Use of a monkey ear ring preparation in the preparation of a drug for preventing and treating ulcerative colitis

Abstract

The application discloses application of a monkey ear ring preparation in preparation of a medicine for preventing and treating ulcerative colitis. The monkey ear ring is given in advance to relieve weight loss, shortening of colon length and epithelial damage, inflammatory cell infiltration and the like caused by dextran sulfate sodium (DSS) induced experimental colitis. Further through 16S rRNA and transcriptome sequencing, it is found that the monkey ear ring can effectively increase the abundance of beneficial bacteria and activate signal pathways related to T cell, NK cell (natural killer cell) and natural immune cell activation when treating DSS induced colitis.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical formulation technology, specifically relating to the application of monkey earring formulations in the preparation of drugs for the prevention and treatment of ulcerative colitis. Background Technology

[0002] Ulcerative colitis (UC) is a common chronic inflammatory bowel disease (IBD) characterized by inflammation of the colonic mucosa and submucosa, extending from the rectum to the colon, presenting with typical symptoms such as abdominal pain, bloody and mucous stools, and tenesmus. The pathogenesis of UC is complex, involving multiple immune responses, microbial dysbiosis, and genetic factors. Currently used medications for UC include antibiotics, glucocorticoids, and immunomodulators, but these drugs are not effective in curing the disease and long-term use can cause serious adverse reactions. Ulcerative colitis is a characteristic disease in Traditional Chinese Medicine (TCM), falling under the categories of "intestinal bleeding," "diarrhea," "chronic dysentery," and "bloody stools." Numerous clinical studies have shown that TCM may have unique advantages in treating chronic inflammatory diseases such as ulcerative colitis through its multiple active ingredients, multiple targets, and comprehensive conditioning.

[0003] Ulcerative colitis is often accompanied by an abnormal immune system response. The immune system of UC patients attacks their own colonic mucosal cells, leading to a persistent inflammatory response. This immune abnormality involves multiple inflammatory mediators (such as TNF-α, IL-6, and IL-1β) and may activate immune cells such as T cells and macrophages, thereby aggravating mucosal damage and ulcer formation (see Zhang Yujie, Liu Xingshan: Pathogenesis of Ulcerative Colitis and Progress in Traditional Chinese Medicine Intervention. Practical Journal of Traditional Chinese Internal Medicine. 2024, 38(06)). Chronic gastroenteritis (CGE), on the other hand, is usually caused by long-term gastrointestinal irritation (such as improper diet, drug stimulation, Helicobacter pylori infection, etc.), mainly manifested as mild to moderate inflammation of the gastrointestinal mucosa, usually without extensive involvement of the immune system. Its degree of inflammation is relatively mild and generally does not involve the activation of deep tissues and the systemic immune system. Therefore, the treatment of UC needs to more specifically regulate the immune system, especially the activity of inflammatory mediators and immune cells, while CGE focuses more on the protection of the gastrointestinal mucosa and local anti-inflammatory (see Ma Haifeng: Correlation Study between Ulcerative Gastroenteritis and Gastrointestinal Lesions. Modern Practical Medicine. 2020, 32(07)).

[0004] Current treatment for UC mainly involves drugs that modulate the inflammatory immune response, including aminosalicylic acid drugs, glucocorticoids, immunosuppressants, and biologics (Chen Xiaofen; Chen Yuhan; Ma Juan: (2023): Research progress on novel treatment methods for inflammatory bowel disease. Chinese Journal of General Practice. 2023, 26(27)). Treatment for CGE focuses more on the protection and regulation of the gastrointestinal tract, including antacids and proton pump inhibitors, gastrointestinal motility drugs, antibacterial drugs, and gastric mucosal protectants (Fang Jingyuan, Zou Tianhui: A re-understanding of chronic gastritis. Chinese Journal of Gastroenterology, 2021, 41(Z1)). CGE drugs, such as proton pump inhibitors and gastric mucosal protectants, have limited symptom relief effects on UC patients and cannot effectively inhibit the inflammatory response in the colon. These drugs cannot modulate the immune system, cannot target the pathological mechanism of UC, and may even delay the standardized treatment of UC. UC drugs, such as immunosuppressants and glucocorticoids, have many side effects and may worsen the gastrointestinal symptoms of patients with chronic gastroenteritis. For example, the use of glucocorticoids may lead to gastric ulcers and infection risks, which is not the best option for patients with non-immune-mediated CGE.

[0005] In conclusion, medications for treating ulcerative colitis and chronic gastroenteritis are not interchangeable due to differences in their pathological mechanisms, drug mechanisms of action, and clinical efficacy.

[0006] *Pithecellobium clypearia* Benth, a plant belonging to the genus *Pithecellobium* in the legume family, is mainly distributed in Guangdong, Zhejiang, and Fujian provinces, as well as Vietnam, Laos, and Myanmar. *Pithecellobium clypearia* was first recorded in the *Compendium of Materia Medica* by Li Shizhen in the Ming Dynasty, and is also mentioned in the *Lu Chuan Materia Medica* by Zhao Xuemin in the Qing Dynasty. Its dried stems and branches with leaves can be used medicinally. It has a slightly bitter and astringent taste, is slightly cold in nature, and enters the spleen, stomach, and liver meridians. It has the effects of clearing heat and detoxifying, cooling blood and reducing swelling, stopping diarrhea, and removing dampness and astringing sores. It is included in the *National Compendium of Chinese Herbal Medicine* and the *Guangdong Standards for Chinese Medicinal Materials*. The main chemical components of *Pithecellobium clypearia* include flavonoids, phenylpropanoids, organic phenolic acids, triterpenes, and steroids. Flavonoids are present in relatively high amounts, mainly including quercetin, luteolin, quercetin, and catechins. Monkey earring extract, made from monkey earrings, is the main ingredient in monkey earring anti-inflammatory tablets, which are mainly used to treat upper respiratory tract infections, acute pharyngitis, acute tonsillitis, and acute gastroenteritis.

[0007] There are currently no reports of using monkey earrings for the prevention and treatment of ulcerative colitis. Summary of the Invention

[0008] In view of the technical problem that existing technologies are difficult to cure chronic inflammatory bowel disease, especially ulcerative colitis, this invention provides the application of a monkey earring preparation in the preparation of drugs for the prevention and treatment of ulcerative colitis.

[0009] This invention is implemented as follows.

[0010] This invention provides the application of a monkey earring formulation in the preparation of drugs for the prevention and treatment of ulcerative colitis.

[0011] This invention also provides the application of monkey earring formulations in the preparation of drugs that alter RNA expression levels. The alteration of RNA expression levels includes enhancing at least one gene pathway related to the activation of T cells, NK cells, and innate immune cells, and reducing at least one gene pathway related to acute inflammatory responses and defense responses.

[0012] This invention provides the application of a monkey earring formulation in the preparation of a drug that alters the abundance of intestinal flora. The alteration of intestinal flora abundance includes reducing the abundance of harmful bacteria *Acinetobacter*, or increasing the abundance of at least one of beneficial bacteria *Akkermansia*, *Lactobacillus*, and *Bifidobacterium*.

[0013] Furthermore, the dosage form of the above-mentioned drugs is selected from one of the following: tablets, granules, suspensions, capsules, powders, emulsions, solutions, pellets, aerosols, gels, and drops.

[0014] This invention, through experiments, verifies that the monkey earring preparation applied to mice with ulcerative colitis can significantly alleviate intestinal inflammation, regulate gene pathways, and modulate the intestinal flora.

[0015] Specifically, compared with the prior art, the beneficial effects of the present invention are as follows:

[0016] 1. This invention explores the regulatory effect of monkey earring anti-inflammatory tablets on ulcerative colitis by inducing experimental ulcerative colitis with DSS. Experimental results show that monkey earring anti-inflammatory tablets can alleviate weight loss and disease aggravation during the modeling period, and also increase colon length. Further H&E staining revealed that epithelial damage was significantly reduced, inflammatory cell infiltration was significantly decreased, and histopathological scores were significantly reduced after administration of monkey earring anti-inflammatory tablets. This invention demonstrates that monkey earring anti-inflammatory tablets can effectively alleviate the disease progression of DSS-induced colitis.

[0017] 2. This invention explored the possible mechanism by which Monkey Earring Anti-inflammatory Tablets alleviate DSS-induced ulcerative colitis through transcriptome sequencing. GO analysis of 280 significantly upregulated genes and 239 downregulated genes revealed a significant increase in pathways related to the activation of T cells, NK cells (natural killer cells), and innate immune cells.

[0018] 3. This invention investigates the role of gut microbiota in alleviating DSS-induced ulcerative colitis with monkey earring anti-inflammatory tablets through 16S rRNA sequencing. Compared with the control group, the abundance of harmful bacteria Acinetobacter was significantly reduced after administration of monkey earring anti-inflammatory tablets, while the abundance of beneficial bacteria such as Akkermansia, Lactobacillus, and Bifidobacterium was increased. Attached Figure Description

[0019] The present invention and its beneficial technical effects will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0020] Figure 1 Experimental design for treating colitis with monkey earring anti-inflammatory tablet extract alone.

[0021] Figure 2 Figure showing the percentage change in body weight during the DSS modeling process in Experiment 1.

[0022] Figure 3 Disease activity index scoring chart during DSS modeling in Experiment 1.

[0023] Figure 4 Typical photographs of mouse colons and statistical charts of mouse colon lengths after DSS modeling in Experiment 1.

[0024] Figure 5 Typical H&E staining pattern of the terminal colon in Experiment 1 and its histopathological scoring statistics.

[0025] Figure 6 Heatmap of gene expression levels in colon tissue of colitis mice after RNA-seq detection in Experiment 1 and after administration of high-dose monkey earrings.

[0026] Figure 7 Volcano diagram of upregulated and downregulated genes in the colon of mice after high-dose monkey earring treatment for colitis in Experiment 1.

[0027] Figure 8 In Experiment 1, RNA-seq analysis revealed upregulated pathways enriched after high-dose earring treatment of colitis in monkeys.

[0028] Figure 9 In Experiment 1, RNA-seq analysis revealed downregulated pathways enriched after high-dose earring treatment of colitis in monkeys.

[0029] Figure 10 In Experiment 1, high-throughput metagenomic sequencing (16S rRNA-seq) was performed on the fecal bacteria genomes of the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg).

[0030] Figure 11 The results and statistical graphs of the microbial community structure of mouse fecal samples from the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg) in Experiment 1 are shown in the figure. The PCoA (PCo1, 44.53%; PCo2, 22.94%) analysis based on Bray-Curtis is used to analyze the microbial community structure of the mice.

[0031] Figure 12 In Experiment 1, UPGMA clustering tree analysis was performed on the microbial communities of the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg).

[0032] Figure 13 In Experiment 1, the relative abundance of microbial communities in the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg) was analyzed at the Order level.

[0033] Figure 14 In Experiment 1, the relative abundance of the microbial community in the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg) was analyzed at the Family level.

[0034] Figure 15 In Experiment 1, the relative abundance of microbial communities in the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg) was analyzed at the Genus level.

[0035] Figure 16 This is a flowchart of transcriptome sequencing data analysis.

[0036] Figure 17 This is a flowchart for 16S rRNA sequencing data analysis. Detailed Implementation

[0037] The following embodiments of the present invention are carried out under conventional conditions, and all commonly used chemical reagents used in the embodiments are commercially available products.

[0038] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0039] The terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, apparatus, product, or device that includes a series of steps is not limited to the steps or modules listed, but may optionally include steps not listed, or may optionally include other steps inherent to such process, method, product, or device.

[0040] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that these descriptions are merely exemplary and not intended to limit the scope of the invention. Furthermore, descriptions of well-known structures and technologies are omitted in the following description to avoid unnecessarily obscuring the concepts of the invention.

[0041] The following embodiments further describe the present invention, but these embodiments are not intended to limit the scope of protection of the present invention.

[0042] I. Experimental Design

[0043] The specific experimental steps of the animal experiment on the treatment of DSS-induced experimental ulcerative colitis with the monkey earring anti-inflammatory tablets of the present invention are as follows;

[0044] 1. Prepare mice: 20 male C57BL / 6J wild-type mice, weighing 25-30g, 8-10 weeks old;

[0045] 2. Prepare reagents: Sodium dextran sulfate (Cat#0216011080) was purchased from MP Biomedicals (USA).

[0046] Take 1250g of monkey earrings, add water and boil twice, the first time for 2 hours and the second time for 1 hour. Filter, combine the filtrates, concentrate into a thick paste, add an appropriate amount of starch and mix well, dry into an extract of about 200g, and you will get the monkey earring anti-inflammatory tablet extract.

[0047] 3. Grouping: The mice were randomly divided into groups of 5 mice each;

[0048] This invention demonstrates animal experiments used to verify the protective effect of monkey earring anti-inflammatory tablet extract against ulcerative colitis. The experimental groups are as follows:

[0049] Model group: physiological saline + DSS;

[0050] Low-dose group of monkey earrings: monkey earring anti-inflammatory tablet extract (1g / kg) + DSS;

[0051] High-dose group of monkey earrings: monkey earring anti-inflammatory tablet extract (2g / kg) + DSS;

[0052] 4. Allow the mice to adapt to the environment for one week to balance their gut microbiota.

[0053] 5. Modeling method: Mice were acclimatized for one week to balance the intestinal flora. After one week of pre-drug administration, they were given sterile DSS (2.5%) drinking water for 5 days, followed by sterile tap water for 3 days.

[0054] 6. Administration method: The monkey earring anti-inflammatory tablet extract was administered orally. After the intestinal flora of mice was balanced, they were pre-administered for one week, and then administered daily until the modeling was completed. Figure 1 ).

[0055] 7. Experimental steps:

[0056] 7.1 Weight changes and disease scoring: Weight was measured daily from the start of modeling until the end of modeling. Disease scoring was based on the shape of the mice's feces and the presence of blood in their stool during the modeling period.

[0057] score Stool characteristics degree of occult blood 0 Normal, not loose No occult blood or weak occult blood 1 Loose, but still formed Positive occult blood test 2 Loose, soft, barely formed The feces are dark brown. 3 Paste-like stool Bloody stool visible to the naked eye 4 Severe diarrhea Massive rectal bleeding

[0058] 7.2 Sample collection: Colons were collected to measure colon length. Approximately 2 cm of colon tissue was taken for transcriptome sequencing (RNA-seq), and 0.5-1 cm of the end of the colon was used for H&E staining. Feces from mice on days 5, 6, and 7 after DSS administration were collected for 16sRNA detection.

[0059] 7.3 Transcriptome Sequencing

[0060] 7.3.1 RNA Extraction

[0061] 1) Take an appropriate amount of tissue, grind it thoroughly in liquid nitrogen, then transfer it to a 1.5 mL centrifuge tube, add 1 mL of Trizol reagent, and mix thoroughly immediately;

[0062] 2) Let the mixed tissue stand at room temperature for 10 minutes to allow for complete lysis;

[0063] 3) Add 200 μL of chloroform, shake thoroughly to mix, centrifuge at 4°C and 12000 rpm for 10 min;

[0064] 4) Take the upper aqueous phase, add an equal volume of phenol:chloroform (25:24), mix thoroughly, centrifuge at 4℃ for 12000rpm for 10min;

[0065] 5) Take the upper aqueous phase, add an equal volume of chloroform, mix thoroughly, and centrifuge at 4℃ for 12000rpm for 10min;

[0066] 6) Take the upper aqueous phase, add an equal volume of isopropanol, let stand at -20℃ for 1 hour, centrifuge at 4℃, 12000rpm for 10min;

[0067] 7) Discard the supernatant, add 1 mL of 75% ethanol, wash the precipitate, centrifuge at 4°C for 8000 rpm for 5 min, and discard the supernatant;

[0068] 8) Repeat the previous step

[0069] 9) Briefly centrifuge, pipette to remove ethanol, and vacuum dry for 2-4 minutes;

[0070] 10) Add 20-50 μL of RNase-Free Water, dissolve at room temperature for 10 min, mix well, and then centrifuge briefly;

[0071] 11) Store at -80℃.

[0072] 7.3.2 RNA Quality Inspection

[0073] RNA quality testing mainly involves three methods: agarose gel electrophoresis, Nanodrop micro spectrophotometer detection, and Agilent 2100 detection.

[0074] 7.3.3 Transcriptome Library Construction

[0075] 1) mRNA purification and fragmentation

[0076] 2) cDNA first-strand synthesis

[0077] 3) cDNA second-strand synthesis

[0078] 4) Connector connection

[0079] 5) Purification of ligation products

[0080] 6) Library amplification

[0081] 7) Purification of amplification products using magnetic beads

[0082] 8) Document Quality Inspection

[0083] 7.3.4 Data Analysis

[0084] See Figure 16 Transcriptome sequencing data analysis flowchart.

[0085] 7.4 16S rRNA Sequencing Analysis

[0086] 7.4.1 DNA Extraction

[0087] 1) Transfer 150-200mg of fecal sample to a 2mL centrifuge tube, immediately add 1.2mL of Buffer SSL to the sample, and vortex for up to 1min to fully disperse the sample.

[0088] 2) Bathe in a 70℃ water bath for 10 minutes.

[0089] 3) Vortex for 15 seconds. Centrifuge at ≥14,000g for 10 minutes at room temperature. Transfer 250 μL of supernatant to a new 1.5 mL centrifuge tube.

[0090] 4) Add 20 μL of Proteinase K and 250 μL of Buffer AL supernatant. Invert and mix 10 times. Incubate at 70°C for 10 min.

[0091] 5) Add 250 μL of anhydrous ethanol to the sample and mix by inverting 10 times.

[0092] 6) Load HiPure DNA Mini Column I into a 2mL collection tube. Transfer the mixture to the column. Centrifuge at 10,000g for 30-60 seconds.

[0093] 7) Discard the eluent and reattach the column to the collection tube. Add 500 μL of Buffer GW1 to the column. Centrifuge at 10,000 g for 30-60 seconds.

[0094] 8) Discard the filtrate and reassemble the column into the collection tube. Add 650 μL of Buffer GW2 to the column.

[0095] 10,000g

[0096] 9) Discard the filtrate and collect the column. Centrifuge at 13,000g for 2 minutes to dry the column.

[0097] 10) Load the column into a 1.5 mL centrifuge tube. Add 50-200 μL of preheated Buffer AE (70 °C) to the center of the membrane in the column and incubate at room temperature for 2 min. Centrifuge at 13,000 g for 1 min.

[0098] 11) Discard the DNA binding column and store the DNA at -20°C.

[0099] 7.4.2 DNA Quality Inspection

[0100] 1) NanoDrop Micro-Spectrophotometer Detection: NanoDrop detection requires a sample volume of 2 μL, with a concentration measurement range of 2-3000 ng / μL. NanoDrop detects the OD value of nucleic acids to determine their purity. The A260 / A280 ratio should ideally be between 1.8 and 2.0. Another indicator of nucleic acid purity is the A260 / A230 ratio, which should be around 2.2. A value below 1.8 indicates significant contamination by organic matter such as sugars, peptides, phenols, and salt ions. This is likely due to the organic phase being aspirated during the aspiration of the upper aqueous phase.

[0101] 2) Agarose gel electrophoresis is used to detect the integrity of nucleic acid samples, whether degradation has occurred, and whether there is contamination such as protein. High-quality genomic DNA should appear as a single band after agarose gel electrophoresis.

[0102] 7.4.3 Polymerase Chain Reaction (PCR) Amplification

[0103] 7.4.4 Library Quantification and Sequencing

[0104] Library quality was assessed using an ABI StepOnePlus Real-Time PCR System (Life Technologies, USA), and sequencing was performed using Novaseq 6000 in PE250 mode pooling (NovaSeq6000S2Reagent Kit v1.5, Illumina, USA).

[0105] 7.4.5 Data Analysis

[0106] See Figure 17 Flowchart for 16S rRNA sequencing data analysis.

[0107] II. Experimental Results

[0108] 1. Monkey Earring Anti-inflammatory Tablets Significantly Alleviate Disease Progression in DSS-Induced Colitis

[0109] To investigate the protective effect of monkey earring anti-inflammatory tablets on ulcerative colitis, we used sodium dextran sulfate (DSS) to construct a mouse model of experimental colitis. The results showed that mice given monkey earring anti-inflammatory tablets (1 g / kg; 2 g / kg) beforehand exhibited less weight loss, a lower disease activity index (DAI), and a longer colon compared to control mice after DSS stimulation. Figure 2-4 (Tables 1-4). Further HE staining ( Figure 5 Table 5 shows that prior administration of monkey earring anti-inflammatory tablets (2g / kg) significantly reduced intestinal epithelial damage, decreased inflammatory cell infiltration, and significantly lowered histopathological scores. These preliminary results indicate that monkey earring anti-inflammatory tablets can effectively alleviate the progression of DSS-induced ulcerative colitis.

[0110] Table 1. Results of weight changes during DSS modeling.

[0111]

[0112]

[0113] Table 2 shows the percentage change in body weight during DSS modeling.

[0114]

[0115] Table 3. Disease Activity Index Scores During DSS Modeling

[0116]

[0117] Table 4 shows the colon length in mice after DSS modeling.

[0118]

[0119]

[0120] Table 5. Histopathological scores of the terminal colon

[0121]

[0122] 3. Monkey Earring Anti-inflammatory Tablets significantly alleviate intestinal inflammation and enhance intestinal T-cell immune responses.

[0123] To further explore the possible mechanism by which Monkey Earring Anti-inflammatory Tablets alleviate ulcerative colitis, we performed transcriptome sequencing on colon tissues from mice in the experimental group (2 g / kg) and control mice treated with Monkey Earring Anti-inflammatory Tablets (see heatmap and volcano plot). Figure 6-7 This indicates significant differences in RNA expression levels among different groups of mice. For example, the Ms4a4b, Gzmk, and Themis gene pathways were upregulated, while the IL10, Cxcl2, Cxcl3, and IL1a gene pathways were downregulated. Compared with the control group, mice given monkey earring anti-inflammatory tablets in advance showed significant downregulation of 239 genes and significant upregulation of 280 genes (log2FC > ±1; adjusted P < 0.05). GO analysis of the upregulated and downregulated genes (see...) Figure 8-9 Tables 6-7 show a significant decrease in genes related to "acute inflammatory response" and "defense response," while a significant increase was observed in pathways related to "T cell activation," "NK cell activation," and "innate immune cell activation." In summary, transcriptomic data indicate that Monkey Earring Anti-inflammatory Tablets can significantly alleviate intestinal inflammatory responses and antimicrobial peptide expression, and this process may depend on the activation of intestinal immune cells, especially T cells.

[0124] Table 6. Upregulated pathways enriched by RNA-seq analysis after high-dose monkey earring treatment for colitis.

[0125]

[0126]

[0127] Table 7. Downregulated pathways enriched by RNA-seq analysis after high-dose monkey earring treatment for colitis.

[0128]

[0129]

[0130] Table 8 shows the results of PcoA analysis of fecal samples from mice in the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg) based on Bray-Curtis (PCoA 44.93%; PCo2 22.54%).

[0131]

[0132] 4. Monkey Earring Anti-inflammatory Tablets alleviate the occurrence and development of DSS-induced colitis and regulate the intestinal flora.

[0133] To investigate whether the relief of experimental colitis by monkey earring anti-inflammatory tablets depends on the gut microbiota, we performed 16S rRNA sequencing on feces from mice in the control and monkey earring anti-inflammatory tablet administration groups. Shannon and Chao1 index calculations revealed that monkey earring anti-inflammatory tablets did not significantly regulate microbial diversity and community richness, while PCoA showed that the gut microbiota of mice in both the control and administration groups exhibited significant clustering. Figure 10-12 (Table 8). Further species composition analysis revealed that the Monkey Earring Anti-inflammatory Tablets significantly reduced the abundance of the harmful genus Acinetobacter and significantly increased the abundance of beneficial bacteria such as Akkermansia, Lactobacillus, and Bifidobacterium. Figure 13-15 (Tables 9-11). The results of this study indicate that Monkey Earring Anti-inflammatory Tablets can regulate the composition and distribution of intestinal flora, suggesting that Monkey Earring Anti-inflammatory Tablets regulate intestinal inflammation by altering intestinal immunity and intestinal microecology to regulate the intestinal mucosal barrier.

[0134] Table 9 shows the relative abundance of microbial communities at the Order level for the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg).

[0135]

[0136] Table 10 shows the relative abundance of microbial communities at the Family level in Experiment 1 for the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg).

[0137]

[0138]

[0139] Table 11 shows the relative abundance of microbial communities at the Genus level in Experiment 1 for the control group and the high-dose monkey earring anti-inflammatory tablet extract group (2g / kg).

[0140]

[0141] It should be noted that the specific features, structures, materials or characteristics described in this specification can be combined in any way. For the sake of brevity, not all possible combinations of the various technical features in the above embodiments have been described. Without contradiction, those skilled in the art can combine and integrate the different embodiments and features described in this specification.

[0142] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of a monkey earring formulation in the preparation of drugs for the prevention and treatment of ulcerative colitis.

2. Application of a monkey earring formulation in the preparation of drugs that alter RNA expression levels.

3. The application of the monkey earring formulation according to claim 2 in the preparation of drugs that alter RNA expression levels, characterized in that, The altered RNA expression levels include at least one gene pathway associated with the activation of T cells, NK cells, and innate immune cells, as well as at least one gene pathway associated with the reduction of acute inflammatory responses and defense responses.

4. The application of a monkey earring formulation in the preparation of drugs that alter the abundance of intestinal flora.

5. The application of the monkey earring formulation according to claim 4 in the preparation of drugs that alter the abundance of intestinal flora, characterized in that, The alteration of gut microbiota abundance includes reducing the abundance of harmful bacteria such as Acinetobacter spp., or increasing the abundance of at least one of beneficial bacteria such as Akkermansia spp., Lactobacillus spp., and Bifidobacterium spp.

6. The application according to any one of claims 1-5, characterized in that, The dosage form of the drug is selected from tablets, granules, suspensions, capsules, powders, emulsions, solutions, pellets, aerosols, gels, and drops.

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