Application of triton in preparation of product for treating ulcerative colitis

Abstract

The invention belongs to the technical field of medicines, and discloses application of triton in preparation of food or medicines for preventing or treating colitis. Experimental research on a dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) mouse model shows that the Qula can remarkably slow down the weight loss rate of the UC mouse, reduce the disease activity index (DAI) and effectively improve pathological changes of colon tissue. In addition, the Qula can also regulate the imbalance of intestinal flora, and provides a new thought and basis for the development of ulcerative colitis treatment products.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to the application of Qula in the preparation of products for treating ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is a chronic, recurrent inflammatory bowel disease, which is mainly manifested as chronic inflammation of the colon and rectum. Clinical symptoms include abdominal pain, diarrhea, mucus or bloody stools, etc. In severe cases, it may lead to colon perforation, bleeding, and even cancer. It is common in adults aged 30 to 40 years old, and severe cases can cause disability. About 15% of UC patients need colectomy. According to relevant statistics, the annual direct or indirect cost of treating UC in Europe and the United States is 1.25 billion to 2.91 billion euros and 810 million to 14.9 billion US dollars, respectively. However, the etiology and pathogenesis of ulcerative colitis are not very clear. It is generally believed that the etiology of UC may be related to environmental factors, genetic factors, immune factors, etc.; among them, genetic and immune factors play a vital role in the pathogenesis of UC. Existing treatment methods mainly rely on anti-inflammatory drugs, immunosuppressants, antibiotics and biological agents, but these treatments are often accompanied by varying degrees of side effects and it is difficult to fundamentally cure the disease. Therefore, seeking new, safe and effective treatment options has become the focus of current research.

[0003] Qula is a traditional fermented dairy product made by lactic acid bacteria fermentation. It is rich in probiotics, lactic acid and bioactive substances, and has multiple physiological activities such as regulating intestinal microecology and enhancing immune function. In recent years, with the in-depth study of the mechanism of action of probiotics, Qula, as one of the sources of probiotics, has gradually been recognized as a potential functional food that can improve intestinal health through multiple pathways such as regulating the balance of intestinal flora and improving intestinal barrier function. However, the application of traditional Qula is mainly concentrated in the development of deep-processing products and strain screening. At present, there are no reports on the preventive and therapeutic effects of Qula on inflammatory bowel diseases such as UC. Summary of the invention

[0004] The object of the present invention is to provide an application of Qula in preparing food or medicine for preventing or treating colitis.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] Application of Qula in preparing food or medicine for preventing or treating colitis.

[0007] Preferably, the food or medicine contains Qula as an active ingredient.

[0008] Preferably, the prevention or treatment of colitis manifestations includes inhibiting colon shortening, reducing disease activity index, improving colon tissue pathological changes and regulating intestinal flora imbalance.

[0009] Preferably, the colitis is acute or chronic ulcerative colitis.

[0010] The present invention also provides a health food composition or a pharmaceutical composition for treating UC, which comprises the above-mentioned Qulazine and food or pharmaceutically acceptable adjuvants.

[0011] According to an embodiment of the present invention, the dosage of Qulaq in the food or medicine is 2.5-10 g / kg for mice.

[0012] According to an embodiment of the present invention, the food or medicine is an oral preparation.

[0013] Preferably, the oral preparation includes oral solution, tablet or capsule.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention has discovered a new use of Qula. The results of animal experiments show that supplementing Qula with an addition amount of 2.5% (w / w) for 2 weeks can significantly alleviate the weight loss of DSS-induced colitis mice, significantly resist the infiltration and erosion of inflammatory cells in the colon tissue of mice, significantly alleviate the shortening of the colon, blood in the stool and diarrhea in mice, and maintain the balance of the intestinal flora in mice. Thereby achieving an effective therapeutic effect, and providing a new idea for the treatment of UC, and also providing a new use of Qula for the treatment of UC. Compared with the current clinical drugs for the treatment of UC, Qula has high safety and low cost. The present invention searches for a completely new drug target from a different perspective, which not only provides a new therapeutic drug for the treatment of ulcerative colitis, but also broadens the drug use of Qula. At the same time, it also shows that Qula has development potential in the functional food market for the prevention of UC. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The effect of QuLa on the body weight of UC mice. * p<0.05.

[0017] Figure 2 The effect of QuLa on the body weight gain of UC mice. * p<0.05, ** p<0.01.

[0018] Figure 3 This is the effect of QuLa on DAI in UC mice. ** p<0.01, **** p<0.0001.

[0019] Figure 4 This is the effect of QuLa on the colon length of UC mice, specifically the pictures of the colon segments of the three groups.

[0020] Figure 5 The effect of QuLa on the colon intestinal mucosal injury index of UC mice. *** p<0.001, **** p<0.0001.

[0021] Figure 6 This is the effect of QuLa on the colon length value of UC mice, specifically the change in colon length value. * p<0.05, **** p<0.0001.

[0022] Figure 7 This shows the effect of QuLa on the pathological changes in the colon of UC mice, specifically the H&E staining images of pathological tissue sections of the three groups.

[0023] Figure 8 This is the effect of QuLa on the colon pathological score of UC mice, specifically the pathological score of colon tissue damage. *** p<0.001, **** p<0.0001.

[0024] Fig. 9 This is a β-diversity analysis of the intestinal flora of UC mice, specifically NMDS analysis. Fig. 9 , different coordinate positions represent the similarity of intestinal flora of each mouse.

[0025] Fig.10 This is the analysis of the abundance of the intestinal flora of UC mice by QuLa. Fig.10 , different color depths represent the abundance differences at the genus level of each intestinal flora.

[0026] Note: The present invention Figures 1 to 10 middle:

[0027] CON represents the control group, which was fed with AIN-93G growth diet;

[0028] M represents the model group, which was fed with AIN-93G growth feed;

[0029] QL represents the Qula intervention group, which was fed AIN-93G growing feed containing 2.5% (w / w) Qula. DETAILED DESCRIPTION

[0030] The present invention is further described below in conjunction with the embodiments of the present invention and the accompanying drawings, but the protection scope of the present invention is not limited thereto.

[0031] Example 1

[0032] Effects of QuLa on body weight and weight gain in UC mice

[0033] C57BL / 6J mice were housed under specific pathogen-free (SPF) conditions (ambient temperature 22±2℃, relative humidity 40-70%, 12h-12h light-dark cycle). At the beginning of the experiment, the mice were randomly divided into a control group, a model group, and an intervention group, with 8 mice in each group. The control group was given distilled water, and the other two groups were given free drinking of 1.5% DSS solution for 7 consecutive days to induce the ulcerative colitis model. During the first 7 days of modeling, all three groups were fed AIN93G feed. After the modeling was successful, the control group was given distilled water during the treatment period, and the other two groups were given free drinking of 1.5% DSS solution. At the same time, the control group and the model group were given AIN93G feed, and the intervention group was given AIN93G feed containing 2.5% Qula every day for 14 consecutive days. The experimental period was 3 weeks, during which all mice had free access to food and water, and the weight of the mice was recorded every day. The weight on day 0 was 100%, and the weight change on this basis was calculated in percentage.

[0034] according to Figure 1 , 2 As shown, compared with the model group, QuLa supplementation did not significantly improve the body weight of UC mice, but the body weight loss rate of UC mice after QuLa supplementation was significantly lower than that of the model group.

[0035] Example 2

[0036] Effects of QuLa on DAI in mice

[0037] The test drug dosage, mouse model construction, and administration method were the same as in Example 1. From the beginning of modeling to the end of the experiment, the status, weight change, eating conditions, stool characteristics, blood in stool, and overall status of each group of animals were continuously observed and recorded every day, and scored according to the following table to calculate the DAI score of each mouse. The calculation method is: DAI = (weight loss score + stool characteristics score + occult blood / blood in stool score + overall status score) Table 1 Disease Activity Index Scoring Table

[0038]

[0039]

[0040] The results are as follows Figure 3 As shown, at the end of the experiment, the DAI index of mice in the model group was significantly higher than that in the control group; the DAI of mice in the intervention group was significantly lower than that in the model group, indicating that Qula can reduce the DAI of UC mice and improve their fecal scores.

[0041] Example 3

[0042] Effects of QuLa on colon length and appearance of UC mice

[0043] 1. Take photos of colon tissue

[0044] The mice fed for 3 weeks in Example 1 were fasted for 6 hours (8:00-14:00), blood was collected from the eye sockets, and then the mice were killed by cervical dislocation. The colon tissues were removed and photographed.

[0045] like Figure 4 As shown in the results, DSS modeling can lead to intestinal inflammation and bleeding, which eventually leads to shortening of colon length. Qula can inhibit the shortening and bleeding of colon.

[0046] 2. Observation and statistics of colon appearance

[0047] The colon tissue was scored with the colon mucosa damage index (CMDI) according to Table 2.

[0048] Table 2 Intestinal mucosal injury index score table

[0049]

[0050]

[0051] The results are as follows Figure 5 As shown, the CMDI index of the model group was significantly higher than that of the CON group; compared with the model group, the CMDI index of the mice in the intervention group was significantly reduced, indicating that Qula has a protective effect on the colon of UC mice.

[0052] 3. Colon length statistics

[0053] The length of colon tissue was statistically analyzed using ImageJ (National Institute of Health, Bethesda, MD, USA) software.

[0054] like Figure 6 As shown in the figure, the model group showed a significant shortening of colon length, and Qula significantly inhibited the shortening of colon. There was a statistically significant difference between the intervention group and the model group.

[0055] Example 4

[0056] Effects of QuLa on pathological changes of colon in UC mice

[0057] 1. Colon tissue sections

[0058] The mouse colon tissue obtained in Example 3 was taken out from the formalin fixative, and pathological sections were prepared through the steps of dehydration, embedding, slicing, drying, dewaxing, hematoxylin-eosin staining (H&E) staining, etc. The sections were observed under a 5x and 18x microscope, and photos were taken.

[0059] according to Figure 7 As shown in the figure, compared with the control group, the colon tissue of the mice in the model group was swollen, the colon mucosal structure was severely damaged, a large number of crypts were atrophied and bifurcated, and a large number of inflammatory cells infiltrated the mucosal layer and submucosal layer; most of the crypt structures of the colon of the mice in the intervention group were relatively intact, some crypts were atrophied and damaged, and multiple inflammatory cells invading the mucosa to varying degrees were observed, but the degree of invasion of inflammatory cells was lower than that in the model group.

[0060] 2. Colon histopathological scoring

[0061] After taking pictures under a microscope, the histopathological scores of the H&E-stained colon tissues were calculated according to Table 3.

[0062] Table 3 Colon histopathology scoring table

[0063]

[0064]

[0065] according to Figure 8 As shown in the figure, the colon histopathological score of mice in the model group was much higher than that in the control group, and the colon histopathological score of mice in the intervention group was significantly lower than that in the model group, and the difference was statistically significant. This shows that QuLa can treat the damage of DSS to the colon of mice and reduce the severity of DSS-induced UC in mice.

[0066] Example 5

[0067] Effect of QuLa on β-diversity of intestinal flora in UC mice

[0068] The test drug dosage, mouse model construction, and administration method were the same as in Example 1. On the day the experiment ended, the mouse feces were collected, and based on the bacterial 16S V3-V4 variable region, the Illumina / Miseq second-generation sequencing technology platform was used for high-throughput sequencing technology and bioinformatics analysis methods to study the effect of Qula on the intestinal flora structure of DSS-induced colitis mice.

[0069] The β diversity results are as follows Fig. 9 As shown, NMDS analysis showed that DSS could significantly change the microbiota diversity of normal mice, causing it to deviate from the relative coordinates of the microbiota of normal mice. However, after intervention with Qula, the imbalance of the microbiota could be improved, causing the data coordinates of colitis mice to be biased towards the normal group.

[0070] Example 6

[0071] Effects of QuLa on the genus abundance of intestinal flora in UC mice

[0072] The test drug dosage, mouse model construction, and administration method were the same as those in Example 1. On the day the experiment ended, mouse feces were collected and 16S rRNA sequencing was performed to detect the abundance of intestinal flora at the genus level.

[0073] The results of intestinal flora abundance at the genus level are as follows Fig.10 As shown in the data, compared with the control group, the relative abundance of Bifidobacterium in the model group was significantly decreased, and the relative abundance of Escherichia-Shigella was significantly increased. After the intervention of Qula, the relative abundance changes of the above bacterial genera can be restored to the control group to a certain extent, indicating that Qula has a protective effect on the intestinal flora disorder of UC mice.

[0074] The above experimental results show that Qula can reduce the DAI of DSS-induced colitis mice, reduce the rate of weight loss and the degree of colon shortening. In addition, Qula can improve the imbalance of the flora of UC mice, proving the potential of Qula as a prebiotic in the treatment of UC.

[0075] The present invention is not limited to the above-mentioned embodiments. Without departing from the essential content of the present invention, any deformation, improvement and substitution that can be thought of by those skilled in the art shall fall within the scope of the present invention.

Claims

1. Application of Qula in preparing food or medicine for preventing or treating colitis.

2. The use according to claim 1, characterized in that: The food or medicine uses Qula as an active ingredient.

3. The use according to claim 1, characterized in that: The prevention or treatment of colitis symptoms includes inhibiting colon shortening, reducing disease activity index, improving colon tissue pathological changes and regulating intestinal flora imbalance.

4. The use according to claim 1, characterized in that: The colitis is acute or chronic ulcerative colitis.

5. The use according to claim 1, characterized in that: The dosage of QuLa for mice is: 1.0-4.0 g / kg.

6. The use according to claim 1, characterized in that: The food or medicine is a health food composition or a medicine composition.

7. The use according to claim 1, characterized in that: The food or medicine is an oral preparation.

8. The use according to claim 7, characterized in that: The oral preparations include oral liquid, tablets or capsules.

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