cytokine, promoting the amplification of inflammatory response (Pirklbauer et al., 2021). The deregulation of IL-6 was associated with chronic inflammation (Uciechowski and Dempke, 2020). The reduction of the IL-1β and IL-6 in the FA+DSS and Pdn+DSS group indicates that the inflammation of UC was alleviated. The function of IL-10 is to inhibit the production of pro-inflammatory cytokines, suppressing the inflammatory response (Li et al., 2024). The increase of IL-10 in the FA+DSS and Pdn+DSS group indicates that the anti-inflammation activity of UC mice is promoted. It indicates that the inflammation of UC was attenuated. However, in this experiment, the inflammation cytokine, IL-2, was increased. The both pro-inflammatory and anti-inflammatory function helps IL-2 to regulate the immune balance (Chen et al., 2024; Phillips et al., 2020). The promotion of IL-2 is similar to Linghu’s experiment (Linghu et al., 2022). Considering the pivotal role of the NLRP3 inflammasome in inflammatory responses, these results implied that FA might exert its anti-inflammatory effects. This study provides evidence that FA attenuates NLRP3-mediated inflammation. Ekhtiar et al. (2025) verified that FA reduced the expression of inflammatory factors, which provided theoretical basis for this experiment. FA attenuated the extensive inflammation infiltration, crypt destruction and the mitochondrial damage in the DSS-induced UC mice. The recover of the colitis morphology contributes to the inhibition of inflammation cytokines. Pdn elicited anti-inflammatory and immunosuppressive effects via pleiotropic modulation of immune function (Babbar et al., 2021). In this experiment, FA has the same protective effect on the inflammation with Pdn, but there is no significant difference between the two groups. Pdn has side effect in the treatment of UC. It indicates that FA inhibited the inflammation cytokine and promoted the anti-inflammation cytokines.

Natural antioxidant compounds exhibit ROS scavenging and increase antioxidant defense capacity to inhibit pro-oxidative enzymes, which may be useful in IBD treatment (Sahoo et al., 2023). In this experiment, the elevation of SOD, GSH-Px, and CAT activity induced by FA and Pdn indicated that FA restored the anti-oxidative function of UC mice. Another evidence showed that FA attenuated oxidative stress-induced cardiomyocyte injury (Sun et al., 2021). It supports our findings of FA on the antioxidative activity in the UC. Reactive oxygen species (ROS) and reactive nitrogen species (RNS), are produced at abnormally high levels in UC. Their destructive effects may contribute to the disease’s initiation and propagation. MDA can be measured in the blood and stool of patients with IBD (Muro et al., 2024). The reduction of MDA further confirmed FA and Pdn’s ability to mitigate oxidative damage in UC mice. T-AOC was a comprehensive indicator reflecting the overall antioxidant capacity (Xie et al., 2025). In this experiment, FA and Pdn restored T-AOC activity in UC mice. FA attenuated extensive inflammation infiltration, crypt destruction and mitochondria damage in colon of this experiment. The recover of the extensive inflammation infiltration, crypt destruction and the mitochondrial damage in the colitis morphology contributes to the increase of antioxidative function. In the gastrointestinal tract, infections enhanced ROS generation through pro-inflammatory cytokine upregulation (Sahoo et al., 2023). In this experiment, the reduction of the pro-inflammatory cytokine and the anti- inflammatory cytokine contribute to the inhibition of the oxidative stress in UC mice. It indicates that FA and Pdn promotes the antioxidative function of UC mice.

UC diminished intestinal microbiota diversity, depleting beneficial and expanding pathogenic bacteria (Yu Z. et al., 2023). DSS administration disrupted the gut microbiota, precipitating dysbiosis (Xu et al., 2021). In this experiment, the reduction of the pro-inflammatory cytokine also contributes to the recovery of the gut microbiota. Similarly, Clostridia, a component of the gut microbiota, is often associated with gut health in many studies. It was observed that the FA+DSS and Pdn+DSS group exhibited a significant reduction in bacteria of the class Clostridia. Gut microbiota dysbiosis triggered inflammation (Clinton and Cross, 2023). This increase promotes its ability to suppress inflammation.

Certain species within the genus Clostridium were important intestinal pathogens, and the reduction in their colonies could directly attenuate enteritis. Bacteroides, as one of the core members of the gut microbiota, played a crucial role in suppressing UC. Its mechanisms of action involve maintenance of microbial ecological balance, and anti-inflammatory effects mediated by metabolites. DSS-induced microbial disorder contributes to intestinal inflammation (Yang and Merlin, 2024). In the FA+DSS and Pdn+DSS group, a notable increase in Bacteroides was detected. Bacillus, also known as the spore-forming genus, and demonstrated multifaceted beneficial effects in suppressing IBD. Mechanistically, it acted by modulating gut microbiot. FA significantly changed gut microbiota composition in zebrafish (Yin et al., 2022). In the FA+DSS group of mice, a significant increase in the abundance of bacillus was observed. These findings collectively indicated that FA modulated the gut microbiota. Muribaculaceae plays essential roles in maintaining intestinal barrier integrity, modulating immune responses, and regulating energy metabolis, and was significantly depleted in the FA+DSS and Pdn+DSS group, suggesting that FA exposure under DSS-induced colitis may perturb microbial community homeostasis and contribute to dysbiosis. Conversely, a notable increase was observed in the relative abundance of beneficial genera such as Lactobacilli and Bifidobacteria. It indicates that they may be closely associated with the recovery pathological process in the FA+DSS and Pdn+DSS group. Additionally, FA increases the Bacteroides population, decreased dysbiosis and reduce multiple harmful consequences. A significant increase in Bacteroides was indeed observed in the FA+DSS and Pdn+DSS group. FA attenuated extensive inflammation infiltration, crypt destruction and mitochondria damage in colon of this experiment. Therefore, it can be concluded that FA can effectively improve the intestinal flora disorder in mice.

Therefore, FA and Pdn attenuate the UC through recover of the colon histopathology, reduction of inflammation, promotion of antioxidative function, regulation of intestinal flora. This provides a solid experimental foundation for its development as a novel and safe preventive agent for UC. The difference between the FA and Pdn group will be conducted in the future.

Funding Statement

The author(s) declared that financial support was received for this work and/or its publication. This research was funded by the grant from Postdoctoral Excellence Funding Program of Jilin Province.

Footnotes

Edited by: Huachun Pan, Longyan University, China

Reviewed by: Cong Zhang, Henan Agricultural University, China

Yuanyuan Zhang, Northeast Forestry University, China

Data availability statement

The data presented in the study are deposited in the the NCBI SRA repository, accession number PRJNA1445482.

Ethics statement

The animal study was approved by the IACUC of the Jilin Agriculture Science and Technology University. The study was conducted in accordance with the local legislation and institutional requirements.

Author contributions

ZZ: Conceptualization, Data curation, Investigation, Methodology, Writing – original draft, Writing – review & editing, Validation. ZD: Investigation, Writing – review & editing, Methodology, Supervision. HZ: Data curation, Software, Validation, Writing – review & editing. YZ: Conceptualization, Data curation, Investigation, Methodology, Software, Supervision, Writing – original draft, Writing – review & editing. CH: Supervision, Writing – review & editing, Resources, Validation, Visualization. WL: Supervision, Writing – review & editing, Conceptualization, Software. BY: Formal analysis, Project administration, Supervision, Validation, Writing – review & editing.

Conflict of interest

WL was employed by Jilin Zhengye Biological Products Ltd. Co.

The remaining author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Generative AI statement

The author(s) declared that Generative AI was not used in the creation of this manuscript.

Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.

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Supplementary material

The Supplementary material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2026.1767221/full#supplementary-material

Data_sheet_1.pdf (208.9KB, pdf)
Data_sheet_2.zip (3.2MB, zip)

References

  1. Ashique S., Mishra N., Garg A., Sibuh B. Z., Taneja P., Rai G., et al. (2023). Recent updates on correlation between reactive oxygen species and synbiotics for effective management of ulcerative colitis. Front. Nutr. 10:1126579. doi: 10.3389/fnut.2023.1126579, [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Babbar R., Dhiman S., Grover R., Kaur A., Arora S. (2021). A comprehensive review on therapeutic applications of Ferulic acid and its novel analogues: a brief literature. Mini Rev. Med. Chem. 21, 1578–1593. doi: 10.2174/1389557521666210120111702, [DOI] [PubMed] [Google Scholar]
  3. Bruscoli S., Febo M., Riccardi C., Migliorati G. (2021). Glucocorticoid therapy in inflammatory bowel disease: mechanisms and clinical practice. Front. Immunol. 12:691480. doi: 10.3389/fimmu.2021.691480, [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen Z., Balachandran Y. L., Chong W. P., Chan K. W. Y. (2024). Roles of cytokines in Alzheimer's disease. Int. J. Mol. Sci. 25:5803. doi: 10.3390/ijms25115803, [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen R., Li L., Tie Y., Chen M., Zhang S. (2023). Trajectory of fecal lactoferrin for predicting prognosis in ulcerative colitis. Precis. Clin. Med. 6:pbad022. doi: 10.1093/pcmedi/pbad022, [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clinton J. W., Cross R. K. (2023). Personalized treatment for Crohn's disease: current approaches and future directions. Clin. Exp. Gastroenterol. 16, 249–276. doi: 10.2147/ceg.S360248, [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Di Rienzo A., Marinelli L., Dimmito M. P., Toto E. C., Di Stefano A., Cacciatore I. (2024). Advancements in inflammatory bowel disease management: from traditional treatments to monoclonal antibodies and future drug delivery systems. Pharmaceutics 16:1185. doi: 10.3390/pharmaceutics16091185, [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eichele D. D., Kharbanda K. K. (2017). Dextran sodium sulfate colitis murine model: an indispensable tool for advancing our understanding of inflammatory bowel diseases pathogenesis. World J. Gastroenterol. 23, 6016–6029. doi: 10.3748/wjg.v23.i33.6016, [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ekhtiar M., Ghasemi-Dehnoo M., Azadegan-Dehkordi F., Bagheri N. (2025). Evaluation of anti-inflammatory and antioxidant effects of Ferulic acid and Quinic acid on acetic acid-induced ulcerative colitis in rats. J. Biochem. Mol. Toxicol. 39:e70169. doi: 10.1002/jbt.70169, [DOI] [PubMed] [Google Scholar]
  10. Ermis A., Aritici Colak G., Acikel-Elmas M., Arbak S., Kolgazi M. (2023). Ferulic acid treats gastric ulcer via suppressing oxidative stress and inflammation. Life 13:388. doi: 10.3390/life13020388, [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hu N., Liu J., Luo Y., Li Y. (2024). A comprehensive review of traditional Chinese medicine in treating neuropathic pain. Heliyon 10:e37350. doi: 10.1016/j.heliyon.2024.e37350, [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huang H., Li J., Zuo J., Qiu Y., Li B., Hu R., et al. (2025). Ferulic acid alleviates intestinal inflammatory damage in mice, associated with ameliorating intestinal barrier damage and gut microbiota. Animals 15:2698. doi: 10.3390/ani15182698, [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaur S., Bansal Y., Kumar R., Bansal G. (2020). A panoramic review of IL-6: structure, pathophysiological roles and inhibitors. Bioorg. Med. Chem. 28:115327. doi: 10.1016/j.bmc.2020.115327, [DOI] [PubMed] [Google Scholar]
  14. Kiesler P., Fuss I. J., Strober W. (2015). Experimental models of inflammatory bowel diseases. Cell. Mol. Gastroenterol. Hepatol. 1, 154–170. doi: 10.1016/j.jcmgh.2015.01.006, [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Li R., Ye J. J., Gan L., Zhang M., Sun D., Li Y., et al. (2024). Traumatic inflammatory response: pathophysiological role and clinical value of cytokines. Eur. J. Trauma Emerg. Surg. 50, 1313–1330. doi: 10.1007/s00068-023-02388-5, [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Li H., Zhang Q. (2023). Research Progress of flavonoids regulating endothelial function. Pharmaceuticals (Basel) 16:1201. doi: 10.3390/ph16091201, [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Linghu K. G., Ma Q., Xiong S. H., Zhao M., Chen Q., Xu W., et al. (2022). The "whole ingredients extract" of Astragali Radix improves the symptoms of dextran sulfate sodium-induced ulcerative colitis in mice through systemic immunomodulation. Chin. Med. 17:109. doi: 10.1186/s13020-022-00661-0, [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu J., Guan Y., Yang L., Fang H., Sun H., Sun Y., et al. (2025). Ferulic acid as an anti-inflammatory agent: insights into molecular mechanisms, pharmacokinetics and applications. Pharmaceuticals (Basel) 18:912. doi: 10.3390/ph18060912, [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Liu Y., Shi L., Qiu W., Shi Y. (2022). Ferulic acid exhibits anti-inflammatory effects by inducing autophagy and blocking NLRP3 inflammasome activation. Mol Cell Toxicol. 18, 509–519. doi: 10.1007/s13273-021-00219-5, [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lu C., Zhang X., Luo Y., Huang J., Yu M. (2022). Identification of CXCL10 and CXCL11 as the candidate genes involving the development of colitis-associated colorectal cancer. Front. Genet. 13:945414. doi: 10.3389/fgene.2022.945414, [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mao L., Kitani A., Strober W., Fuss I. J. (2018). The role of NLRP3 and IL-1β in the pathogenesis of inflammatory bowel disease. Front. Immunol. 9:2566. doi: 10.3389/fimmu.2018.02566, [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Martínková L., Grulich M., Pátek M., Křístková B., Winkler M. (2023). Bio-based valorization of lignin-derived phenolic compounds: a review. Biomolecules. 13:717. doi: 10.3390/biom13050717, [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mencarelli A., Cipriani S., Francisci D., Santucci L., Baldelli F., Distrutti E., et al. (2016). Highly specific blockade of CCR5 inhibits leukocyte trafficking and reduces mucosal inflammation in murine colitis. Sci. Rep. 6:30802. doi: 10.1038/srep30802, [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Muro P., Zhang L., Li S., Zhao Z., Jin T., Mao F., et al. (2024). The emerging role of oxidative stress in inflammatory bowel disease. Front. Endocrinol. 15:1390351. doi: 10.3389/fendo.2024.1390351, [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Phillips B. G., Wang Y., Ambati S., Ma P., Meagher R. B. (2020). Airways therapy of obstructive sleep apnea dramatically improves aberrant levels of soluble cytokines involved in autoimmune disease. Clin. Immunol. 221:108601. doi: 10.1016/j.clim.2020.108601, [DOI] [PubMed] [Google Scholar]
  26. Pirklbauer M., Sallaberger S., Staudinger P., Corazza U., Leierer J., Mayer G., et al. (2021). Empagliflozin inhibits IL-1β-mediated inflammatory response in human proximal tubular cells. Int. J. Mol. Sci. 22:5089. doi: 10.3390/ijms22105089, [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sahoo D. K., Heilmann R. M., Paital B., Patel A., Yadav V. K., Wong D., et al. (2023). Oxidative stress, hormones, and effects of natural antioxidants on intestinal inflammation in inflammatory bowel disease. Front. Endocrinol. 14:1217165. doi: 10.3389/fendo.2023.1217165, [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sun S., Ruan Y., Yan M., Xu K., Yang Y., Shen T., et al. (2021). Ferulic acid alleviates oxidative stress-induced cardiomyocyte injury by the regulation of miR-499-5p/p21 signal Cascade. Evid. Based Complement. Alternat. Med. 2021:1921457. doi: 10.1155/2021/1921457, [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Uciechowski P., Dempke W. C. M. (2020). Interleukin-6: a Masterplayer in the cytokine network. Oncology 98, 131–137. doi: 10.1159/000505099, [DOI] [PubMed] [Google Scholar]
  30. Wu Y., Ran L., Yang Y., Gao X., Peng M., Liu S., et al. (2023). Deferasirox alleviates DSS-induced ulcerative colitis in mice by inhibiting ferroptosis and improving intestinal microbiota. Life Sci. 314:121312. doi: 10.1016/j.lfs.2022.121312, [DOI] [PubMed] [Google Scholar]
  31. Xie Y., Gu Y., Li Z., Zhang L., Hei Y. (2025). Effects of exercise on different antioxidant enzymes and related indicators: a systematic review and meta-analysis of randomized controlled trials. Sci. Rep. 15:12518. doi: 10.1038/s41598-025-97101-4, [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Xu H. M., Huang H. L., Liu Y. D., Zhu J. Q., Zhou Y. L., Chen H. T., et al. (2021). Selection strategy of dextran sulfate sodium-induced acute or chronic colitis mouse models based on gut microbial profile. BMC Microbiol. 21:279. doi: 10.1186/s12866-021-02342-8, [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Xu Q., Sun W., Zhang J., Mei Y., Bao J., Hou S., et al. (2022). Inflammasome-targeting natural compounds in inflammatory bowel disease: mechanisms and therapeutic potential. Front. Immunol. 13:963291. doi: 10.3389/fimmu.2022.963291, [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yamakawa T., Miyake T., Yokoyama Y., Kazama T., Hayashi Y., Hirayama D., et al. (2024). Clinical performance of fecal calprotectin, lactoferrin, and hemoglobin for evaluating the disease activity of IBD and detecting colorectal tumors. JGH Open. 8:e13077. doi: 10.1002/jgh3.13077, [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamazaki M., Chung H., Xu Y., Qiu H. (2023). Trends in the prevalence and incidence of ulcerative colitis in Japan and the US. Int. J. Color. Dis. 38:135. doi: 10.1007/s00384-023-04417-6, [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yang C., Merlin D. (2024). Unveiling colitis: a journey through the dextran sodium sulfate-induced model. Inflamm. Bowel Dis. 30, 844–853. doi: 10.1093/ibd/izad312, [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yin X., Liu W., Chen H., Qi C., Chen H., Niu H., et al. (2022). Effects of ferulic acid on muscle development and intestinal microbiota of zebrafish. J. Anim. Physiol. Anim. Nutr. 106, 429–440. doi: 10.1111/jpn.13631, [DOI] [PubMed] [Google Scholar]
  38. Yu Z., Li D., Sun H. (2023). Herba Origani alleviated DSS-induced ulcerative colitis in mice through remolding gut microbiota to regulate bile acid and short-chain fatty acid metabolisms. Biomed. Pharmacother. 161:114409. doi: 10.1016/j.biopha.2023.114409, [DOI] [PubMed] [Google Scholar]
  39. Yu S., Qian H., Zhang D., Jiang Z. (2023). Ferulic acid relieved ulcerative colitis by inhibiting the TXNIP/NLRP3 pathway in rats. Cell Biol. Int. 47, 417–427. doi: 10.1002/cbin.11935, [DOI] [PubMed] [Google Scholar]
  40. Zhang D., Jing B., Chen Z. N., Li X., Shi H. M., Zheng Y. C., et al. (2023). Ferulic acid alleviates sciatica by inhibiting neuroinflammation and promoting nerve repair via the TLR4/NF-κB pathway. CNS Neurosci. Ther. 29, 1000–1011. doi: 10.1111/cns.14060, [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zhang D., Jing B., Chen Z., Li X., Shi H., Zheng Y., et al. (2022). Ferulic acid alleviates sciatica by inhibiting peripheral sensitization through the RhoA/p38MAPK signalling pathway. Phytomedicine 106:154420. doi: 10.1016/j.phymed.2022.154420, [DOI] [PubMed] [Google Scholar]
  42. Zhang W. W., Thakur K., Zhang J. G., Wei Z. J. (2024). Riboflavin ameliorates intestinal inflammation via immune modulation and alterations of gut microbiota homeostasis in DSS-colitis C57BL/6 mice. Food Funct. 15, 4109–4121. doi: 10.1039/d4fo00835a, [DOI] [PubMed] [Google Scholar]
  43. Zhang M., Yang D., Yu H., Li Q. (2021). MicroRNA-497 inhibits inflammation in DSS-induced IBD model mice and lipopolysaccharide-induced RAW264.7 cells via Wnt/β-catenin pathway. Int. Immunopharmacol. 101:108318. doi: 10.1016/j.intimp.2021.108318, [DOI] [PubMed] [Google Scholar]
  44. Zhu Y., Hu C., Zheng P., Miao L., Yan X., Li H., et al. (2016). Ginsenoside Rb1 alleviates aluminum chloride-induced rat osteoblasts dysfunction. Toxicology 368-369, 183–188. doi: 10.1016/j.tox.2016.07.014, [DOI] [PubMed] [Google Scholar]
  45. Zhu Y., Sun J., Yan M., Lian S., Hu B., Lv S., et al. (2020). The biological characteristics of the canine adenovirus type 1 from fox and the transcriptome analysis of the infected MDCK cell. Cell Biol. Int. 45, 936–947. doi: 10.1002/cbin.11537 [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Data_sheet_1.pdf (208.9KB, pdf)
Data_sheet_2.zip (3.2MB, zip)

Data Availability Statement

The data presented in the study are deposited in the the NCBI SRA repository, accession number PRJNA1445482.

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