Application of garlic polysaccharide in preparation of product for repairing intestinal injury caused by constipation

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of garlic polysaccharide in preparation of a product for repairing intestinal injury caused by constipation. An embodiment verifies that the garlic polysaccharide can effectively repair intestinal physical, chemical and immune barrier injuries caused by constipation, and by up-regulating expression of tight junction proteins (such as ZO-1 and Occludin) and reducing expression of Claudin-1, the garlic polysaccharide can enhance tight junction between intestinal epithelial cells and reduce intestinal permeability; besides, the garlic polysaccharide can promote secretion of intestinal mucoprotein and enhance the protection function of the intestinal mucous layer by up-regulating expression of MUC2 and MUC4 genes. The invention discloses a multi-aspect action mechanism of the garlic polysaccharide in repairing intestinal injury caused by constipation, and the garlic polysaccharide is expected to become a novel intestinal repairing agent for treating constipation and related complications thereof.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and particularly relates to application of garlic polysaccharide in preparing a product for repairing intestinal damage caused by constipation. Background Art

[0002] Constipation is a common digestive symptom, characterized by decreased bowel movement, dry stool, difficulty in defecation, or a feeling of incomplete defecation. It not only affects the patient's quality of life, but may also cause a variety of complications.

[0003] Long-term constipation can lead to slower intestinal motility, feces staying in the intestines for too long, and excessive absorption of water, further aggravating constipation. Feces staying in the intestines for too long can easily breed bacteria, increasing the risk of intestinal infection. Constipation can cause feces to remain in the intestines, increase intestinal pressure, and affect intestinal motility and nerve function. Long-term constipation may also cause intestinal flora imbalance and further damage the intestinal barrier. Studies have shown that constipation can lead to reduced intestinal mucus secretion, reduced expression of mucin (such as MUC2), and down-regulation of tight junction protein expression, thereby increasing intestinal permeability.

[0004] The intestinal barrier is composed of mechanical barriers, chemical barriers, immune barriers and microbial barriers. The mechanical barrier is mainly composed of intestinal epithelial cells and tight junction proteins (such as ZO-1, Claudin-1, Occludin), which prevent the invasion of pathogens and harmful substances. The chemical barrier includes mucus, digestive enzymes and antimicrobial proteins. The immune barrier is composed of intestinal epithelial cells, intraepithelial lymphocytes and intestinal associated lymphoid tissue. The microbial barrier is composed of intestinal flora and its metabolites. The existing technology currently only studies the abnormalities of intestinal flora, and lacks solutions to repair damage to the intestinal mechanical barrier, chemical barrier and immune barrier. Summary of the invention

[0005] The purpose of the present invention is to provide an application of garlic polysaccharide in repairing intestinal damage caused by constipation, and to provide an effective and feasible idea and solution for repairing mechanical barrier, chemical barrier and immune barrier damage caused by constipation.

[0006] The invention provides application of garlic polysaccharide in preparing a product for repairing intestinal damage caused by constipation.

[0007] Preferably, the intestinal damage includes physical barrier damage, chemical barrier damage and immune barrier damage.

[0008] Preferably, the repair comprises increasing the expression of ZO-1 and Occludin and decreasing the expression of Claudin-1.

[0009] Preferably, the repair includes promoting intestinal mucin secretion.

[0010] Preferably, the promoting intestinal mucin secretion includes upregulating the expression of MUC2 and MUC4 genes.

[0011] Preferably, said repair comprises reducing inflammatory cell infiltration.

[0012] Preferably, the repair comprises restoration of colon architecture and restoration of goblet cells.

[0013] Preferably, the repair comprises repairing the muscularis mucosa.

[0014] Preferably, the repair includes upregulating the expression of the anti-inflammatory factor IL-10.

[0015] Preferably, the products include medicines, functional foods and nutritional supplements.

[0016] Preferably, the dosage forms of the drug include capsules, tablets, granules, oral liquids, suspensions, suppositories and sprays.

[0017] Beneficial effects of the present invention:

[0018] The present invention has discovered the application of garlic polysaccharide in repairing intestinal damage caused by constipation. It has been verified by examples that garlic polysaccharide can effectively repair intestinal physical, chemical and immune barrier damage caused by constipation. By up-regulating the expression of tight junction proteins (such as ZO-1 and Occludin) and reducing the expression of Claudin-1, garlic polysaccharide can enhance the tight junctions between intestinal epithelial cells and reduce intestinal permeability; in addition, garlic polysaccharide can also promote the secretion of intestinal mucin and enhance the protective function of the intestinal mucus layer by up-regulating the expression of MUC2 and MUC4 genes.

[0019] Garlic polysaccharide is a natural polysaccharide with good biocompatibility and safety, suitable for long-term use. Its multifaceted role in repairing intestinal damage makes it a promising new intestinal repair agent for the treatment of constipation and its related complications (such as inflammatory bowel disease, intestinal flora imbalance, etc.).

[0020] The present invention reveals the multifaceted mechanism of action of garlic polysaccharide in repairing intestinal damage caused by constipation, which not only provides a new strategy for the treatment of constipation and related diseases, but also lays a foundation for in-depth research on the relationship between intestinal flora and overall health. Its natural, safe and multi-effect characteristics make it have broad application prospects and clinical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0022] Figure 1 Flowchart illustration of the experimental design for the present invention.

[0023] Figure 2 is the relative expression level of intestinal tight junction protein mRNA; A represents the relative expression level of ZO-1 mRNA, B represents the relative expression level of Claudin-1 mRNA, and C represents the relative expression level of Occludin mRNA;

[0024] Figure 3 The first black stool time and the total number of stools of each group of mice; A represents the first black stool time of each group of mice, and B represents the total number of stools of each group of mice;

[0025] Figure 4 The effect of garlic polysaccharide on the contents of mucin MUC2 and MUC4 in the colon of mice; A is the content of MUC2, and B is the content of MUC4;

[0026] Figure 5 The following are pathological sections of the colon of mice in the MOD group and the GPM group under a 200x microscope;

[0027] Figure 6 is the level of anti-inflammatory factor IL-10 in the serum of mice in each group.

[0028] exist Figure 2 to Figure 4 and Figure 6 In the table, lowercase letters indicate significant differences. DETAILED DESCRIPTION

[0029] To further illustrate the present invention, the application of garlic polysaccharide provided by the present invention in preparing a product for repairing intestinal damage caused by constipation is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0030] Example 1

[0031] according to Figure 1 Design the experiment according to the process shown.

[0032] BALB / c male mice were raised for 1 week to adapt to the experimental environment, and then randomly divided into blank control group (CON), constipation model group (MOD), lactulose positive control group (PC), garlic polysaccharide low-dose group (GPL), garlic polysaccharide medium-dose group (GPM), and garlic polysaccharide high-dose group (GPH), with 20 mice in each group. The mice in the MOD group, PC group, GPL group, GPM group, and GPH group were constructed with loperamide method for constipation model. After successful modeling, they were treated as follows:

[0033] GPL group: Garlic polysaccharide was administered to mice by intragastric administration daily at a dose of 1.25 g / kg·bw;

[0034] GPM group: Garlic polysaccharide was administered to mice by intragastric administration daily at a dose of 2.5 g / kg·bw;

[0035] GPH group: Garlic polysaccharide was administered to mice by intragastric administration every day, with the intragastric dose of 5 g / kg·bw;

[0036] PC group: mice were intragastrically gavaged with lactulose daily at a dose of 2.5 g / kg·bw;

[0037] The mice in the CON and MOD groups were gavaged with the same dose of sterile water every day.

[0038] The weight changes of mice were recorded every day. After three weeks of intervention, the mice were fasted for 12 hours and then killed. The serum, colon, feces, etc. of the mice were taken for measurement. The intestinal damage and repair were analyzed. During the gavage period, the mice were free to drink water and eat. The purity of the garlic polysaccharide used for gavage was >90%.

[0039] Serum, colon, colon contents, and fecal samples were sent for testing. Three groups of samples were prepared from each part of each group of mice (12 samples), as well as fecal samples retained at the beginning of the experiment (3 samples), to reveal the intestinal barrier damage and repair of mice under different treatments or environments.

[0040] Physical barrier damage and repair conditions Figure 2 shown.

[0041] Figure 2 A, B, and C are the relative mRNA expressions of intestinal tight junction proteins ZO-1, Claudin-1, and Occludin, respectively. It can be seen that compared with the CON group, the gene expression of tight junction proteins ZO-1 and Occludin in the MOD group was significantly reduced (P<0.05), and the gene expression of Claudin-1 was significantly increased (P<0.05). Mice given three doses of GP can regulate the gene levels of these proteins, increase the gene levels of ZO-1 and Occludin, and reduce the gene level of Claudin-1. The GPH group performed best. There was no significant difference in the gene expression of tight junction proteins ZO-1 and Claudin-1 in the GPH group compared with the PC group, and the gene expression of tight junction proteins in both groups was close to that of the CON group.

[0042] Chemical barrier damage and repair Figure 3-4 shown.

[0043] like Figure 3As shown in the figure, compared with the CON group, the time of the first black stool in the MOD group was significantly increased (P<0.05), indicating that the constipation model was successfully established. After intragastric administration of GP, the time of the first black stool was significantly reduced (P<0.05), especially in the GPH group. The total number of stool pellets in the MOD group was significantly reduced in 5 hours compared with the CON group (P<0.05), indicating that the constipation model was successfully established in mice. Compared with the MOD group, the total number of stool pellets in the GPM and GPH groups was significantly increased (P<0.05).

[0044] Figure 4 Compared with the CON group, the expression of MUC2 and MUC4 genes in the MOD group was significantly downregulated (P < 0.05). This indicates that constipation can cause a decrease in the secretion of intestinal mucin. The GP group significantly upregulated the expression of MUC2 and MUC4 genes (P < 0.05), and the GPH group had the best effect.

[0045] Immune barrier damage and repair Figure 5-6 shown.

[0046] Figure 5 The pathological section of the colon under a 200x microscope is shown. Figure 5 It can be seen that the colon sections of mice in the MOD group showed a large number of inflammatory cells (black arrows) infiltration, some crypts (green arrows) disappeared, goblet cells (red arrows) decreased, and the muscularis mucosa was damaged and unevenly distributed. In the GP group, the colon structure and goblet cells were restored, with a small number of infiltrating inflammatory cells, and the thickness of the muscularis mucosa remained intact.

[0047] Depend on Figure 6 It can be seen that compared with the mice in the CON group, the level of serum anti-inflammatory factor IL-10 in the constipated mice in the MOD group was significantly reduced (P < 0.05). After GP intervention, the level of serum IL-10 increased significantly (P < 0.05), and the GPH group performed best.

[0048] The Dietary Guidelines for Chinese Residents recommends that humans consume 15g of dietary fiber per day. The oral dose of GP for mice is calculated by converting the animal's body weight to the equivalent dose of human body weight.

[0049] It can be seen from the above embodiments that garlic polysaccharide increases the relative expression of intestinal tight junction proteins ZO-1 and Occludin mRNA, reduces the relative expression of Claudin-1 mRNA, and repairs physical barrier damage; after intervention with garlic polysaccharide, the time for the first black stool in mice is shortened, the symptoms of constipation are alleviated, and it can effectively regulate gastrointestinal peptides and neurotransmitters, increase the expression of MUC2 and MUC4 genes, and repair chemical barrier damage; garlic polysaccharide increases the level of anti-inflammatory factor IL-10, repairs immune barrier damage, helps alleviate intestinal barrier damage caused by constipation, alleviates constipation symptoms to a certain extent, and prevents constipation from worsening.

[0050] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of garlic polysaccharide in the preparation of products for repairing intestinal damage caused by constipation.

2. The use according to claim 1, characterized in that: The intestinal damage includes physical barrier damage, chemical barrier damage and immune barrier damage.

3. The use according to claim 1, characterized in that: The repair includes increasing the expression of ZO-1 and Occludin and decreasing the expression of Claudin-1.

4. The use according to claim 1, characterized in that: The repair includes promoting intestinal mucin secretion.

5. The use according to claim 4, characterized in that: The promotion of intestinal mucin secretion includes upregulating the expression of MUC2 and MUC4 genes.

6. The use according to claim 1, characterized in that: The repair includes reducing inflammatory cell infiltration.

7. The use according to claim 1, characterized in that: The repair includes restoration of colon architecture and restoration of goblet cells.

8. The use according to claim 1, characterized in that: The repair includes repairing the muscularis mucosa.

9. The use according to claim 1, characterized in that: The repair includes upregulating the expression of the anti-inflammatory factor IL-10.

10. The use according to claim 1, characterized in that: The products include medicines, functional foods and nutritional supplements; the dosage forms of the medicines include capsules, tablets, granules, oral liquids, suspensions, suppositories and sprays.

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