Puccinia horiana for relieving ulcerative colitis and application thereof

Abstract

The application discloses a prillium fimicola for relieving ulcerative colitis and application thereof, and belongs to the technical field of microorganisms. The prillium fimicola N03 can improve body weight loss and disease activity index increase caused by ulcerative colitis, inhibit colon atrophy, protect the integrity of colon tissue structure, increase the number of goblet cells, reduce the inflammation level in the intestinal tract, can significantly up-regulate the expression of Nrf2 , Gpx2 and Nqo1 in the body of a mouse with colitis, effectively improve the impaired activation of Nrf2 signal pathway in the body of the mouse, and has a good relieving effect on colitis. The prillium fimicola N03 has a very wide application prospect when used for preparing a medicine composition for relieving ulcerative colitis and a fermented food.​​​​​​​​

Description

Technical Field

[0001] This invention relates to a strain of *Alternaria fenestrated* that relieves ulcerative colitis and its application, belonging to the field of microbial technology. Background Technology

[0002] Inflammatory bowel disease (IBD), primarily including ulcerative colitis (UC) and Crohn's disease (CD), is a group of chronic, relapsing inflammatory gastrointestinal diseases with unknown etiologies. Clinical manifestations include diarrhea, abdominal pain, and rectal bleeding, severely impacting patients' quality of life. In recent years, the incidence of IBD has been steadily increasing globally. Currently, clinical treatment for IBD mainly relies on aminosalicylic acid preparations, corticosteroids, immunosuppressants, and biologics. However, these drugs often suffer from significant individual variability in efficacy, marked side effects, easy development of drug resistance, and high relapse rates after discontinuation. Therefore, finding safe and effective new intervention strategies has become an urgent research need.

[0003] Gut microbiota dysbiosis is considered a core element in the development and progression of IBD. Studies have shown that IBD patients exhibit significantly reduced gut microbiota diversity, decreased abundance of beneficial bacteria, and a relative increase in potentially pathogenic bacteria. Among these, *Alistipes*, a strictly anaerobic functional genus within the Bacteroidetes phylum, exhibits complex correlations between its abundance in the human gut and host health status. Numerous epidemiological and clinical studies have reported that abnormal changes in *Alistipes* are associated with various diseases, including liver fibrosis, colorectal cancer, cardiovascular disease, mood disorders, and depression. Furthermore, a reduction in *Alistipes* is associated with IBD; in IBD patients, the abundance of *Alistipes* is significantly lower than in healthy controls.

[0004] Alistipes finegoldii It is an important member of the genus *Alistipes*, originally isolated from human appendix specimens. Current research suggests that... A. finegoldii It can ferment carbohydrates to produce short-chain fatty acids, such as propionic acid and succinic acid. These metabolites have been shown to have anti-inflammatory effects and maintain intestinal barrier function. This evidence suggests that... A.finegoldii It may be a symbiotic bacterium with anti-inflammatory potential. However, there is currently no information regarding specific... A.finegoldii There is a lack of research reports on the systematic functional validation of strains and their specific applications in alleviating ulcerative colitis, especially regarding their theoretical and practical basis for development as probiotic preparations. Therefore, altering the host gut metabolic microenvironment through microbial preparations is a promising intervention strategy with significant research value. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, this invention provides *Alternaria fentanil* and its application in the treatment of ulcerative colitis. This bacterium can activate the Nrf2 signaling pathway in colonic cells, thus exhibiting a good ameliorative effect on colitis.

[0006] This invention provides a strain of *Finniella fischeri* (… Alistipes finegoldii N03 was deposited on December 30, 2025, at the Institute of Microbiology, Guangdong Academy of Sciences, at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCC No: 67581.

[0007] The strain *Alternaria finniculum* was isolated from the colonic contents of DSS-induced colitis C57BL / 6 mice. Sequencing analysis revealed its 16S rDNA sequence to be shown in SEQ ID NO.1. The sequence was then compared with the nucleic acid sequence of *Alternaria finniculum* using NCBI, showing a nucleic acid sequence similarity exceeding 99%. Therefore, it was named *Alternaria finniculum* N03.

[0008] The *Alternaria fintelii* N03 described herein possesses the following properties: Cell characteristics: Gram-negative rod-shaped bacteria, non-spore-forming, non-flagellated, with a cell length of approximately 0.5–1.5 µm wide and 0.5–1.5 µm long. Colony characteristics: Forms distinct colonies on culture media, typically 1–2 mm in diameter, with a rounded shape on the upper surface, a raised center, regular edges, slightly whitish, opaque, and moist and smooth surface. Growth characteristics: This strain is a strict anaerobe, with an optimal growth temperature of 36–38℃ and an optimal growth pH of approximately 6.0–6.5. It grows well in glucose-containing media, and enters the late logarithmic growth phase or early stationary phase after 16–24 h of culture.

[0009] The present invention also provides a microbial inoculant containing the above-mentioned *Alternaria fintelii* NO3.

[0010] In one embodiment of the present invention, the number of *Alternaria fintelii* cells in the microbial agent is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0011] The present invention also provides a product containing the above-mentioned *Alternaria finniculum* NO3 or the above-mentioned microbial agent.

[0012] In one embodiment of the present invention, the viable count of *Alternaria fintelii* in the product is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0013] In one embodiment of the present invention, the product is food, medicine, or health product.

[0014] In one embodiment of the present invention, the food includes beverages, dairy products, or other foods containing the above-mentioned ingredients.

[0015] In one embodiment of the present invention, the pharmaceutical product comprises Alternaria fenestration NO3, a drug carrier, and / or pharmaceutical excipients.

[0016] In one embodiment of the present invention, the dosage form of the medicine or health product includes granules, capsules, tablets, pills or oral liquids.

[0017] In one embodiment of the present invention, the pharmaceutical excipient is a pharmaceutically acceptable excipient.

[0018] In one embodiment of the present invention, the acceptable excipients include one or more commonly used thickeners, antioxidants, pH adjusters, emulsifiers, preservatives, fillers, binders, wetting agents, disintegrants, lubricants, and flavoring agents.

[0019] In one embodiment of the present invention, the filler is starch, sucrose, lactose, calcium sulfate and / or microcrystalline cellulose.

[0020] In one embodiment of the present invention, the adhesive is a cellulose derivative, alginate, gelatin, and / or polyvinylpyrrolidone. In one embodiment of the present invention, the wetting agent is water, ethanol, starch and / or syrup. In one embodiment of the present invention, the disintegrant is sodium carboxymethyl starch, carboxypropyl cellulose, croscarmellose, agar, calcium carbonate and / or sodium bicarbonate.

[0021] In one embodiment of the present invention, the lubricant is talc, calcium stearate, magnesium stearate, micronized silica gel, and / or polyethylene glycol. In one embodiment of the present invention, the flavoring agent is a simple syrup, sucrose, lecithin, orange peel syrup, cherry syrup, lemon, fennel, peppermint oil, sodium alginate, gum arabic, gelatin, methylcellulose, sodium carboxymethylcellulose, citric acid, tartaric acid, and Cladosporium fenelii or sodium bicarbonate.

[0022] The present invention also provides the use of the above-mentioned *Alternaria fenestration*, or the above-mentioned microbial agent, in the preparation of a medicine for the prevention and / or treatment of ulcerative colitis.

[0023] In one embodiment of the present invention, the viable count of *Alternaria fintelii* in the product is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0024] In one embodiment of the present invention, the drug comprises *Alternaria fentanil*, a drug carrier, and / or pharmaceutical excipients.

[0025] In one embodiment of the present invention, the dosage form of the medicine includes granules, capsules, tablets, pills, or oral liquids.

[0026] In one embodiment of the present invention, the pharmaceutical excipient is a pharmaceutically acceptable excipient.

[0027] In one embodiment of the present invention, the acceptable excipients include one or more commonly used thickeners, antioxidants, pH adjusters, emulsifiers, preservatives, fillers, binders, wetting agents, disintegrants, lubricants, and flavoring agents.

[0028] In one embodiment of the present invention, the filler is starch, sucrose, lactose, calcium sulfate and / or microcrystalline cellulose.

[0029] In one embodiment of the present invention, the adhesive is a cellulose derivative, alginate, gelatin, and / or polyvinylpyrrolidone. In one embodiment of the present invention, the wetting agent is water, ethanol, starch and / or syrup. In one embodiment of the present invention, the disintegrant is sodium carboxymethyl starch, carboxypropyl cellulose, croscarmellose, agar, calcium carbonate and / or sodium bicarbonate.

[0030] In one embodiment of the present invention, the lubricant is talc, calcium stearate, magnesium stearate, micronized silica gel, and / or polyethylene glycol. In one embodiment of the present invention, the flavoring agent is a simple syrup, sucrose, lecithin, orange peel syrup, cherry syrup, lemon, fennel, peppermint oil, sodium alginate, gum arabic, gelatin, methylcellulose, sodium carboxymethylcellulose, citric acid, tartaric acid, and Cladosporium fenelii or sodium bicarbonate.

[0031] Beneficial effects The present invention, *Fernella fenestration* ( Alistipes finegoldii N03 is a strain of *Alternaria fentanil* screened from the intestines of colitis-affected mice, possessing a function in improving colitis. This strain also has adjunctive therapeutic effects for ulcerative colitis. Mouse experiments showed that this strain effectively counteracted weight loss, increased disease activity index, and colonic shortening caused by colitis, and effectively improved colonic epithelial cell damage, increased goblet cell number, and significantly reduced intestinal inflammation levels. These results provide strong theoretical support for the adjunctive treatment of ulcerative colitis with probiotics.

[0032] Preservation of biological materials A strain of *Fernandezium fenestration* was deposited at the Institute of Microbiology, Guangdong Academy of Sciences on December 30, 2025, and its taxonomic name is: Alistipes finegoldii The accession number is GDMCC No: 67581, and the accession address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences Institute of Microbiology. Attached Figure Description

[0033] Figure 1 Changes in body weight and disease activity index in mice with colitis during intervention with *Alternaria fenestration* N03; where A represents the changes in body weight of mice in each group during the intervention period, and B represents the changes in disease activity index of mice during the intervention period.

[0034] Figure 2 Effects of *Alternaria fenestrated* N03 on colon length in mice with colitis and representative colon images; where A is a representative colon image of each group of mice, and B is the colon length.

[0035] Figure 3 Effects of *Alternaria fenestration* N03 on colonic pathological damage in colitis mice; where A is a representative section of colonic tissue stained with HE and AB-PAS, B is the number of colonic goblet cells, and C is the colonic pathological damage score.

[0036] Figure 4 Effects of *Alternaria fentanil* N03 intervention on colonic inflammation levels in mice with colitis; where A represents IL-10 level, B represents IL-1β level, C represents TNF-α level, and D represents IL-6 level.

[0037] Figure 5 : The effects of *Alternaria fentanil* N03 intervention on colitis mice Nrf2 The effect of related gene expression; where A is Nrf2 Gene expression levels; B is Gpx2 Gene expression levels; C is Nqo1 Gene expression levels.

[0038] "*" indicates a significant difference compared to the model group (*: P <0.05; **: P <0.01; ***: P <0.001; ****: P <0.0001); some data in the tables are expressed as mean values. Data analysis was performed using SPSS 24 with one-way ANOVA and Tukey's HSD post-hoc test. Detailed Implementation

[0039] The mice used in the following examples were purchased from Jiangsu Jicui Experimental Animal Co., Ltd., and housed in a constant temperature (22 ± 2℃) and constant humidity (55 ± 10%) SPF environment with a 12-hour light / dark cycle, while being provided with free access to standard food and water. Experiments began after one week of acclimatization. The DSS (molecular weight 36,000-50,000 Da) used in the following examples were purchased from MP Company, USA. The IL-10, TNF-α, IL-6, and IL-1β EILSA kits used in the following examples were purchased from Nanjing Fomax Biotechnology Co., Ltd.; the BCA protein concentration assay kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.; and the reverse transcription and qPCR kits were purchased from Takara Bio Inc., Japan. All culture medium components used in the following examples were purchased from Shanghai Yuanye Co., Ltd.

[0040] The following examples involve culture media: Preparation of enrichment medium (g / L): Potassium dihydrogen phosphate 1.0, sodium bicarbonate 3.0, potassium chloride 9.0, sodium chloride 9.0, anhydrous magnesium sulfate 1.2, potassium chloride dihydrate 0.2, ferrous sulfate heptahydrate 0.01, acid-hydrolyzed casein 1.4, tryptone 2.34, bacterial tryptone 2.34, yeast extract 2.1, cysteine ​​hydrochloride 1.6, bile salts 0.8, heme chloride 0.1, Tween 1.0, vitamin K1 0.01, β-glucan 5.0, dissolved in 1 L of distilled water, mixed thoroughly, and then the pH was adjusted to 6.2-6.9. After sterilization at 115-121℃ for 15-20 min, the enrichment medium was obtained.

[0041] MRS liquid culture medium (g / L): Peptone 10.0, beef extract 8.0, yeast extract 4.0, glucose 20.0, dipotassium hydrogen phosphate 2.0, diammonium hydrogen citrate 2.0, sodium acetate 5.0, magnesium sulfate 0.2, manganese sulfate 0.04, Tween 1.0, dissolved in 1 L of distilled water, and cysteine ​​hydrochloride 0.5-1 g / L was added. The mixture was thoroughly mixed, and the pH was adjusted to 6.6-7.0. After sterilization at 115-121℃ for 15-20 min, the MRS liquid culture medium was obtained.

[0042] Preparation of MRS solid medium: Add 1.5-2% agar to the MRS liquid medium. Mix well, then adjust the pH to 6.6-7.0, and sterilize at 115-121℃ for 15-20 min to obtain the MRS solid medium.

[0043] MRS selective medium (g / L): Peptone 10.0, beef extract 8.0, yeast extract 4.0, dipotassium hydrogen phosphate 2.0, diammonium hydrogen citrate 2.0, sodium acetate 5.0, magnesium sulfate 0.2, manganese sulfate 0.04, Tween 1.0, β-glucan 5, dissolved in 1L distilled water, and cysteine ​​hydrochloride 0.5-1 g / L was added. The mixture was thoroughly mixed, and the pH was adjusted to 6.6-7.0. After sterilization at 115-121℃ for 15-20 min, the liquid medium was obtained.

[0044] BHI liquid medium (g / L): 10.0g peptone, 17.5g ox heart extract, 5.0g sodium chloride, 2.0g glucose, and 2.5g disodium hydrogen phosphate are dissolved in 1L of distilled water. 0.5g cysteine ​​hydrochloride is added and mixed thoroughly. The pH is then adjusted to 7.2-7.6. The mixture is sterilized at 115-121℃ for 15-20 min to obtain the BHI liquid medium.

[0045] Preparation of BHI solid medium: Add 1.5-2% agar to BHI liquid medium. Mix well, then adjust the pH to 7.2-7.6, and sterilize at 115-121℃ for 15-20 min to obtain the BHI solid medium.

[0046] BHI selective medium (g / L): BHI solid medium was prepared by adding 5 mg / L of heme (sterilized by membrane filtration), 10 mg / L of vitamin K1, 7.5 mg / L of vancomycin, and 100 mg / L of kanamycin.

[0047] The detection methods involved in the following embodiments are as follows: Disease Activity Index (DAI) Evaluation: Starting from day 0 of DSS treatment, the degree of diarrhea and bloody stool in mice were monitored daily. Combined with changes in body weight, the daily DAI (average of three indicators) was calculated according to the scoring criteria in Table 1, and the DAI change curve of mice during the modeling period was plotted.

[0048] Table 1 Scoring criteria for various indicators of the mouse disease activity index

[0049] AB-PAS staining and goblet cell number analysis of colon sections: Mouse colon tissue was fixed in 4% paraformaldehyde solution for 24 h, followed by dehydration, paraffin embedding, and cutting into 3-4 μm thick paraffin sections. After baking at 60 ℃ for 3–5 h, the sections were air-dried on glass slides and stored at room temperature for later use. AB-PAS staining was performed according to the following steps: (1) Dewaxing paraffin sections to water: Dewax the sections in xylene I for 20 min, dewax in xylene II for 20 min, soak in anhydrous ethanol I for 5 min, soak in anhydrous ethanol II for 5 min, soak in 75% alcohol for 5 min, and finally rinse with tap water.

[0050] (2) Alcian blue staining: Immerse the sections in Alcian blue staining solution for 10–15 min, and rinse gently with running water.

[0051] (3) Periodic acid-Schiff staining: sequentially undergo deiodic acid oxidation treatment for 5–8 min, rinse with running water; react with Schiff reagent for 15–20 min, rinse with running water.

[0052] (4) Dehydration, clearing and mounting: Soak the sections in anhydrous ethanol I, anhydrous ethanol II and anhydrous ethanol III for 5 min each, then clear them in xylene I and xylene II for 5 min each, and finally mount them with neutral resin.

[0053] (5) Image acquisition and analysis: Images were acquired using a pathological slide scanner, and the number of goblet cells in the colon slides was statistically analyzed using Image J 6.0 software.

[0054] HE staining and pathological analysis of colon sections: Colon tissue was fixed by immersion in 4% paraformaldehyde solution for 24 h, then dehydrated and embedded in paraffin, and cut into 3-4 μm thick paraffin sections for HE staining according to the following steps.

[0055] (1) Dewaxing paraffin sections to water: Dewax the sections sequentially with xylene I for 20 min, xylene II for 20 min, soak in anhydrous ethanol I for 5 min, soak in anhydrous ethanol II for 5 min, soak in 75% alcohol for 5 min, and wash with tap water.

[0056] (2) Hematoxylin staining: Immerse the sections in hematoxylin staining solution for 3-5 min, wash with tap water, use blue solution to reverse blue staining, and rinse with running water.

[0057] (3) Eosin staining: Dehydrate the sections in 85% ethanol and 95% ethanol for 5 min in sequence, and then soak them in eosin staining solution for 5 min.

[0058] (4) Dehydration and mounting: Soak the sections in anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, xylene II for 5 min, and finally mount with neutral resin.

[0059] (5) Microscopic examination, image acquisition and analysis.

[0060] The pathological score of tissue damage was determined according to the scoring criteria in Table 2. Histological score = epithelial damage score + inflammation severity score + lesion depth score.

[0061] Table 2 Histological Damage Scoring Criteria

[0062] Determination of colon tissue-related cytokine levels: 30 mg of colon tissue was taken and mixed with sterile PBS and two 3 mm grinding beads at a ratio of 1:9 (w / v). The tissue was thoroughly homogenized using a homogenizer. The resulting homogenate was centrifuged at 10,000 rpm for 10 min at 4 ℃, and the supernatant was collected for subsequent assays. The levels of IL-10, TNF-α, IL-6, and IL-1β in mouse colon tissue were detected using the corresponding ELISA kits. Specific operating procedures and precautions were performed according to the manufacturer's instructions. Additionally, the total protein concentration of the samples was determined using a BCA protein concentration assay kit, and the content of the target protein per mg of total protein was calculated.

[0063] RNA extraction and gene expression determination: Total RNA was extracted from colon tissue using Trizol reagent, and reverse transcription was performed on the total RNA using a reverse transcription kit. Using the cDNA obtained from reverse transcription as a template, a PCR reaction system (20 μL) was prepared using the TB Green Premix Ex Taq II kit (TAKARA), consisting of 10 μL TB Green Premix Ex Taq II (Tli RNaseH Plus), 6 μL dd H2O, 0.8 μL upstream primer, 0.8 μL downstream primer, 0.4 μL LROX Reference Dye II, and 2 μL cDNA template. Primers are detailed in Table 3, with β-actin serving as the internal reference gene. Reaction conditions: Initial denaturation at 50℃ for 2 min, followed by 95℃ for 30 s; then 40 cycles of 95℃ for 20 s, 60℃ for 30 s, and 72℃ for 30 s; extension at 72℃ for 5 min to terminate the reaction. Based on the Ct values ​​of the target gene and the internal reference gene, a 2... -△△Ct The relative expression level of the target gene is calculated using this method.

[0064] Table 3 Target gene and primer sequences

[0065] Example 1: Isolation and identification of Alternaria fintelii N03 1. Accumulation of fecal bacteria Seven days after the establishment of the colitis model in mice, the colonic contents were collected and rapidly transferred to an anaerobic incubator. An appropriate amount of sterile PBS containing 0.1% L-cysteine ​​hydrochloride was added, and the mixture was vortexed and then passed through a 100 μm sterile cell sieve to obtain the bacterial culture. The bacterial culture was added to enrichment medium at 2% (v / v) and cultured under shaking at 37°C for 24 h in an anaerobic environment.

[0066] 2. Isolation and purification of Lactobacillus Spread the diluted bacterial culture onto MRS solid medium and incubate for 2-3 days for selective culture of Lactobacillus. Select plates with appropriate colony counts, pick single colonies with neat edges, slightly white, opaque, moist and smooth surfaces, and uniform morphology from the solid medium, and inoculate them into 5 mL of liquid MRS selective medium. Incubate at 37°C under anaerobic conditions for 24 h to obtain purified culture.

[0067] 3. Preservation and Identification of Microbial Strains The bacterial culture with the best viability from step 2 was used as an amplification template, and PCR amplification was performed using 16S universal primers (see Table 4). The PCR system is shown in Table 5, and amplification was carried out on a PCR instrument according to the following procedure: pre-denaturation at 95℃ for 5 min; followed by 29 cycles of 95℃ for 15 s, 60℃ for 15 s, and 72℃ for 45 s; extension at 72℃ for 5 min; and termination of the reaction by cooling to 4℃.

[0068] The amplified products were analyzed by 1% agarose gel electrophoresis and then subjected to first-generation sequencing. High-quality sequences were extracted from the sequencing data and submitted to NCBI for BLAST alignment to retrieve relevant sequence annotation information. The results showed that this sequence had over 99% homology with the 16S rDNA sequence of [unspecified organism]. This strain is now named *Alternaria fentanil* N03 and deposited at the Institute of Microbiology, Guangdong Academy of Sciences.

[0069] Table 4 Primer Names and Sequences

[0070] Table 5 PCR System

[0071] Example 2: Effects of *Alternaria finniculina* NO3 on body weight and disease activity index in mice with ulcerative colitis The specific steps are as follows: 1. Preparation of cryopreservation agents: (1) Cultivation method: In a sterile anaerobic environment, the strain of Alternaria finnifolia N03 was streaked on MRS solid medium and cultured under anaerobic conditions for 48 h. After single colonies were formed, single colonies were picked and inoculated into MRS liquid medium. The culture was carried out anaerobically at 37℃ for 16-24 h to reach the stationary phase. The OD value at this time was 1.0~1.4, and the seed liquid was prepared.

[0072] (2) Preparation of protective agent: Weigh 1 g / L cysteine ​​hydrochloride and 200-300 g / L glycerol according to the final concentration, dissolve them evenly in distilled water, and sterilize at 115-121℃ for 15-20 min.

[0073] (3) Preparation of cryoprotectant: After centrifuging the seed culture of *Alternaria fintelii* N03 cultured to the stable period in step (1) (8000 rpm, 10 min, 4℃), wash it 1-2 times with sterile phosphate buffer (pH 7.2), and then resuspend the bacterial culture with the protectant prepared in step (2) to obtain *Alternaria fintelii* N03 cryoprotectant, and store it at -80℃ for later use.

[0074] 2. Preparation of *Alternaria fintelii* NO3 suspension: (1) Activated strain: Inoculate a single strain of Alternaria fenestrate N03 into BHI liquid medium and culture it anaerobically at 37°C for 16-24 h to reach the stationary phase.

[0075] (2) Preparation and administration of bacterial suspension: The exponential growth phase culture was centrifuged at 4°C and 6000 × g for 10 minutes to collect the bacterial cells. After discarding the supernatant, the bacterial cells were resuspended in pre-cooled sterile phosphate buffer solution and washed twice to thoroughly remove any culture medium residue. Finally, the bacterial cells were resuspended to the predetermined concentration (1 × 10^9 CFU / mL) using the same buffer solution and stored on ice for gavage. The entire process was completed within 2 hours to ensure bacterial viability. The actual viable cell concentration of the final bacterial suspension was verified by plate count. The experimental mice were administered 200 μL of the above bacterial suspension (approximately 2 × 10^8 CFU of viable cells) via gavage once daily.

[0076] 3. Experimental methods: This invention uses a 3% (w / v) DSS aqueous solution to induce ulcerative colitis in mice. Twelve healthy male C57BL / 6J mice aged 6 weeks were randomly divided into two groups (n=6 per group): a DSS-induced colitis model group (referred to as the model group) and a Cephalomyces fingeri N03+DSS group (referred to as the inoculation group).

[0077] The experimental procedure is shown in Table 6. After a one-week adaptation period: Model group: Free access to purified water from day 0 to day 7, free access to 3% DSS solution from day 8 to day 14, and 0.1 mL sterile phosphate buffer by gavage daily from day 0 to day 14.

[0078] The group receiving oral administration of bacteria was given free access to purified water from day 0 to day 7, and free access to 3% DSS solution from day 8 to day 14. From day 0 to day 14, the group received 0.2 mL of Alternaria finniculum N03 suspension via gavage daily.

[0079] During DSS treatment, mouse weight, stool loosening, and stool bleeding were monitored at the same time every day.

[0080] Table 6 Experimental Procedure

[0081] 4. Experimental Results: The changes in body weight of mice in each group are as follows: Figure 1 As shown in Figure A. The results showed that the body weight of the model group mice decreased to 81.16% of their initial body weight on the last day of DSS treatment. By intervening with *Alternaria finniculum* NO3 in the mice, the body weight of the mice in the irrigation group recovered to 84.65% of their initial body weight on day 7 after DSS treatment, which significantly reduced the body weight loss in the mice. P <0.01).

[0082] Changes in disease activity index in each group of mice are as follows: Figure 1 As shown in Figure B. The results indicated that DSS treatment gradually increased the disease activity index to 3.5, which increased the degree of loose stools, bloody stools, and weight loss. Intervention with *Cephalomyces fintelseri* NO3 significantly reduced the disease activity index in mice; on day 7 after DSS treatment, the disease activity index in the bead-drenched group recovered to 2.50 (…). P <0.01).

[0083] The above results indicate that the *Alternaria fenestrated* NO3 of this invention can significantly improve the body weight and disease activity index-related colitis phenotype in mice treated with DSS.

[0084] Example 3: Effects of *Alternaria finniculina* N03 on colon length and pathology in mice with ulcerative colitis The specific steps are as follows: The specific experimental method was the same as in Example 2. After the intervention ended on day 14, mice were euthanized by cervical dislocation, and the colon and cecum tissues were quickly dissected and separated. The colon was photographed and its length was measured, and it was divided into three parts: one part was preserved in RNAwait solution, one part was fixed in 10% neutral formalin solution, and the other part was frozen at -80 ℃ for later use. At the same time, the contents of the colon and cecum were collected and frozen together at -80 ℃ for subsequent analysis.

[0085] DSS induces colitis and colonic atrophy in mice; therefore, colonic length is often used as one of the criteria for assessing the severity of colitis. Figure 2 As shown in A and B, the average colon length in the DSS group was 4.15 cm. Intervention with *Alternaria fintelseri* NO3 in mice significantly improved the reduction in colon length. P <0.001), with an average length of 5.30 cm.

[0086] HE staining and pathological analysis of colon tissue yielded representative section images and pathological scoring results for each group, as follows: Figure 3 As shown in A and C. Observations revealed that the colonic tissue structure of the model group mice was incomplete, with disordered or even absent epithelial cells, destroyed crypt structures, congestion and edema of the mucosa and submucosa, and extensive inflammatory infiltration in the lamina propria and submucosa, with a pathological tissue score of 9.5. Compared with the model group, *Alternaria fintelii* NO3 significantly improved the pathological morphology of the colonic tissue, repaired mucosal damage, and reduced inflammatory cell invasion, with a pathological tissue score of 6.17. P <0.01).

[0087] AB-PAS staining and goblet cell number analysis were performed on colon tissue. Representative sections and goblet cell count results for each group are shown below. Figure 3 As shown in A and 3B. The number of goblet cells in the DSS group was 138. Intervention with *Alternaria fintelii* NO3 in mice significantly increased the number of goblet cells. P <0.01), with an average length of 260 goblet cells.

[0088] The above results indicate that *Alternaria fenestration* NO3 can significantly improve colonic atrophy in colitis mice, protect the integrity of colonic tissue structure, and increase the number of goblet cells.

[0089] Example 4: Effect of Cephalomycosis fenestration NO3 on intestinal inflammation levels in mice with ulcerative colitis The specific steps are as follows: The specific experimental method was the same as in Example 2. After the intervention ended on day 14, the mice were euthanized. The colon tissue was preserved at -80°C for ELISA detection.

[0090] For evaluation A. finegoldii To investigate the regulatory effects of IL-10 on intestinal mucosal inflammation, we examined the levels of key cytokines in colonic tissue. IL-10, as an important anti-inflammatory cytokine, plays a central role in maintaining intestinal immune homeostasis. Figure 4 As shown in Figure A, compared with the colitis model group, via... A. finegoldii The expression level of IL-10 protein in the colon tissue of the intervened mice was significantly increased. P<0.01), an increase of 2.07 times. This result indicates that... A. finegoldii It can effectively enhance the host's anti-inflammatory immune response.

[0091] Simultaneously, we analyzed three key pro-inflammatory factors: TNF-α, IL-6, and IL-1β, which collectively drive the progression of intestinal inflammation by activating inflammatory cells and cascade reactions. Figure 4 As shown in B~D A. finegoldii The treatment significantly reduced the expression levels of these pro-inflammatory factors. Specifically, compared with the model group, the concentration of TNF-α in the colon tissue of the enema group decreased from 32.04 pg / mg protein to 18.11 pg / mg protein; the concentration of IL-6 decreased from 177.88 pg / mg protein to 137.62 pg / mg protein; and the concentration of IL-1β decreased significantly from 408.35 pg / mg protein to 202.96 pg / mg protein.

[0092] The above results indicate that the *Alternaria fenestrated* NO3 of the present invention can significantly downregulate the level of inflammation in colitis mice, reduce the level of pro-inflammatory cytokines, increase the level of anti-inflammatory cytokines, restore intestinal immune balance, and effectively control the occurrence and development of colitis.

[0093] Example 5: Effect of Cephalomycosis fenestration NO3 on Nrf2 pathway activation in the intestine of mice with ulcerative colitis The specific experimental method was the same as in Example 2. After the intervention ended on day 14, the mice were euthanized. The colon tissue was rinsed with pre-cooled PBS, and 1 / 3 of it was soaked in RNA wait solution overnight and then frozen at -80°C for RNA extraction and qPCR experiments.

[0094] qPCR was used to detect intestinal contents in mice Nrf2 and its downstream key genes Gpx2 and Nqo1 The expression, the result is as follows Figure 5 As shown. Intervention with *Alternaria fintelii* NO3 in mice resulted in increased levels of *Alternaria fintelii* in the mouse intestines. Nrf2, Gpx2 and Nqo1 The expression of these components was significantly higher than that in the model group. Nrf2, Gpx2 and Nqo1 The expression levels were 1.34, 1.97, and 1.82 times higher than those in the model group, respectively.

[0095] The above results indicate that *Alternaria fenestration* NO3 of the present invention can significantly upregulate the levels of *Alternaria fenestration* in mice with colitis. Nrf2、 The expression of Gpx2 and Nqo1 effectively improved the expression of Gpx2 and Nqo1 in mice. Nrf2 Impaired activation of signaling pathways.

[0096] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A strain of *Fernandezium fentanyl* ( Alistipes finegoldii N03 was deposited on December 30, 2025, at the Institute of Microbiology, Guangdong Academy of Sciences, at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCCNo: 67581.

2. A microbial inoculant, characterized in that, Contains the *Alternaria finniculina* NO3 as described in claim 1.

3. The microbial agent as described in claim 2, characterized in that, In the aforementioned microbial inoculant, the viable count of *Alternaria finnicota* is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

4. A product characterized in that, The product contains *Alternaria finniculum* NO3 as described in claim 1.

5. The product as described in claim 4, characterized in that, In the product, the viable count of *Alternaria finniculina* NO3 is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

6. The product as described in claim 4 or 5, characterized in that, The product in question is food, medicine, or health supplement.

7. The product as described in claim 6, characterized in that, The food includes beverages, dairy products, or other foods containing *Alternaria fentanil* NO3 as described in claim 1.

8. The product as described in claim 6, characterized in that, The drug contains *Alternaria fentanil* NO3, and also contains a drug carrier and / or pharmaceutical excipients.

9. The use of the *Alternaria fenestration* NO3 as described in claim 1, or the microbial agent as described in claim 2 or 3, in the preparation of a medicine for the prevention and / or treatment of ulcerative colitis.

10. The product as described in claim 9, characterized in that, The dosage forms of the medicine include granules, capsules, tablets, pills, or oral liquids.

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