CN114209810A

CN114209810A - Application of lycium barbarum glycopeptide in preparation of medicine for preventing or treating inflammatory bowel disease

Info

Publication number: CN114209810A
Application number: CN202210008718.XA
Authority: CN
Inventors: 秦蒙; 黄鲁; 黄翊淳; 郑颖慧; 苏国辉; 张力; 肖佳; 于哲雄; 王金霞; 樊福
Original assignee: Ningxia Qipeptide Technology Co ltd
Current assignee: Ningxia Qipeptide Technology Co ltd
Priority date: 2022-01-06
Filing date: 2022-01-06
Publication date: 2022-03-22
Anticipated expiration: 2042-01-06
Also published as: CN114209810B

Abstract

The invention belongs to the field of medicines, and particularly relates to application of lycium barbarum glycopeptide in preparation of a medicine for preventing or treating inflammatory bowel disease. The experimental result of the mouse acute ulcerative colitis model induced by Dextran Sodium Sulfate (DSS) shows that the lycium barbarum glycopeptide administration treatment can remarkably advance the chronic remission stage, remarkably relieve the symptoms of acute colitis, effectively reduce inflammatory reaction and show obvious anti-inflammatory effect. The experimental results show that the lycium barbarum glycopeptide serving as the traditional Chinese medicinal material extract product can be effectively used for preventing and treating inflammatory bowel diseases, has a prospect of developing a medicament for preventing and treating inflammatory bowel diseases, provides more treatment options for clinically preventing or treating inflammatory bowel diseases, and has important social benefits and economic values.

Description

Application of lycium barbarum glycopeptide in preparation of medicine for preventing or treating inflammatory bowel disease

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of lycium barbarum glycopeptide in preparation of a medicine for preventing or treating inflammatory bowel disease.

Background

Inflammatory Bowel Disease (IBD) including Crohn's Disease (CD) and Ulcerative Colitis (UC) is a chronic nonspecific inflammatory disease of the intestinal tract that involves the mucosa of the ileum, rectum and colon, with repeated courses of disease, clinically manifested by symptoms such as abdominal pain, diarrhea, mucopurulent stool and tenesmus, and is characterized by chronic inflammation of the mucosa of the colon, ulceration of the mucosa or formation of transmural granuloma, which may lead to severe deterioration of the quality of life of the patient, and the patient usually needs to take medicine for life or even surgery for life.

Epidemiological studies have found that IBD is more prevalent in north america and northern europe, but in recent years the prevalence has escalated in the asia-pacific region, with china becoming one of the countries with higher asia prevalence. IBD is better in young and strong years, with an average age of 39 years, and is common in men, with a proportion of 59%. For a particular type of IBD, CD can occur throughout the digestive tract, usually in the terminal ileum and right colon, with lesions that involve the full thickness of the intestinal wall, presenting as segmental, thrush-like ulcers, with necrotic granulomas. UC is frequently found in rectum and sigmoid colon, and the pathological changes involve mucous layer and submucosa, and show continuous and diffuse ulcer with congestion and edema and high recurrence. In addition, the incidence of colorectal cancer of UC patients is obviously increased, which is about 20 times higher than that of common people. Over 20% of patients with UC develop colitis-related colon cancer within 30 years of diagnosis, with mortality rates of more than 50%.

The primary goal in treating IBD patients is to maintain the patient in a state of chronic clinical remission and healing of the sub-mirror intestinal mucosa. The traditional medicine treatment comprises salicylic acid preparation, glucocorticoid, immunosuppressant and the like, wherein the principle is that inflammation is controlled by inhibiting the immune response of the body mostly, although the curative effect of the medicines is immediate, the side effect of the medicines is relatively large, so the medicines are usually not suitable for long-term use, and patients are easy to relapse after stopping taking the medicines. The traditional Chinese medicine is used as a Chinese traditional culture treasure, has a non-negligible curative effect on chronic recurrent inflammation, has small relative side effect and has a wide application value. From the perspective of traditional Chinese medicine, colitis is mostly caused by spleen-kidney deficiency and cold evil invading stomach, and wolfberry has the efficacy of tonifying kidney and replenishing vital essence due to warm nature, so that wolfberry and extracts thereof have the efficacy of treating IBD on the basis of the theory of traditional Chinese medicine. The modern pharmacology separates out Lycium Barbarum Polysaccharide (LBP) which is an active substance of the Lycium barbarum polysaccharide from the Lycium barbarum, and a large number of documents report that the Lycium barbarum polysaccharide has an immunological activity effect and can effectively inhibit inflammatory reaction and promote organism repair. Lycium barbarum glycopeptide (LbGP) is a glycoconjugate with strong immunological activity and anti-aging effect obtained by further separating and purifying LBP, has a molecular weight of 88kD determined by SDS-PAGE, a sugar content of 70%, and has a sugar composition of Ara: GAl: GlC 2.5:1.0:1.0 (molar ratio) and contains other 18 natural amino acids.

Intestinal lesions in the mouse colitis model induced by Dextran Sodium Sulfate (DSS) most closely resemble the human pathological morphology. The establishment of the DSS-induced mouse UC acute phase model has important value for the research on the occurrence and development of UC diseases and the timing and period of drug action. Therefore, the reliability of the research on the lycium barbarum glycopeptide for preventing or treating ulcerative colitis and inhibiting the conversion of the lycium barbarum glycopeptide into chronic enteritis by adopting a DSS mouse colitis acute model is provided.

At present, no report on the aspect of preventing or treating inflammatory bowel diseases by lycium barbarum glycopeptide exists.

Disclosure of Invention

The invention aims to solve the problem of insufficient drugs for preventing or treating inflammatory bowel diseases, which are used clinically at present and have high efficiency, low toxicity and low price, realize the deep development and utilization of natural products by adopting a modern drug research method, and provide the application of the lycium barbarum glycopeptide in preparing the drugs for preventing or treating inflammatory bowel diseases by combining a large amount of pharmacodynamic experimental studies.

In order to achieve the purpose of the invention, the invention adopts the following technical scheme:

use of lycium barbarum glycopeptide in the manufacture of a medicament for the prevention or treatment of inflammatory bowel disease.

Alternatively, in the above use, the inflammatory bowel disease is ulcerative colitis or crohn's disease.

Alternatively, in the above use, the inflammatory bowel disease is an acute phase of inflammatory bowel disease or a chronic phase of inflammatory bowel disease.

Alternatively, in the above uses, the lycium barbarum glycopeptide significantly advances the chronic remission stage of inflammatory bowel disease, maintains remission, significantly alleviates symptoms of acute colitis, and effectively reduces inflammatory response.

Alternatively, in the above use, the medicament comprises a prophylactically or therapeutically effective amount of the lycium barbarum glycopeptide and a pharmaceutically acceptable carrier.

Alternatively, in the above use, the drug is used in combination with other clinically commonly used drugs for preventing or treating inflammatory bowel disease, or the drug is prepared into a pharmaceutical composition with other clinically commonly used drugs for preventing or treating inflammatory bowel disease.

Preferably, the other clinically commonly used drugs for preventing or treating inflammatory bowel disease are selected from one or more of the following: an antibiotic, 5-aminosalicylic acid (5-ASA) (such as sulfasalazine or mesalamine), a corticosteroid (such as prednisone), an immunomodulator (such as azathioprine or methotrexate), or a biologic (such as infliximab, adalimumab, certolizumab or natalizumab).

The medicine disclosed by the invention is combined with other clinically common medicines for preventing or treating inflammatory bowel diseases, or the medicine disclosed by the invention and other clinically common medicines for preventing or treating inflammatory bowel diseases are prepared into a pharmaceutical composition, the medicine disclosed by the invention containing the lycium barbarum glycopeptide has a remarkable synergistic effect on the other clinically common medicines for preventing or treating inflammatory bowel diseases, and the former can remarkably reduce the dosage of the latter and the toxic and side effects of the latter.

In the pharmaceutical composition or in combined use, the dosage ratio of the medicament of the invention to other clinically common medicaments is 10 in parts by weight: 1-1: 10, preferably, the dosage ratio of the medicine of the invention to other clinically common medicines is 10: 1. 9: 1. 8: 1. 7: 1. 6: 1. 5:1. 4: 1. 3: 1. 2: 1. 1: 1. 1: 2. 1: 3. 1: 4. 1: 5. 1: 6. 1: 7. 1: 8. 1: 9 or 1: 10.

when the drug of the present invention is used in combination with other drugs commonly used clinically for the prevention or treatment of inflammatory bowel disease, the former and the latter may be in the same or different dosage forms. Also, the former and the latter may be administered simultaneously or separately.

Alternatively, in the above use, the medicament is an oral preparation, an injection or a rectal administration preparation.

Preferably, the oral preparation is selected from oral liquid, tablets, powder, capsules or granules.

Compared with the prior art, the invention has the following beneficial effects:

the invention combines the advantages of China in the aspect of natural product research, and discovers the new application of the natural product lycium barbarum glycopeptide in the aspect of preventing or treating inflammatory bowel diseases for the first time. The experimental result of the mouse acute ulcerative colitis model induced by Dextran Sodium Sulfate (DSS) shows that the lycium barbarum glycopeptide administration treatment can remarkably advance the chronic remission stage, remarkably relieve the symptoms of acute colitis, effectively reduce inflammatory reaction and show obvious anti-inflammatory effect. The experimental results show that the lycium barbarum glycopeptide serving as the traditional Chinese medicinal material extract product can be effectively used for preventing and treating inflammatory bowel diseases, has a prospect of developing a medicament for preventing and treating inflammatory bowel diseases, provides more treatment options for clinically preventing or treating inflammatory bowel diseases, and has important social benefits and economic values.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:

FIG. 1 is a schematic diagram of the mouse ulcerative colitis model construction and treatment cycle.

FIG. 2 is a line graph showing the body weight change during the mouse ulcerative colitis model construction and lycium barbarum glycopeptide treatment cycle.

FIG. 3 is a bar graph of the difference in disease activity index during the mouse ulcerative colitis model construction and lycium barbarum glycopeptide treatment cycle.

FIG. 4 is a bar graph of colon length during the mouse ulcerative colitis model construction and lycium barbarum glycopeptide treatment cycle.

FIG. 5 is a picture of colon of each group at each time point during the process of constructing mouse ulcerative colitis model and treating lycium barbarum glycopeptide.

FIG. 6 is a photograph of H & E stained pathological sections of various groups of colons (magnification: 200X) after one week of treatment with Lycium barbarum glycopeptide for ulcerative colitis in mice (W2, Day 14).

FIG. 7 shows the relative expression level of colon IL-1 β in each group when mouse ulcerative colitis lycium barbarum glycopeptide is treated for one week (W2, Day 14).

FIG. 8 shows the relative expression level of colon IL-10 in each group of mice treated with the peptide of Lycium barbarum for ulcerative colitis for one week (W2, Day 14).

Detailed Description

The inventor adopts a modern medicine research method to deeply develop and utilize natural products, and discovers that the lycium barbarum glycopeptide can prevent or treat inflammatory bowel diseases, particularly ulcerative colitis and Crohn's disease for the first time through a large amount of screening. The present invention has been completed based on this finding.

The dosage form of the medicament of the invention is preferably an oral dosage form.

The oral dosage form is capsule, tablet, granule or oral liquid. Preferably tablets or capsules.

The pharmaceutically acceptable carrier refers to conventional pharmaceutical carriers in the field of pharmaceutical preparations, and is selected from one or more of fillers, binders, disintegrants, lubricants, suspending agents, wetting agents, pigments, flavoring agents, solvents and surfactants.

Fillers of the present invention include, but are not limited to, starch, microcrystalline cellulose, sucrose, dextrin, lactose, powdered sugar, glucose, and the like; such lubricants include, but are not limited to, magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, poloxamers, and the like; such binders include, but are not limited to, water, ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, sodium alginate, polyvinylpyrrolidone, and the like; such disintegrants include, but are not limited to, starch effervescent mixtures, i.e., sodium bicarbonate and citric acid, tartaric acid, low substituted hydroxypropylcellulose, and the like; suspending agents include, but are not limited to, polysaccharides such as acacia gum, agar, alginic acid, cellulose ethers, carboxymethyl chitin ester, and the like; including but not limited to water, balanced salt solutions, and the like.

The pharmaceutical composition can be prepared into various solid oral preparations, liquid oral preparations and the like. The pharmaceutically acceptable oral solid preparation comprises the following components: common tablet, dispersible tablet, enteric coated tablet, granule, capsule, dripping pill, powder, etc., and oral liquid preparation includes oral liquid, emulsion, etc. The various dosage forms can be prepared according to the conventional process in the field of pharmaceutical preparation.

The lycium barbarum glycopeptide can be extracted and separated from plants such as lycium barbarum containing the active ingredient by adopting a conventional natural product extraction method in the field, and can also be purchased from commercial products.

In the medical applications described herein, the timing, frequency, and the like of administration of the glycopeptide of lycium barbarum will depend on the particular diagnosis of the condition, and is within the skill of one of ordinary skill in the art.

The therapeutic regimen for mice is applied to human body, and the effective dose of all drugs to human body can be converted by the effective dose of the drugs to mice, which is also easy to be realized by the ordinary skilled person in the art.

In order to better understand the essence of the present invention, the following detailed description section uses pharmacodynamic experiments and results to further illustrate the new use of lycium barbarum glycopeptide in the pharmaceutical field for preventing or treating inflammatory bowel disease.

The invention is further illustrated with reference to specific examples. It should be understood that the specific embodiments described herein are illustrative only and are not limiting upon the scope of the invention.

The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products which are not known to manufacturers and are available from normal sources.

The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples are all commercially available products unless otherwise specified.

Example (b): daily gavage administration of lycium barbarum glycopeptide for treating DSS-induced ulcerative colitis

DSS-induced mouse acute colitis models were grouped and established by the method shown in figure 1.

In this pharmacodynamic study, Student's t test and Welch's t test were used to analyze data between the two groups. More than two groups of data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey multiple comparison test. Graphpad Prism 9 (version 0.1.1) and R Studio (version 1.4.1106) were used for statistical analysis. All data are expressed as mean ± SEM. P <0.05 was considered statistically significant. Wherein, P <0.5, P <0.05, P <0.005, P < 0.0001.

C57BL \6 mice (purchased from Beijing Huafukang Biotech GmbH) of 6-8 weeks of age were used and randomly divided into three groups of 15 mice each before the start of molding. The first group was Control group (Control); the second group is DSS-induced colitis model group (DSS); the third group is lycium barbarum glycopeptide treatment group (LbGP). Before the experiment, each group is weighed and numbered. Each group of mice was independently housed, given adequate feed and had free access to water.

DSS-induced modelling was used in the model and treatment groups of mice in the first week (Day0-Day 7). Preparing 3.5% DSS solution, weighing 12g DSS, dissolving in 300mL sterile water, storing at 4 ℃, filling the drinking troughs in the cages of the model group and the treatment group of mice with the DSS solution at least at the dosage of 5mL DSS solution/mouse/Day (at least 200mL per cage) at the beginning of the experiment (Day0), and adding equal volume of drinking water without DSS to the control group of mice. The residual DSS solution in the model group and the treatment group mice drinking troughs is emptied every 2-3 days, and fresh DSS solution is reconfigured and replaced.

After one week of drinking the DSS solution, the molding was completed, and the body weight of the mice was recorded in groups on the seventh Day (Day 7); taking the feces, recording the feces state and observing whether the feces blood exists or not. Each group killed 3 mice, 200 μ l of blood was collected from the eyeball, centrifuged at 3000rpm for 15 minutes, and the upper serum was collected and stored at-80 ℃; the colon from the anus to the ileocecal region was removed, and the length of the colon was measured and photographed. The colon section about 2cm above the anus is cut to about 1cm long, washed with PBS solution and then quickly fixed in 4% paraformaldehyde. The mesenteric lymph nodes of the mice were excised, washed with PBS solution and then quickly fixed in 4% paraformaldehyde. The fixed tissue samples were stored at 4 ℃.

After the molding is finished, the DSS solution in the drinking troughs in the mouse cages of the model group and the treatment group is replaced by normal water. And mice in the treatment group were administered a daily gavage of 10mg/kg lycium glycopeptide solution for a total of three weeks starting from the end of the molding and until the end of the experiment (Day7-Day 28). According to the lycium barbarum glycopeptide dose of 10mg/kg, the lycium barbarum glycopeptide intake of a mouse with the weight of about 20g is 0.2mg per day. Preparing 1mg/mL medlar glycopeptide working solution, weighing 10mg medlar glycopeptide by using a precision balance, dissolving the medlar glycopeptide in 10mL physiological saline, storing at 4 ℃, replacing medlar glycopeptide solution after 2-3 days, and reconfiguring fresh medlar glycopeptide solution and storing at 4 ℃. The mice in the treatment group are subjected to intragastric administration by 0.2mL of medlar glycopeptide working solution through an intragastric administration needle at a fixed time every day, and meanwhile, the mice in the control group and the model group are subjected to intragastric administration by 0.2mL of physiological saline through the intragastric administration needle.

Every seven days, which is the end of a period, samples were taken for testing. Namely, the weights of the mice were recorded in groups at 14 th, 21 th and 28 th days (Day14, Day21 and Day28) after the start of the experiment; taking the feces, recording the feces state and observing whether the feces blood exists or not. Each group sacrificed 3 mice, tail vein blood was collected and stored in an anticoagulation tube, eyeball blood was collected and centrifuged to collect upper serum and stored at-80 ℃; the colon from the anus to the ileocecal part is taken, the length is measured, a picture is taken, a colon section with the length of about 1cm at the position of 2cm at the upper end of the anus is cut, and the colon section is quickly fixed in 4% paraformaldehyde after being cleaned by PBS solution. The mesenteric lymph nodes of the mice were excised, washed with PBS solution and then quickly fixed in 4% paraformaldehyde. The fixed tissue samples were stored at 4 ℃.

Ulcerative colitis causes abdominal pain and thus causes loss of appetite, reduced food intake, combined with diarrhea and other causes, and the main clinical manifestation is weight loss, so the severity of the disease is reflected to a certain extent by the weight loss. The weights (Mean ± SEM, unit g) of the groups recorded at each stage were as follows: before the experiment (Day0), the weight of the control group mouse is 22.06 + -0.28, the weight of the model group mouse is 22.34 + -0.42, and the weight of the administration group mouse is 22.00 + -0.29; after the first week colitis molding is finished (Day7), the weight of a control group mouse is 22.85 +/-0.42, the weight of a model group mouse is 21.58 +/-0.31, and the weight of an administration group mouse is 20.30 +/-0.54; one week after the second week (Day14), the weight of the control group mice is 24.96 + -0.46, the weight of the model group mice is 22.59 + -0.41, and the weight of the administration group mice is 23.90 + -0.35; two weeks after the administration in the third week (Day21), the weight of the control group mice was 25.00 + -0.49, the weight of the model group mice was 20.91 + -0.18, and the weight of the administration group mice was 24.74 + -0.31; three weeks after the administration at the fourth week (Day28), the blank group mice weighed 24.96. + -. 0.43, the model group mice weighed 25.30. + -. 0.45, and the administration group mice weighed 25.37. + -. 0.41. As shown in the body weight change line graph of fig. 2, the body weight of the control mice steadily increased due to normal growth, and leveled off starting at the third week. In the second week, mice in the model group and the administration group gained weight loss (P <0.05) due to DSS-induced intervention. From the second week of administration, the body weight of the mice in the administered group was significantly increased (P <0.001) compared to the previous period due to the intervention of lycium glycopeptide, while the body weight of the model group was not significantly changed (P > 0.05). Starting from the time of administration, the body weight of the administered group began to rise steadily and gradually coincided with that of the control group, indicating that lycium barbarum glycopeptide can significantly reverse the weight loss caused by DSS-induced ulcerative colitis. In the fourth week (Day28), the body weight of the model mice also increased to the control level, possibly due to self-healing of acute colitis, which resulted in decreased abdominal pain and increased feeding, but the self-healing time was much longer than that of drug intervention.

The main clinical symptoms due to colitis include weight loss, persistent diarrhea, bloody purulent stool, loss of appetite, unexplained weight loss, etc. Therefore, the weight loss degree, the stool character and the hematochezia condition of the mice are scored according to the recorded weight of the mice and the observed stool state of the mice to obtain Disease Activity Indexes (DAI) so as to evaluate the disease severity of each group of mice at each stage. The specific scoring criteria are as follows: the weight loss was 0%, and the score was 0; the weight loss is 1-5%, and is marked as 1 point; the weight loss was 5-10%, and was scored as 2 points; the weight loss was 10-15%, and was scored as 3 points; the weight loss was greater than 15%, and was scored as 4 points; the stool has normal properties and is marked as 0 point; loose stool characters are marked as 1 point; the stool is thin stool and is marked as 3 points; feces have no hematochezia and are marked as 0 point; fecal blood was occult blood positive and scored as 1 point; feces and blood were visibly obvious bloody stools and scored as 3 points. Disease activity indices (Mean ± SEM) for each group of mice evaluated at each stage were as follows: after the first week colitis molding is finished (Day7), the DAI index of a control group mouse is 0.13 +/-0.13, the DAI index of a model group mouse is 4.38 +/-0.89, and the DAI index of an administration group mouse is 5.75 +/-0.82; one week after the second week of administration (Day14), the DAI index of the control group mice is 0.13 + -0.13, the DAI index of the model group mice is 2.13 + -0.48, and the DAI index of the administration group mice is 1 + -0.46; two weeks after the third week (Day21), the control group mouse DAI index was 0.25 + -0.16, the model group mouse DAI index was 2.38 + -0.26, and the administration group mouse DAI index was 0.63 + -0.38; three weeks after the administration at the fourth week (Day28), the DAI index of the mice in the blank group was 0.20. + -. 0.20, the DAI index of the mice in the model group was 0.67. + -. 0.49, and the DAI index of the mice in the administered group was 0.00. + -. 0.00. As shown in fig. 3, the data index difference histograms grouped according to different periods show that at the end of the first week of molding, both the model group and the administered group developed severe colitis symptoms (P <0.001), and the disease symptoms decreased overall with time, but after the second week of administration, the data index of the administered group was not significantly different from that of the control group (P0.38), and the data index of the model group was still significantly higher than that of the control group (P <0.01), and the significant difference remained until the last week disappeared. This indicates that lycium barbarum glycopeptide can indeed significantly alleviate colitis symptoms, and that intervention of lycium barbarum glycopeptide can advance the time point when symptoms are transferred well relative to self-healing.

The short colon length is another important manifestation of acute colitis, and ulcerative inflammation in the colon can cause congestion, edema and ulcer of the colon, which leads to the short colon length, as shown in the picture of fig. 5, the whole colon from the cecum to the anus is taken, and the colon is stretched and straightened, and the total length of the colon is obviously different among three groups. The colon lengths (Mean ± SEM, units cm) of each group of mice at each time period were recorded as follows: after the first week colitis molding is finished (Day7), the colon length of the control group mouse is 9.40 +/-0.12, the colon length of the model group mouse is 6.67 +/-0.13, and the colon length of the administration group mouse is 6.07 +/-0.33; one week after the second week of administration (Day14), the colon length of the control group mice is 9.40 + -0.32, the colon length of the model group mice is 7.37 + -0.15, and the colon length of the administration group mice is 9.00 + -0.21; two weeks after the third week (Day21), the colon length of the control group mouse is 9.40 + -0.10, the colon length of the model group mouse is 7.80 + -0.52, and the colon length of the administration group mouse is 9.10 + -0.15; three weeks after administration at the fourth week (Day28), the colon length of the blank group mice was 8.77. + -. 0.19, the colon length of the model group mice was 8.67. + -. 0.15, and the colon length of the administration group mice was 8.90. + -. 0.26. As shown in fig. 4, a bar graph of colon length difference grouped at different time periods, the colon length was significantly decreased (P <0.0001) in the first week (Day7) model group and the administration group after DSS induction treatment compared to the control group. In the first week after administration (Day14), the colon length recovered in the administered group and was not statistically significantly different from that in the control group (P0.50), while the colon shortening was still significant in the model group compared to the control group (P < 0.0005). This statistical difference persists until no significant difference is seen between the last wednesday groups. The lycium barbarum glycopeptide can obviously reverse colon shortening caused by DSS-induced ulcerative colitis. The results are consistent with the weight change and the DAI index, which indicates that the lycium barbarum glycopeptide can obviously shorten the course of disease and obviously advance the recovery period.

The colon tissue and mesenteric lymph node fixed by 4% paraformaldehyde in the above experiment are placed in melted paraffin and insulated, and after the paraffin is completely immersed in the tissue, the tissue is buried, cooled and solidified, and then fixed on a slicer to be cut into 5-8 micron slices. Removing paraffin from the slices by using dimethylbenzene, dehydrating by using absolute ethyl alcohol, washing by using distilled water, staining cell nuclei by using hematoxylin, staining cytoplasm by using eosin, dehydrating and sealing the slices by using ethyl alcohol and dimethylbenzene finally, completing H & E staining, and then placing the slices under a microscope lamp for observation and photographing for knowing the infiltration condition and the damage condition of immune cells of colon tissues. As shown in fig. 6, in the third week, DSS induction caused severe intestinal epithelial injury, and it can be seen from pathological sections that compared with the control group, the DSS model group had a large number of intestinal epithelial surface cells that were exfoliated and clearly ulcerated, had an uneven and abnormal twisted mucosal surface, had an abnormal and decreased number of crypts, had twisted or branched crypts, had partially atrophic crypt cysts, had fewer goblet cells, and had a large number of inflammatory cell infiltrations in the submucosa. Compared with the model group, the administration group after two weeks of administration has relatively complete intestinal epithelial mucosal surface, regular and clear crypt shape, and serious submucosal inflammation infiltration compared with the control group but obviously lighter than the model group, which is enough to prove that the medlar glycopeptide administration obviously reverses the DSS-induced colon pathological injury.

Taking a colon sample frozen and stored at the temperature of minus 80 ℃ in the experiment, homogenizing and fully grinding on ice, adding an RNA extracting solution, centrifuging to take supernatant, extracting mRNA, performing reverse transcription, and measuring the relative expression quantity of the inflammatory factor by a quantitative PCR (polymerase chain reaction) method. The results of the experiment are shown in fig. 7 and 8. The experimental result shows that the lycium barbarum glycopeptide can obviously reduce the expression of proinflammatory cytokine IL-1 beta and obviously increase the expression of an anti-inflammatory cytokine IL-10.

In conclusion, the continuous three-week intragastric administration of lycium barbarum glycopeptide can continuously relieve the symptoms and inflammatory indexes of acute colitis, and compared with the self-healing process of a model group, the administration treatment of lycium barbarum glycopeptide can remarkably advance the chronic remission stage and better recover.

The invention discloses dynamic changes of various indexes of the lycium barbarum glycopeptide in the treatment process of ulcerative colitis, proves a great application of the lycium barbarum glycopeptide as an anti-acute colitis medicament, and shows that the lycium barbarum glycopeptide only needs 10mg/kg to exert the drug effect, improve the colitis symptoms and has no medicament tolerance. In addition, in the animal experiments, no obvious adverse reaction exists in the lycium barbarum glycopeptide, and the experimental animals have good tolerance to the lycium barbarum glycopeptide, which indicates that the lycium barbarum glycopeptide has high safety.

Clinically, a hormonal drug such as aminosalicylic acid preparation or glucocorticoid is generally used for treating inflammatory bowel disease, and the hormonal drug inhibits inflammation by inhibiting in vivo hormone synthesis and immune response. Although the curative effect of the medicines is immediate, the side effects of the medicines are very large, for example, even sulfasalazine with relatively small toxic and side effects in the medicines commonly used in clinic for treating the inflammatory bowel diseases easily causes liver and kidney injury, mental disorder, skin mucosa damage, thrombocytopenia and aplastic anemia and digestive system symptoms of different degrees, such as nausea and vomiting, and the medicines are generally not suitable for long-term use, and the patients easily relapse after stopping taking the medicines.

In recent years, with intensive research on inflammatory bowel diseases, various biological agents originally developed for the treatment of tumors have been used for the treatment of inflammatory bowel diseases, for example, the JAK inhibitors tofacitinib, integrin inhibitors, mabs and interleukin inhibitor eculizumab. However, biological products have high limitations on applicable populations, are very expensive, and are not affordable by the salary family.

Therefore, compared with the chemical drugs and biological agents which are commonly used for treating the inflammatory bowel diseases clinically, the lycium barbarum glycopeptide has unique advantages in the aspects of protecting target organs, controlling inflammatory reaction, improving clinical symptoms, reducing relapse and the like. The lycium barbarum glycopeptide serving as a natural traditional Chinese medicine extract has safe and green sources and small side effects, can regulate the immunity of an organism in many aspects to generate an anti-inflammatory effect, is suitable for continuously taking medicines to inhibit colonic inflammation for a long time and prevent recurrence, has the prospect of developing medicines for preventing and treating inflammatory bowel diseases, provides more treatment options for clinically preventing or treating the inflammatory bowel diseases, and has important social benefits and economic values.

It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims

1. Use of lycium barbarum glycopeptide in the manufacture of a medicament for the prevention or treatment of inflammatory bowel disease.

2. Use according to claim 1, characterized in that: the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

3. Use according to claim 1 or claim 2, characterized in that: the inflammatory bowel disease is an acute phase of inflammatory bowel disease or a chronic phase of inflammatory bowel disease.

4. Use according to any one of claims 1 to 3, characterized in that: the lycium barbarum glycopeptide remarkably advances the chronic remission stage of inflammatory bowel disease, keeps the state of remission of the disease, remarkably relieves the symptoms of acute colitis and effectively reduces inflammatory reaction.

5. Use according to any one of claims 1 to 4, characterized in that: the medicament comprises an effective amount of lycium barbarum glycopeptide for prevention or treatment and a pharmaceutically acceptable carrier.

6. Use according to any one of claims 1 to 5, characterized in that: the medicine is combined with other clinically common medicines for preventing or treating inflammatory bowel diseases, or the medicine and other clinically common medicines for preventing or treating inflammatory bowel diseases are prepared into a pharmaceutical composition.

7. Use according to claim 6, characterized in that: the other clinically commonly used drugs for preventing or treating inflammatory bowel disease are selected from one or more of the following: an antibiotic, 5-aminosalicylic acid (5-ASA) (such as sulfasalazine or mesalamine), a corticosteroid (such as prednisone), an immunomodulator (such as azathioprine or methotrexate), or a biologic (such as infliximab, adalimumab, certolizumab or natalizumab).

8. Use according to any one of claims 1 to 7, characterized in that: the medicine is an oral preparation, an injection or a rectal administration preparation.

9. Use according to claim 8, characterized in that: the oral preparation is selected from oral liquid, tablets, powder, capsules or granules.