Application of amino acid food in constructing constipation animal model and constipation treatment drug and its construction method

Abstract

The present invention relates to the field of biotechnology, and specifically to the use of an amino acid food in constructing a constipation animal model and a drug for treating constipation, as well as methods for constructing the same. The constipation animal model is obtained by feeding an animal an amino acid food, wherein the protein source of the amino acid food is in a hydrolyzed form, and the constipation animal model is constructed by feeding the animal with the amino acid food as the sole food source for at least 1.5 weeks. The amino acid food does not affect the animal's weight and growth and development, has a short modeling time, a stable model, a high success rate, and is consistent in simulating constipation caused by dietary factors in the human population.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to the application of amino acid food in constructing a constipation animal model and a medicine for treating constipation, and a construction method thereof. Background Art

[0002] Constipation is a common gastrointestinal motility disorder, and slow-transmission constipation (STC) is a common type of it. Patients mainly experience difficulty in defecation and / or decreased bowel movement, hard and dry stools, accompanied by abdominal pain, bloating, and decreased appetite. Currently, the prevalence of chronic constipation in Chinese adults is 4% to 10%, and is increasing year by year. The prevalence of chronic constipation in people over 70 years old is as high as 23%. Long-term chronic constipation is closely related to the incidence of colorectal cancer, and it also increases the incidence of ischemic stroke, hypertension, metabolic diseases and other diseases, seriously affecting the quality of life of patients. Clinically, prokinetic agents or laxatives are often used to treat slow-transmission constipation. Severe symptoms may even require surgical resection of the colon, but the treatment effect is generally poor.

[0003] Currently, animal models of chronic constipation are primarily constructed using drugs, including oral administration of compound diphenoxylate and loperamide, which disrupt the intestinal barrier to create a constipation model. However, this acute modeling method cannot better simulate the long-term chronic constipation in humans caused by various factors such as diet and lifestyle. Furthermore, there is no unified standard for drug dosage and duration of administration, resulting in poor model stability and a difficult-to-guarantee success rate, which is inconsistent with the pathogenesis of clinical patients.

[0004] Therefore, establishing a new animal model plays an important role in understanding the pathogenesis, pathophysiological process and drug development of chronic constipation, and has important guiding significance for the clinical treatment of chronic constipation. Summary of the Invention

[0005] In view of existing research and existing problems, the purpose of the present invention is to provide an application of amino acid food in constructing a constipation animal model and a drug for treating constipation, as well as a method for constructing the same.

[0006] The present invention is achieved through the following technical solutions:

[0007] The invention protects the use of amino acid food in constructing a constipation animal model, wherein the protein source of the amino acid food exists in a hydrolyzed form.

[0008] Preferably, the amino acid food further comprises carbohydrates, lipids, dietary fiber, inorganic salts, minerals and vitamins.

[0009] Furthermore, the amino acid food also includes other nutrients.

[0010] Preferably, the amino acid food is hydrolyzed milk powder.

[0011] The present invention also protects a method for constructing the above-mentioned constipation animal model, comprising the following steps:

[0012] S1. The protein source is amino acid food in hydrolyzed form;

[0013] S2: Use the amino acid diet prepared in S1 as the sole food source for the animals for at least 1.5 weeks.

[0014] Preferably, the construction method further comprises step S3: verifying whether the experimental animal is in a constipated state by a colon glass bead excretion test and / or a carmine dye gavage test.

[0015] Furthermore, whether the experimental animals are in a constipated state is determined by whether there is a significant difference compared with animals fed normal food.

[0016] The present invention also protects the use of the above-mentioned constipation animal model in exploring animal intestinal motility and / or animal feces water content.

[0017] The present invention also protects the use of the above-mentioned constipation animal model in screening and / or verifying drugs for preventing, improving and / or treating constipation.

[0018] The present invention also protects the use of the above-mentioned constipation animal model in screening and / or verifying foods or dietary supplements for preventing and improving constipation.

[0019] The beneficial effects of the present invention are:

[0020] The present invention discloses a mouse constipation model induced by an amino acid diet. The amino acid diet does not contain macromolecular antigens, is consistent with the nutritional components of normal food, and does not affect the weight and growth and development of mice. A constipation model can be obtained by feeding the mice with the amino acid diet for two consecutive weeks. The modeling time is short, the model is stable, the success rate is high, it is mild and non-irritating, and it can better simulate the actual situation of patients with chronic constipation in the clinic. It is consistent with constipation caused by dietary factors in the simulated population, providing a new research approach for in-depth exploration of the pathogenesis of chronic constipation and precise treatment of chronic constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The results of the colonic glass bead excretion time of mice in the AFD-induced constipation model (amino acid food group) and the normal food group in Example 2. The numbers marked between the groups in the figure represent the difference between the two groups. P value;

[0022] Figure 2The results of defecation time of mice in the AFD-induced constipation model (amino acid food group) and the normal food group were gavage with carmine dye in Example 2. The numbers marked between the groups in the figure represent the difference between the two groups. P value;

[0023] Figure 3 The results of fecal water content in mice in the AFD-induced constipation model (amino acid food group) and the normal food group in Example 3. The numbers marked between the groups in the figure represent the difference between the two. P value;

[0024] Figure 4 The weight results of mice in the AFD-induced constipation model (amino acid food group) and the normal food group in Example 4. The numbers marked between the groups in the figure represent the difference between the two groups. P value. DETAILED DESCRIPTION

[0025] For a better understanding of the present invention, the present invention is further described below in conjunction with embodiments and drawings. The following embodiments are merely illustrative of the present invention and are not intended to limit the present invention.

[0026] Example 1 Design and synthesis of an amino acid food (Antigen-free diet, AFD)

[0027] This AFD food simulates deep hydrolyzed milk powder and does not contain large molecular antigens. The protein components contained in the food are all hydrolyzed into amino acid form, without changing the original protein ratio and without affecting the form and ratio of other nutrients. The specific formula is shown in Table 1.

[0028] Table 1 Comparison of amino acid diet (AFD) and normal diet (NCD) formula

[0029]

[0030] *. Custom-made and purchased by Nantong Trophy Feed Technology Co., Ltd., the 18 complex amino acids include: glycine, proline, arginine, histidine, lysine, tyrosine, tryptophan, phenylalanine, methionine, cystine, threonine, leucine, isoleucine, valine, glutamic acid, alanine, aspartic acid, and serine;

[0031] **. Purchased from Nantong Trophy Feed Technology Co., Ltd., feed code LAD 0020.

[0032] Example 2 AFD constipation model in mice

[0033] Twelve six-week-old SPF-grade C57 / B6j male mice (purchased from Jicui Yaokang) were randomly divided into two groups: one receiving a normal chow diet (NCD, 6 mice) and the other receiving an amino acid diet (AFD, 6 mice). After two weeks of continuous feeding, their constipation status was monitored using a colonic glass bead test and a carmine dye gavage test. The specific experimental methods are as follows:

[0034] Colon excretion glass bead experiment: Push a 2mm diameter glass bead into the mouse anus, insert a 2mm diameter metal rod to push the glass bead until the glass bead is pushed 2cm into the mouse colon, slowly remove the metal rod, and place the mouse alone in a clean cage. Observe and record the time it takes for the glass bead to be excreted. The longer the excretion time, the weaker the mouse's colon motility.

[0035] Carmine dye gavage experiment: mice were fasted for 12 hours before gavage, and each mouse was gavaged with 200 μL of carmine dye (6% carmine + 0.5% methylcellulose dissolved in sterile water). Then the mice were placed individually in clean cages, and the time when the mice excreted the first red feces was observed and recorded. The longer the excretion time, the weaker the intestinal motility of the mice.

[0036] The experimental results are as follows Figure 1 、 Figure 2 As shown in the figure, the time for the mice in the amino acid food group to excrete glass beads and carmine feces was significantly longer than that in the normal food group, indicating that the intestinal motility of the mice in the amino acid food group was slowed down.

[0037] Example 3 AFD reduces fecal water content in mice

[0038] The fecal moisture content of the mice in Example 2 was measured in the second week. The mice were placed individually in a cage lined with absorbent paper, and the feces were collected and weighed as the wet weight. After freeze-drying, the dry weight was obtained. The fecal moisture content was calculated according to the formula: Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight. The results are shown in the figure. Figure 3 As shown in , the experimental results showed that the fecal water content of mice in the amino acid food group was significantly lower than that in the normal food group.

[0039] Example 4 AFD does not affect the growth and weight gain of mice

[0040] The body weight of mice in the AFD model constructed in Example 2 was tested. Figure 4 As shown, the weight of mice in the amino acid food group was comparable to that of mice in the normal food group, indicating that the amino acid food did not affect the growth and weight gain of mice.

[0041] The above-described embodiments are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the scope of protection determined by the claims of the present invention.

Claims

1. The use of amino acid food in constructing an animal model of constipation is characterized by: The protein source of the amino acid food is in a hydrolyzed form, and the amino acid food is fed continuously for at least 2 weeks as the only food source for the animal; the animal is a mouse; and the amino acid food also includes carbohydrates, lipids, dietary fiber, inorganic salts, minerals and vitamins.

2. The use according to claim 1, characterized in that: The amino acid food is hydrolyzed milk powder.

3. A method for constructing a constipation animal model according to claim 1, characterized in that: The following steps are involved: S1. The protein source is amino acid food in hydrolyzed form; S2: Use the amino acid diet prepared in S1 as the sole food source for the animals for at least 2 weeks.

4. The method for constructing a constipation animal model according to claim 3, characterized in that: The construction method further comprises step S3: verifying whether the experimental animal is in a constipated state by a colon glass bead excretion test and / or a carmine dye gavage test.

5. The method for constructing a constipation animal model according to claim 4, wherein: Whether the experimental animals are in a constipated state is determined by whether there is a significant difference compared with animals fed normal food.

6. Use of the constipation animal model according to any one of claims 1 to 5 in studying animal intestinal motility and / or animal feces water content.

7. Use of the constipation animal model according to any one of claims 1 to 5 in screening and / or verifying drugs for preventing, improving and / or treating constipation.

8. Use of the constipation animal model according to any one of claims 1 to 5 in screening and / or verifying foods or dietary supplements for preventing or improving constipation.

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