CN115414369A

CN115414369A - Application of cucurbitacin C in preparation of medicine for preventing or treating inflammatory bowel disease

Info

Publication number: CN115414369A
Application number: CN202211047704.5A
Authority: CN
Inventors: 杨海霞; 赵彤; 邓建军
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2022-08-30
Filing date: 2022-08-30
Publication date: 2022-12-02
Anticipated expiration: 2042-08-30
Also published as: CN115414369B

Abstract

The invention provides an application of cucurbitacin C in preparing a medicament for preventing or treating inflammatory bowel disease. The cucurbitacin C can effectively prevent or treat the inflammatory bowel disease, is safe to use and good in application prospect, and provides a new idea for preventing and treating the inflammatory bowel disease clinically.

Description

Application of cucurbitacin C in preparation of medicine for preventing or treating inflammatory bowel disease

Technical Field

The present invention relates to the field of medicine. In particular, the invention relates to application of cucurbitacin C in preparation of a medicament for preventing or treating inflammatory bowel disease.

Background

Inflammatory Bowel Disease (IBD) is a chronic Inflammatory disease of the gastrointestinal tract involving the ileum, rectum or colon, including Ulcerative Colitis (UC) and Crohn's Disease (CD). IBD is characterized by a long course of disease, different degrees of severity of disease and a high repetition rate, and patients often have symptoms and signs of persistent abdominal pain, diarrhea, rectal bleeding, weight loss, fatigue, and the like. In recent years, the incidence of IBD has increased year by year, and it is a chronic inflammatory disease that is urgently to be solved.

However, at present, preventive or therapeutic drugs against inflammatory bowel diseases are still under investigation.

Disclosure of Invention

The present invention aims to solve, at least to some extent, the technical problems of the prior art.

The invention provides application of cucurbitacin C in preparation of a medicament. According to an embodiment of the present invention, the medicament is for preventing or treating inflammatory bowel disease. The inventor finds that cucurbitacin C can effectively prevent or treat inflammatory bowel diseases, is safe to use and good in application prospect, and provides a new idea for clinically preventing and treating inflammatory bowel diseases.

According to an embodiment of the present invention, the use of cucurbitacin C in the preparation of a medicament may further have the following additional technical features:

according to an embodiment of the invention, the medicament is for alleviating weight loss due to suffering from inflammatory bowel disease. The inventor finds that the body weight reduction phenomenon of the body suffering from inflammatory bowel disease can be obviously relieved by administering cucurbitacin C.

According to an embodiment of the invention, the medicament is for inhibiting expression of a pro-inflammatory cytokine. By administering cucurbitacin C to an organism, the expression of proinflammatory cytokines can be effectively inhibited.

According to an embodiment of the invention, the pro-inflammatory cytokine is selected from the group consisting of IL-6, IL-1 β, TNF- α, IFN- γ. The inventor finds that cucurbitacin C can effectively inhibit the expression of proinflammatory cytokines.

According to an embodiment of the invention, the medicament is for promoting the expression of an anti-inflammatory cytokine. By administering cucurbitacin C to a body, the expression of the anti-inflammatory cytokine can be effectively promoted.

According to an embodiment of the invention, the anti-inflammatory cytokine is selected from the group consisting of IL-4 and IL-10. The inventor finds that cucurbitacin C can effectively promote the expression of the anti-inflammatory cytokine.

According to an embodiment of the invention, the drug is administered orally, by gavage or by enema.

According to an embodiment of the invention, the drug is administered at a dose of 1-5 μ g cucurbitacin/kg human. Therefore, the composition can effectively prevent or treat inflammatory bowel diseases, and has the advantages of safe use and small side effect.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Drawings

The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:

FIG. 1 shows an analytical graph of the treatment of each experimental group for four weeks, according to one embodiment of the present invention, wherein A: body weight change profile of mice in the first three weeks; b: weight change profile of mice in the fourth week; c: peripheral mouse DAI score plot;

fig. 2 shows an analytical graph of the treatment of each experimental group for four weeks, according to one embodiment of the present invention, wherein a: a picture of a real colon; b: colon length analysis chart;

FIG. 3 shows an analysis of a colon tissue after treatment according to one embodiment of the present invention, wherein A: a section of a mucosal structure of colon tissue; b: colon HS histological score map;

fig. 4 shows an analysis chart of inflammatory factors according to an embodiment of the present invention, in which a: IL-4 level concentration profile; b: IL-6 level concentration profile; c: IL-10 level concentration profile; d: IL-1 beta level profile; e: analysis chart of TNF-alpha level concentration; f: IFN-. Gamma.level concentration profile.

Detailed Description

The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are conventional products which are commercially available, and are not indicated by manufacturers.

Example 1

Male C57BL/6 mice of 5-6 weeks old, weighing 18-22g, 18 mice were randomly divided into 3 groups: blank group (CK group), model group (DSS group), cucurbitacin C (concentration 0.05 mg/kg) intervention group (CuC group). The feeding mode of CK group mice is as follows: free water diet, gavage once daily 0.2mL of 0.5% CMC (carboxymethyl cellulose solution for dissolving CuC); the feeding mode of the mice in the DSS group is as follows: 0.5% CMC of 0.2mL once daily gavage, taken with 2% DSS (sodium dextran sulfate, used to induce colitis) containing water, on the first three weeks free drinking water diet, on the fourth week free drinking water diet; the feeding mode of the mice in the CuC group is as follows: the first three weeks were taken free water diet, gavage of CuC, four weeks were taken free water containing 2% DSS, 0.2mL of cucurbitacin C (0.05 mg/kg concentration solution prepared by dissolution of CuC in CMC) once daily gavage.

The behavior and health of the mice were observed daily, the weight change of the mice was recorded, and Disease Activity Index (DAI) scores were evaluated, and the results are shown in fig. 1. After the experiment, the mice were sacrificed, the colon was removed, and the length of the colon was measured, and the results are shown in fig. 2; preparing a tissue section from the colon tissue of the mouse, and observing the crypt structure and the integrity of the mucous membrane of the colon tissue of the mouse by using H & E staining, wherein the result is shown in figure 3; the plasma of the mice is taken, and the expression levels of proinflammatory cytokines IL-6, IL-1 beta, TNF-alpha and IFN-gamma and the expression levels of the anti-inflammatory cytokines IL-4 and IL-10 in the plasma of the mice are detected by using an ELISA kit, and the result is shown in a figure 4.

As shown in A in figure 1, DSS treatment is not given in the first three weeks, the overall body weight of three groups of mice gradually increases along with the time, and no obvious difference exists among the groups. As shown by B in fig. 1, the rate of change of body weight of mice in DSS group decreased downward from day 4 at the fourth week, and the rate of body weight loss of mice at day 8 was about 12%, which was significantly decreased in body weight (P < 0.0001) compared to mice in the blank group; the mice body weight loss phenomenon caused by colitis can be obviously relieved after cucurbitacin C is dried (P is less than 0.01). The DAI score is an important index for evaluating the severity of IBD, and is calculated according to the comprehensive evaluation of the weight loss percentage of mice, the fecal color, the hardness and the blood-carrying condition of feces. As shown by C in figure 1, on the fourth 4 th day, the feces of the model group mice are loose, brownish red, tired, lying and lazy movement, the body weight is in a descending trend, the feces gradually tend to be unformed and easily adhere to the perianal region, bloody stool visible to the naked eye appears, and the DAI score gradually increases. Through intervention of cucurbitacin C for four weeks, compared with a DSS group, the disease symptoms of mice in the cucurbitacin C intervention group are reduced, and the rising trend of DAI score is remarkably slowed (P is less than 0.01).

The results in FIG. 2 show that: the CK group mice have normal colons and no edema phenomenon, the excrement in the intestinal tract has normal characters, the model group mice have edema of colon intestinal walls, the excrement in the intestinal tract is congested and not formed, the cucurbitacin C intervention group mice have obviously slowed down the edema phenomenon of the colon intestinal walls, the pus and the excrement of blood samples in the intestinal tract are reduced, and the normal excrement of the mice exists; in addition, after the prognosis of cucurbitacin C, the colon length shortening trend of mice in the CuC group is relieved, and the colon length shortening trend is obviously different from that of the DSS group (P < 0.01).

The results in FIG. 3 show that: the mucosa structure of colon tissues of a blank group of mice is not damaged, the shape is normal, the mucosa epithelium is complete, the goblet cells are arranged orderly, the crypt structure exists, no immune cell infiltration is observed in the mucosa layer, after 2 DSS solution is drunk, the colon mucosa structure of a model group of mice is damaged, the crypt structure and the goblet cells disappear, the upper layer structure of the mucosa is incomplete, a large amount of immune cells are infiltrated, after cucurbitacin C intervention, the inflammation of the colon mucosa layer of a colitis mouse is relieved, the goblet cells exist, the colon epithelial cells of the cucurbitacin C intervention group of mice are complete, the tissue injury is obviously improved, and a small amount of immune cell infiltration is locally visible; colon HS scores were significantly lower in mice of the CuC group compared to DSS group (P < 0.01).

The results in FIG. 4 show that: compared with the CK group, the proinflammatory cytokines (IL-6, IL-1 beta, TNF-alpha and IFN-gamma) in the plasma of mice in the DSS group and the CuC group are over-expressed, the expression level of the inflammation-inhibiting cytokines (IL-4 and IL-10) is reduced, but the expression level of the proinflammatory cytokines of the mice in the CuC group is obviously lower than that of the DSS group, and the inflammation-inhibiting cytokines are obviously higher than that of the DSS group. Compared with the CK group, the levels of proinflammatory cytokines IL-6, IL-1 beta, TNF-alpha and IFN-gamma in the plasma of the mice in the DSS group are obviously increased by 31.00%, 25.92%, 32.53% and 14.72% respectively; compared with the DSS group, the plasma concentration of IL-6, IL-1 beta, TNF-alpha and IFN-gamma of mice in the CuC group is respectively reduced by 11.66%, 10.17%, 15.42% and 6.66%, and the two groups have significant difference (P < 0.001). Compared with the CK group, the anti-inflammatory cytokines IL-4 and IL-10 in the plasma of the mice in the DSS group are obviously reduced by 33.87 percent and 31.25 percent respectively; compared with DSS group, the concentration of IL-4 and IL-10 in the plasma of mice in CuC group is reduced by 37.05% and 27.88%, respectively, and the difference between the two groups is significant (P < 0.0001)

In conclusion, the phenomena of weight loss, DAI score increase, colon tissue destruction, over-expression of proinflammatory factors and low expression of inflammation inhibiting factors caused by DSS on mice are effectively relieved by the intervention of the cucurbitacin C, and a theoretical basis is provided for the cucurbitacin C as a medicament for preventing and treating inflammatory bowel diseases.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Moreover, various embodiments or examples and features of various embodiments or examples described in this specification can be combined and combined by one skilled in the art without being mutually inconsistent.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.

Claims

1. Use of cucurbitacin C in the manufacture of a medicament for the prevention or treatment of inflammatory bowel disease.

2. The use according to claim 1, wherein the medicament is for alleviating weight loss due to the development of inflammatory bowel disease.

3. The use according to claim 1, wherein the medicament is for inhibiting expression of pro-inflammatory cytokines.

4. Use according to claim 3, characterized in that the pro-inflammatory cytokines are selected from IL-6, IL-1 β, TNF- α, IFN- γ.

5. The use according to claim 1, wherein the medicament is for promoting the expression of an anti-inflammatory cytokine.

6. The use according to claim 5, wherein said anti-inflammatory cytokine is selected from the group consisting of IL-4 and IL-10.

7. The use according to claim 1, wherein the medicament is administered orally, by gavage or by enema.

8. Use according to claim 1, characterized in that the medicament is administered in a dose of 1 to 5 μ g cucurbitacin/kg human.