Long-chain polythermia for treating irritable bowel syndrome and application of long-chain polythermia

Abstract

The invention discloses long-chain Dorenia for treating irritable bowel syndrome and application of the long-chain Dorenia, and belongs to the technical field of microorganisms. According to the present invention, the bacterial strain is Dorenia longiflora MW-023, and the preservation number of the bacterial strain is GDMCC No: 66941; experiments prove that the strain has the effect of improving the symptoms of the irritable bowel syndrome, can effectively improve weight loss caused by modeling, delay the first-time loose defecation time, reduce visceral sensitivity, inhibit inflammation, maintain intestinal barriers and regulate mood disorder, and has the effect of treating the irritable bowel syndrome.

Description

A Long-chain Dorperella for the Treatment of Irritable Bowel Syndrome and Its Application Technical Field

[0001] This invention relates to a long-chain Dorperella for treating irritable bowel syndrome and its application, belonging to the field of microbial technology. Background Technology

[0002] Irritable bowel syndrome (IBS) is a common chronic gastrointestinal disorder characterized by persistent or intermittent abdominal pain, changes in bowel habits, and may be accompanied by mood disorders such as anxiety. According to the Rome IV diagnostic criteria, IBS can be classified into four subtypes: diarrheal, constipation, mixed, and indeterminate, with the diarrheal subtype being the most common clinically. Epidemiological studies show that IBS has a high prevalence globally, reaching 5%–10% in most regions and approximately 4%–10% in Asian countries. This disease is often accompanied by mood disorders such as anxiety and depression, making its clinical management complex and a significant issue in the prevention and treatment of digestive system diseases. The pathogenesis of IBS is not fully understood, but current research suggests it involves multiple factors, including gut microbiota dysbiosis, visceral hypersensitivity, abnormal gastrointestinal motility, brain-gut axis dysregulation, intestinal mucosal immune activation, and impaired intestinal barrier function.

[0003] Currently, treatment for irritable bowel syndrome (IBS) mainly includes anti-inflammatory therapy, conventional drug therapy, and restoring intestinal permeability. Anti-inflammatory therapy can be divided into two categories: one primarily uses corticosteroids such as prednisolone; the other primarily uses antibiotics such as rifaximin. Secondly, conventional drug therapy, such as ramosetron and azimadolin, as well as major antidiarrheal drugs (such as montmorillonite powder and loperamide) and antispasmodics (such as trimebutine and pinaverium bromide), have shown positive effects; however, they are often accompanied by adverse reactions. For example, azimadolin may cause constipation in some patients; antidiarrheal drugs should not be used long-term to avoid masking the condition; and antispasmodics may cause symptoms such as dry mouth and dizziness. For the treatment of restoring intestinal permeability in IBS patients, probiotics and glutamine are commonly used. Probiotics normalize intestinal permeability through direct mechanisms such as stimulating the immune system, improving intestinal mucosal function, and altering the bacterial macroenvironment, or through indirect mechanisms such as regulating pathogenic invasion and bacterial adhesion. However, their value as a complementary therapy for IBS requires further evidence. In summary, existing drugs each have their own focus and shortcomings. Developing a drug that can simultaneously improve intestinal symptoms and emotional disorders with fewer side effects remains an important research direction.

[0004] Currently, there are no studies that clearly report the specific use of long-chain Dorperella lactis alone in the prevention or treatment of irritable bowel syndrome, and there is also a lack of systematic explanation of the development of related formulations and their mechanisms of action. Summary of the Invention

[0005] In view of this, the present invention aims to provide the use of *D. longan* in the preparation of pharmaceutical or probiotic formulations for the prevention or treatment of irritable bowel syndrome (IBS). This *D. longan* can alleviate IBS-related weight loss, delay the onset of first loose stools, regulate visceral hypersensitivity, improve intestinal inflammation, enhance intestinal barrier integrity, and reduce accompanying emotional symptoms, thereby exerting a comprehensive therapeutic effect. It also possesses advantages such as high safety and no side effects, providing a new and effective option for the microecological treatment of IBS.

[0006] This invention provides a strain of Dorealongicatena MW-023, which has been deposited at the Guangzhou Microbial Culture Collection Center, with accession number GDMCC No: 66941.

[0007] The present invention also provides a microbial preparation containing Dornicia longiformis MW-023.

[0008] In one embodiment, the microbial preparation contains at least 1 × 10⁻⁶ cells of *Dolphus longiformis* MW-023. 5 CFU / mL or 1×10 5 CFU / g.

[0009] The present invention also provides products containing the aforementioned Dorperella longiformis MW-023 or the aforementioned microbial preparation.

[0010] In one embodiment, the product includes a medicine or health product.

[0011] In one embodiment, the drug can improve weight loss caused by modeling.

[0012] In one embodiment, the drug can delay the time of the first loose stool.

[0013] In one embodiment, the drug can reduce the increase in visceral sensitivity caused by modeling.

[0014] In one embodiment, the drug is able to suppress inflammation.

[0015] In one embodiment, the drug is able to maintain the integrity of the intestinal barrier.

[0016] In one implementation, the drug can regulate mood disorders.

[0017] In one embodiment, the dosage form of the drug includes tablets, granules, capsules, pills, oral liquids, powders, lyophilized powders, drops, microcapsules, enteric-coated preparations, or immediate-release preparations.

[0018] In one embodiment, when the drug is in the form of an oral liquid, the content of *Dolphus longiformis* MW-023 in the drug is 1 × 10⁻⁶. 5 CFU / mL ~ 1×10 10 CFU / mL.

[0019] In one embodiment, the drug also includes a pharmaceutically acceptable carrier.

[0020] In one embodiment, the pharmaceutically acceptable carrier includes solvents, emulsifiers, disintegrants, fillers, binders, flavoring agents, colorants, lyophilization protectants, buffers, encapsulation materials, antioxidants, or prebiotics.

[0021] The present invention also provides the use of the aforementioned long-chain Dorperella MW-023 in the preparation of medicaments for the prevention and / or treatment of irritable bowel syndrome.

[0022] In one embodiment, the drug has at least one of the following functions: (1) improving weight loss; (2) delaying the time to first loose stool; (3) improving visceral sensitivity; (4) inhibiting inflammation; (5) maintaining intestinal barrier function; and (6) regulating mood disorders.

[0023] Beneficial effects of the present invention: The present invention relates to the application of *Dorcella longiformis* MW-023 in the preparation of drugs or probiotic preparations for the prevention and / or treatment of irritable bowel syndrome (IBS). Experiments show that this strain can significantly alleviate weight loss in model animals, delay the first loose stool passage, reduce visceral hypersensitivity, inhibit intestinal inflammatory response, enhance intestinal barrier integrity, and improve related mood disorders, thus exerting a comprehensive therapeutic effect on IBS. Compared with the commonly used therapeutic drug dexamethasone, *Dorcella longiformis* MW-023 shows superior improvement effects on multiple indicators.

[0024] Furthermore, *Dolphus longiformis* MW-023, isolated from the feces of healthy humans, belongs to the family Lachnospiraceae, a core symbiotic group of the human gut microbiota, and is generally considered non-pathogenic and non-allergenic. Therefore, the *Dolphus longiformis* MW-023 provided by this invention possesses high safety, no significant toxic side effects, and is suitable for long-term microecological intervention therapy for irritable bowel syndrome.

[0025] Biological material deposit: Dorealongicatena MW-023 (i.e., Dorealongicatena NSP014 in the deposit certificate), classified as Dorealongicatena, was deposited on September 9, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, located at: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences, Institute of Microbiology, Guangdong Province, deposit number: GDMCC No: 66941. Attached Figure Description

[0026] Figure 1 shows the changes in body weight during the experimental intervention; Figure 2 shows the time to the first loose stool during the experimental intervention; Figure 3 shows the mRNA expression level of visceral sensitivity factors; Figure 4 shows the level of inflammatory factors in colon tissue; Figure 5 shows the mRNA expression level of intestinal tight junction proteins; Figure 6 shows the trajectory and quantification of the open field experiment. Detailed Implementation

[0027] The YCFA liquid culture medium described in this invention is a commercially available product well-known in the art.

[0028] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0029] The Dorealongicatena NSP014 described in the following examples is the same as the Dorealongicatena NSP014 in the preservation certificate.

[0030] Example 1: Screening and Identification of Dolorella Longiformis MW-023 Dolorella Longiformis MW-023 was isolated from the feces of healthy human subjects. After obtaining fecal samples from healthy individuals, the samples were serially diluted and 10⁻⁶ samples were selected. -4 ~10 -8 The concentration was plated on YCFA solid medium and incubated at 37°C under anaerobic conditions for 24 hours. When single colonies were observed to grow, a single colony was picked and streaked onto YCFA solid medium. This process was repeated twice to complete purification and obtain single bacteria for identification. All related operations were performed in a strictly anaerobic glove box.

[0031] The selected *Dorealongicatena* MW-023 strain was inoculated onto YCFA solid medium. After the growth of round, white individual colonies, single colonies were picked and cultured in liquid medium. Sequence alignment using 16S rRNA identification confirmed it as *Dorealongicatena*, belonging to the Lachnospiraceae family. This family is a core symbiotic group in the human gut, possessing natural compatibility with the host's intestinal environment, lacking pathogenicity and allergenicity, and exhibiting high safety, making it suitable for use as a probiotic. *Dorealongicatena* MW-023 was deposited at the Guangdong Provincial Microbial Culture Collection Center, accession number GDMCC No: 66941.

[0032] Preparation of bacterial suspension: Take strain *Dolphobicella longiformis* MW-023 from cryopreservation tubes. When the strain is slightly thawed, pick an appropriate amount of bacteria using a sterile inoculation loop and streak it onto a YCFA agar plate for activation. Incubate at 37°C under anaerobic conditions for 48 hours. After activation, pick round, white single colonies and inoculate them into YCFA liquid medium. Incubate at 37°C under anaerobic conditions for 24 hours. Take the fermentation broth at the end of the logarithmic growth phase and centrifuge (4°C, 5000 rpm, 5 min) to collect the bacterial sludge. Wash the bacterial sludge with sterile, anaerobic PBS under the same centrifugation conditions, and then resuspend it in sterile, anaerobic PBS. In the above steps of fermentation culture and centrifugation resuspension, the fermentation volume and dilution factor for bacterial sludge resuspension were determined according to the growth curve of the strain to ensure that the viable cell concentration of the obtained bacterial suspension was 1×10⁻⁶. 10 CFU / mL, then take a portion and dilute it with PBS to 1×10⁻⁶. 8 CFU / mL.

[0033] Example 2: Therapeutic effect of *Dorcella longiformis* MW-023 on irritable bowel syndrome (IBS) 1. Modeling and Grouping Forty-eight 6-week-old (20.0 ± 1.0 g) SPF-grade C57BL / 6J male mice were randomly divided into four groups after one week of acclimatization: normal control group (Group B), model group (Group M), positive control group (Mesalazine), and *Dorcella longiformis* MW-023 intervention group (Group DL). Mice were allowed free access to food and sterilized drinking water for the first 7 days to acclimatize. Modeling began on day 8 (intervention time was counted as day 1). Mice in the model group were administered a 0.5 g / mL concentration of senna leaf decoction once daily via gavage at a dose of 10 mL / kg. The normal control group was administered the same amount of PBS via gavage. One hour after gavage, the mice in the model group were restrained for another hour using a centrifuge tube fixation method: 50 mL centrifuge tubes with a small opening at the tip were used; mice were placed inside the tubes, and the openings were plugged with foam blocks to restrict their free movement. After unrestraint, the mice were placed in cages lined with filter paper, and their defecation was observed for 6 hours. The bedding was replaced after 6 hours. During restraint modeling, the food was removed from each cage, and replaced after each day's restraint modeling. Model preparation and observation were performed at the same time each day to avoid interference from circadian rhythms. Modeling was conducted throughout the experimental period (8-22 days); the intervention period began on day 13 (day 6 of intervention), with the introduction of drugs and *D. longans* bacteria. After 6 hours of restraint each day, the positive control group received 300 mg mesalazine / kg of mice via gavage, while the intervention group received 0.2 mL of *D. longans* bacteria (MW-023) at a concentration of 1×10⁻⁶. 8 The mice were treated with CFU / mL of Dolorella vulgaris culture for 10 consecutive days. On day 23, all mice were euthanized by cervical dislocation, and biological samples were collected for subsequent research.

[0034] Table 1 Experimental Group Settings

[0035] 2. Detection methods and results (1) Long chain Dorsal bacteria MW-023 reduced the weight loss of mice with irritable bowel syndrome. The mice were weighed at a fixed time of 9:00 every morning, and the weight change during the 15-day intervention period was recorded.

[0036] As shown in Figure 1, compared with the model group, both the mesalazine group and the *D. longan* MW-023 group significantly slowed down the weight loss. Although there was no significant difference between the drug treatment group and the *D. longan* MW-023 group, the body weight of mice in the AC group continued to decrease after 14 days, indicating that the drug was difficult to maintain long-term effectiveness; while the *D. longan* MW-023 group showed a trend of stabilizing body weight in the later stage of the experiment, which is presumably due to the effective colonization and enrichment of the strain in vivo. Based on this, it can be considered that *D. longan* MW-023 may show better therapeutic effects in long-term treatment.

[0037] (2) During the intervention of Doxorubicin MW-023 to alleviate the abnormal defecation of mice with irritable bowel syndrome, the mice were placed in a mouse cage lined with filter paper after being untied every day, and the time of their first defecation of loose stool was recorded.

[0038] As shown in Figure 2, compared with the model group, both the mesalazine group and the *D. longan* MW-023 group delayed the time of the first loose stool in mice, although the differences among the three groups were not statistically significant. Considering that diarrhea-predominant irritable bowel syndrome is often accompanied by abnormal defecation and urge diarrhea, the above results indicate that *D. longan* MW-023 has a drug-like effect in regulating defecation, and based on its lack of side effects, this strain is more suitable as a treatment for irritable bowel syndrome.

[0039] (3) Long-chain Dorperella MW-023 reduces visceral sensitivity in mice with irritable bowel syndrome. Patients with irritable bowel syndrome often have abdominal discomfort symptoms such as abdominal pain. The mechanism of its occurrence is closely related to visceral hypersensitivity, which can amplify the transmission of intestinal pain signals. After the experiment, the prefrontal cortex of mice was collected, and the expression levels of visceral sensitivity factors CGRP and TRPV1 were detected.

[0040] As shown in Figure 3, both the mesalazine group and the *D. longan* MW-023 group significantly reduced the expression levels of visceral sensitivity factors in mice with irritable bowel syndrome (IBS), indicating that both have the potential to improve abdominal pain. Although the difference between the two groups did not reach statistical significance, the *D. longan* MW-023 group showed a better trend in reducing the expression of the above factors. The *D. longan* MW-023 group reduced the expression level of the visceral sensitivity factor CGRP from 1.535 in the model group to 0.891, which was better than the 0.942 in the drug group; the *D. longan* MW-023 group reduced the expression level of the visceral sensitivity factor TRPV1 from 1.130 in the model group to 0.627, which was better than the 0.754 in the drug group. Based on the positive effect of this strain in regulating visceral sensitivity and its lack of drug side effects, *D. longan* MW-023 has good application prospects in the treatment of IBS.

[0041] (4) Long-chain Dorperella MW-023 inhibits low-grade inflammation in mice with irritable bowel syndrome (IBS). IBS is often accompanied by low-grade intestinal inflammation. To evaluate the effects of different interventions on inflammation, the expression levels of inflammatory factors in the colon tissue of mice were detected after the experiment.

[0042] As shown in Figure 4, regarding pro-inflammatory factors, both the mesalazine group and the *D. longans* MW-023 group effectively reduced IL-6 expression, with comparable effects. However, in regulating the anti-inflammatory factor IL-10, the *D. longans* MW-023 group showed a significantly better effect than the mesalazine group. The mesalazine group only increased IL-10 expression from 113.851 pg / mg protein in the model group to 155.848 pg / mg protein, while *D. longans* MW-023 increased IL-10 expression to 220.54 pg / mg protein. In conclusion, *D. longans* MW-023 can synergistically regulate both pro- and anti-inflammatory aspects, possessing a more comprehensive function in improving intestinal inflammation and showing greater potential for application in the treatment of irritable bowel syndrome.

[0043] (5) Long-chain Dorperella MW-023 maintains the intestinal barrier integrity in irritable bowel syndrome (IBS) mice. IBS patients often suffer from impaired intestinal barrier function due to abnormal defecation symptoms such as diarrhea. To evaluate the effects of different interventions on intestinal barrier integrity, this invention detected the expression levels of intestinal barrier-related factors in mouse colon tissue after the experiment.

[0044] As shown in Figure 5, *Dolphus longiformis* MW-023 demonstrated superior performance compared to mesalazine in promoting intestinal barrier integrity. While mesalazine increased Claudin-1 expression (1.19), the difference was not significant compared to the model group (0.97), whereas *Dolphus longiformis* MW-023 significantly increased Claudin-1 expression (1.45) to a level similar to the control group (1.57). Although mesalazine increased Occludin expression, the increase was limited, rising from 1.068 in the model group to 1.618, while *Dolphus longiformis* MW-023 significantly increased Occludin expression to 2.074, closer to the control group (2.755). This effect may stem from its characteristics as a native gut microbiota, which can enhance synergistic effects among microbiota by regulating the gut microbiota, thereby more effectively maintaining barrier function. Therefore, Dorperella longiformis MW-023 has significant potential in repairing the intestinal barrier and is suitable for the treatment of irritable bowel syndrome.

[0045] (6) *Dorhizium anisopliae* MW-023 alleviates affective symptoms in mice with irritable bowel syndrome. An open field test (OFT) was conducted on day 14 of the intervention period (day 29 of the experiment). At the start of the experiment, mice were placed in the center of a white open field test chamber (40 cm × 40 cm × 60 cm), and their activities were recorded for 5 minutes using a camera. After each mouse's test, the open field test chamber was cleaned with 75% alcohol to remove mouse excrement and odor to avoid interference with subsequent mouse tests. Finally, the 5-minute behavioral data recorded for each mouse were analyzed, recording the mouse's movement trajectory and total distance traveled.

[0046] In open field experiments, the movement trajectories of mice contain key information about their behavior and emotional state. Total movement distance reflects the mice's basic motor and exploratory abilities, serving as a fundamental indicator for assessing their behavioral activity; while behavior across the central zone is considered a core behavioral indicator for evaluating anxiety-like behavior and mental stress. Combining motor ability with emotional expression in the analysis effectively eliminates the interference of simple activity changes on anxiety assessment, thus providing a reliable behavioral basis for a comprehensive and accurate interpretation of their overall mental state. The *D. longans* MW-023 intervention group showed superior improvement in movement trajectory compared to the mesalazine group, indicating a more significant effect in alleviating the mental and behavioral abnormalities associated with irritable bowel syndrome. Mesalazine, a conventional intestinal anti-inflammatory drug, primarily improves peripheral physiological symptoms, increasing movement distance from 1068.64 cm in the model group to 1685.01 cm, while *D. longans* MW-023 increased movement distance to 1901.91 cm, closer to the control group (2483.29 cm). Therefore, the combination of these long-chain Dorperella MW-023 shows a more comprehensive therapeutic potential in improving the mental and behavioral abnormalities associated with irritable bowel syndrome.

[0047] The above data show that when the long-chain Dorperella MW-023 of this invention is used alone to intervene in irritable bowel syndrome (IBS) model mice, the weight loss of IBS mice is slowed down, the visceral sensitivity is significantly reduced, the time to first loose stool is delayed, low-grade inflammation is suppressed, the intestinal barrier integrity is maintained, and the emotional disturbance of mice is reduced. It can effectively treat IBS when used alone.

[0048] 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 long-chain Dorea longicatena MW-023 has been deposited at the Guangzhou Provincial Microbial Culture Collection Center, with accession number GDMCC No: 66941.

2. A microbial preparation containing the long-chain Dorperella MW-023 as described in claim 1.

3. The microbial preparation as described in claim 2, characterized in that, In the aforementioned microbial preparation, the bacterial count of *Dolphobicia longiformis* MW-023 is not less than 1 × 10⁻⁶. 5 CFU / mL or 1×10 5 CFU / g.

4. A product containing the long-chain Dornerella vaginalis MW-023 of claim 1 or the microbial preparation of claim 2 or 3, characterized in that, The products mentioned include medicines or health supplements.

5. The product according to claim 4, characterized in that, The dosage forms of the drug include tablets, granules, capsules, pills, oral liquids, powders, lyophilized powders, drops, microcapsules, enteric-coated preparations, or immediate-release preparations.

6. The product according to claim 5, characterized in that, When the drug is in the form of an oral liquid, the content of *Dolphobicia longiformis* MW-023 in the drug is 1×10⁻⁶. 5 CFU / mL ~ 1×10 10 CFU / mL.

7. The product according to claim 6, characterized in that, The drug also includes a pharmaceutically acceptable carrier.

8. The product according to claim 7, characterized in that, Pharmaceutically acceptable carriers include solvents, emulsifiers, disintegrants, fillers, binders, flavoring agents, colorants, lyophilization protectants, buffers, encapsulation materials, antioxidants, or prebiotics.

9. The use of the long-chain Dorperella MW-023 of claim 1 in the preparation of a medicament for the prevention and / or treatment of irritable bowel syndrome.

10. The application as described in claim 9, characterized in that, The drug has at least one of the following functions: (1) improving weight loss; (2) delaying the time of first loose stool; (3) improving visceral sensitivity; (4) inhibiting inflammation; (5) maintaining intestinal barrier function; and (6) regulating mood disorders.

Images