CN115414369B

CN115414369B - Application of cucurbitacin C in preparation of medicines for preventing or treating inflammatory bowel disease

Info

Publication number: CN115414369B
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: 2023-09-15
Anticipated expiration: 2042-08-30
Also published as: CN115414369A

Abstract

The invention provides the use of cucurbitacin C in the preparation of a medicament for the prevention or treatment of inflammatory bowel disease. Cucurbitacin C can effectively prevent or treat inflammatory bowel disease, is safe to use and has good application prospect, and provides a new thought for preventing and treating inflammatory bowel disease clinically.

Description

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

Technical Field

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

Background

Inflammatory bowel disease (Inflammatory bowel disease, IBD) is a chronic inflammatory disease of the gastrointestinal tract involving the ileum, rectum, colon, including ulcerative colitis (Ulcerative colitis, UC) and Crohn's Disease (CD). IBD is characterized by a long course, varying degrees of severity and high repetition rate, and patients often have symptoms and signs such as persistent abdominal pain, diarrhea, rectal bleeding, weight loss, and fatigue. In recent years, the incidence of IBD has increased year by year, and chronic inflammatory diseases have become a major issue to be resolved.

However, preventive or therapeutic drugs for inflammatory bowel disease are currently under investigation.

Disclosure of Invention

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

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

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 caused by suffering from inflammatory bowel disease. The inventors found that the body weight loss phenomenon of the body suffering from inflammatory bowel disease can be significantly alleviated by administration of cucurbitacin C.

According to an embodiment of the invention, the medicament is for inhibiting pro-inflammatory cytokine expression. By administering cucurbitacin C to the body, 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. Beta., TNF-alpha, IFN-gamma. The inventors found that cucurbitacin C can effectively inhibit pro-inflammatory cytokine expression.

According to an embodiment of the invention, the medicament is for promoting the expression of an anti-inflammatory cytokine. By administering cucurbitacin C to the body, the expression of the anti-inflammatory cytokines 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 inventors found that cucurbitacin C can effectively promote the expression of anti-inflammatory cytokines.

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

According to an embodiment of the invention, the drug is administered in a dose of 1-5 μg cucurbitacin per kg of human. Therefore, the medicine can effectively prevent or treat inflammatory bowel disease, is safe to use and has 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 foregoing and/or additional aspects and advantages of the invention will become apparent and may be better understood from the following description of embodiments taken in conjunction with the accompanying drawings in which:

fig. 1 shows an analysis chart of four weeks of treatment of each experimental group according to one embodiment of the present invention, wherein a: body weight change plot of mice over the first three weeks; b: fourth week mice weight change profile; c: fourth week mouse DAI score plot;

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

fig. 3 shows an analysis of colon tissue after treatment according to one embodiment of the invention, wherein a: a slice diagram of a colonic tissue mucosa structure; b: colon HS histological score map;

fig. 4 shows an inflammatory factor analysis chart according to an embodiment of the present invention, wherein a: IL-4 level concentration profile; b: IL-6 level concentration profile; c: IL-10 level concentration profile; d: IL-1 beta level concentration profile; e: TNF- α level concentration profile; f: IFN-gamma level concentration profile.

Detailed Description

The scheme of the present invention will be explained below with reference to examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the present invention and should not be construed as limiting the scope of the invention. The examples are not to be construed as limiting the specific techniques or conditions described in the literature in this field or as per the specifications of the product. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

Example 1

The experiment adopts male C57BL/6 mice with the age of 5-6 weeks and the weight of 18-22 g, and 18 mice are randomly divided into 3 groups: blank (CK), model (DSS), cucurbitacin C (concentration 0.05 mg/kg) intervention (CuC). The feeding mode of the CK group mice is as follows: free-drinking diet, 0.2mL of 0.5% cmc (carboxymethyl cellulose solution for dissolving CuC) administered once daily; the feeding mode of mice in the DSS group is as follows: the first three weeks were free to drink, the fourth week was free to drink water containing 2% dss (dextran sodium sulfate for inducing colitis), and 0.2mL of 0.5% cmc was infused once daily; the feeding mode of the CuC mice is as follows: the first three weeks had a free-drinking diet, a stomach-filled CuC, and the fourth week had a free-drinking diet, with water containing 2% dss, and 0.2mL of cucurbitacin C (a solution of 0.05mg/kg prepared by dissolving CuC in CMC) was filled once daily.

Daily observations of mice behaviour and health, recordings of mice weight changes, assessment of disease activity index (Disease activity index, DAI) scores, the results are shown in figure 1. After the experiment is finished, the mice are sacrificed, the colon is taken out, and the colon length is measured, and the result is shown in fig. 2; tissue sections were prepared from the colon tissue of the mice, and the crypt structure and the mucous membrane integrity of the colon tissue of the mice were observed by using H & E staining, and the results are shown in FIG. 3; the expression levels of proinflammatory cytokines IL-6, IL-1 beta, TNF-alpha and IFN-gamma and the expression levels of anti-inflammatory cytokines IL-4 and IL-10 in the plasma of the mice were examined using ELISA kits, and the results are shown in FIG. 4.

As shown in fig. 1 a, the three mice in the first three weeks were not given DSS treatment, and the overall body weight of the mice in the three groups was gradually increased over time, and there was no significant difference between the groups, and the results demonstrated that the experimental animals had no adverse reaction to cucurbitacin C, which had a certain safety. As shown in fig. 1B, DSS group mice showed a decrease in weight change rate from day 4 of the fourth week, and mice on day 8 showed a weight decrease rate of about 12% and a significant decrease in weight (P < 0.0001) compared to the mice in the blank group; the mice weight loss phenomenon (P < 0.01) caused by colonitis can be obviously relieved after the cucurbitacin C is dried. The DAI score is an important index for evaluating the severity of IBD, and is calculated from the comprehensive evaluation of the weight loss percentage, fecal color and hardness, and fecal bloody conditions of mice. As shown in fig. 1C, on day 4 of the fourth week, the model group mice had loose stool, had a reddish brown color, had a tendency to lay tired and lazy, had a tendency to decrease in body weight, had a gradual tendency for the stool to not form, had a tendency to adhere around the anus, and had macroscopic bloody stool, with a gradual rise in DAI scores. After four weeks of cucurbitacin C intervention, the disease symptoms were reduced in mice in the cucurbitacin C intervention group and the trend of increasing DAI scores was significantly slowed (P < 0.01) compared to DSS group.

The results in fig. 2 show that: the colon of the CK group mice is normal, edema phenomenon does not exist, the feces in the intestinal tract are normal, the intestinal tract of the model group mice is edema, the feces in the intestinal tract are congestion and not formed, the phenomenon that cucurbitacin C intervenes on the intestinal tract of the group mice is obviously slowed down, the pus stool in the intestinal tract is reduced, and the normal feces of the mice exist; in addition, after cucurbitacin C dry prognosis, the shortening of colon length tended to be alleviated in CuC mice, with a significant difference (P < 0.01) from DSS mice.

The results in fig. 3 show that: the colon tissue mucous membrane structure of the mice in the blank group is free from damage, normal in morphology, complete in mucous membrane epithelium, ordered in cup cell arrangement, in which the hidden pit structure exists, no immune cell infiltration is observed in mucous membrane layer, after drinking 2% DSS solution, the colon mucous membrane structure of the mice in the model group is destroyed, the hidden pit structure and cup cells disappear, the mucous membrane upper layer structure is incomplete, and a large amount of immune cells infiltrate, after the intervention of cucurbitacin C, the inflammation of the colon mucous membrane layer of the mice in the colonic is relieved, cup cells exist, the colon epithelial cells of the mice in the intervention group of cucurbitacin C are more complete, the tissue injury is obviously improved, and a small amount of immune cell infiltration is locally visible; cuC group mice showed significantly reduced colon HS score (P < 0.01) compared to DSS group.

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

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

In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means 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 present invention. In this specification, schematic representations of the above terms are not necessarily directed 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. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.

While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the 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 suffering from inflammatory bowel disease.

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

4. The use according to claim 3, wherein the pro-inflammatory cytokine is selected from the group consisting of 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 the 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 lavage or by enema.

8. The use according to claim 1, wherein the medicament is administered in a dose of 1-5 μg cucurbitacin/kg human.