A method for constructing a constipation model

Abstract

The application provides a method for constructing a constipation model. The method is as follows: C57BL / 6J mice are selected as experimental animals for pre-feeding, then 2-aminoquinoline is used for intragastrical administration, the intragastrical administration is performed once a day, and the intragastrical administration is performed for a total of 2 weeks, so that an animal constipation model is obtained. The animal constipation model has the substantial advantages of simple operation, low cost, high success rate, strong repeatability and unity with clinical constipation.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of model construction and verification, and particularly relates to a constipation model construction method. BACKGROUND

[0002] Constipation is a common disease, which is manifested as reduced defecation frequency, defecation difficulty, and dry and hard stool. The pathogenesis of constipation is related to abnormal colon motility, abnormal colon smooth muscle, abnormal enteric nervous system (ENS) and enteric neurotransmitter or gastrointestinal hormone, abnormal function of interstitial cells of Cajal (ICC) in the colon, and abnormal expression of aquaporins (AQP). Long-term constipation can cause hemorrhoids, induce cardiovascular and cerebrovascular diseases, increase the incidence of intestinal obstruction, perforation, anal fissure, etc., and also can lead to colorectal cancer. Investigations show that due to the change of dietary structure, the acceleration of life rhythm, the high work pressure, and the influence of other factors such as bad living habits, the prevalence rate of chronic constipation in the general population is 3% to 17%, which increases with age, especially in people over 65 years old, and the incidence rate shows a rising trend year by year, which seriously affects the quality of life of patients. Therefore, the related research on constipation is paid more and more attention and concern by the medical field.

[0003] In modern medicine, animal models are often used for in-depth mechanism research and exploration of the optimal treatment method. The establishment of a constipation animal model highly consistent with the clinic can promote the research on the etiology and pathogenesis of constipation and is of great significance for the screening of new drugs. At present, the main methods for inducing constipation models include drug induction (such as loperamide, compound diphenoxylate, morphine, and rhubarb), physical stimulation (ice water gavage and water restriction), and surgery. However, these external stimuli cannot actually simulate the pathophysiological process of clinical constipation. In addition, the evaluation of the model also lacks effective and comprehensive schemes. At present, the evaluation of the constipation model is mainly through observation of defecation response, gastrointestinal transport function, and colon pathological tissue, which is single and lacks depth. The observation of animal defecation may be subjective, and the single detection of gastrointestinal motility proteins for constipation modeling preparation still needs to be discussed. Therefore, the establishment of a reliable constipation model has great significance for exploring the nature of constipation and developing effective treatment drugs. SUMMARY

[0004] The purpose of the present application is to provide a constipation model construction method.

[0005] The purpose of the present application is also to provide a dose and concentration of a drug for constructing a constipation model.

[0006] To achieve the above technical purposes, the present application adopts the following technical solutions:

[0007] The application discloses a method for constructing a constipation model, and belongs to the field of medical science.

[0008] The experimental animal in the method is a C57BL / 6J mouse with a body weight of 18-20 g, and the experimental mouse is placed in a feeding cage to freely eat and drink water for pre-feeding.

[0009] The concentration of 2-aminoquinoline in the method is 1 mg / ml.

[0010] The dilute solution of 2-aminoquinoline in the method is a PBS solution.

[0011] The application further discloses an animal constipation model constructed by the method.

[0012] Compared with the prior art, the application has the following advantages:

[0013] The application provides a method for constructing a constipation model and application thereof, so as to solve the technical problems that the prior constipation model cannot simulate the pathophysiological process of constipation well, and the evaluation of the constipation model is single, subjective and lacks dynamicity. BRIEF DESCRIPTION OF DRAWINGS

[0014] The accompanying drawings, which form a part of the present application, are intended to provide further understanding of the present application, and the illustrative embodiments of the present application and their description serve the purpose of explaining the present application. In the drawings:

[0015] Figure 1 Fig. 2 is a comparison diagram of stool appearance characteristics of mice in a 2-aminoquinoline group and a normal control group, wherein Control represents the control group, and 2-AQ represents the 2-aminoquinoline group;

[0016] Figure 2 Fig. 3 is a diagram of fecal parameters of mice in the 2-aminoquinoline group and the normal control group, wherein Control represents the control group, and 2-AQ represents the 2-aminoquinoline group;

[0017] Figure 3 Fig. 4 is a comparison diagram of small intestinal transmission results and colon length of mice in the 2-aminoquinoline group and the normal control group, wherein A represents the transmission results of the mice in the 2-aminoquinoline group, B represents the transmission results of the small intestine of the mice in the control group, and C represents the mice in the 2-aminoquinoline group;

[0018] Figure 4 Fig. 5 is a comparison diagram of total horizontal movement distance and total number of movements into the central area of a mine field experiment of constipation mice in the 2-aminoquinoline group and the normal control group;

[0019] Figure 5Figure 2 is a graph of 2-aminoquinoline inhibiting the frequency and amplitude of mouse colon tissue contraction;

[0020] Figure 6 Figure 4 is a bar graph of 2-aminoquinoline constipation mouse model whole gastrointestinal transit time, fecal water content and fecal particles, wherein Control: control group; 2-Aminoquinoline: 2-aminoquinoline; Loperamide: loperamide; Intestinal Transit time determination: whole gastrointestinal transit time; Fecal water content: fecal water content; Fecal particles per hour: fecal particles per hour. DETAILED DESCRIPTION

[0021] The following specific descriptions are exemplary and are intended to provide further explanation of the present application. Those skilled in the art should understand that modifications or substitutions can be made to the details and forms of the technical solutions of the present application without departing from the spirit and scope of the present application, and such modifications and substitutions fall within the protection scope of the present application.

[0022] The experimental animals and chemical reagents used in the present application are commercially available conventional experimental raw materials.

[0023] Example 1: Method for constructing a constipation model

[0024] The experimental animals selected in the present experiment are SPF level 8-week-old male C57BL / 6J mice with a body weight of 18-20 g; 2-aminoquinoline is dissolved in phosphate buffer solution (PBS) and configured to a concentration of 1 mg / ml, and 8-week-old male C57BL / 6J mice are administered by gavage at a dose of 20 mg / kg once a day for 2 weeks. PBS is administered by gavage at the above-mentioned dose as a control group, and modeling and subsequent experiments are performed.

[0025] Example 2: Verification experiment of the constipation model

[0026] After the modeling is completed, the mouse defecation is first evaluated, and the fecal particle number and weight of each group of mice within 30 min are observed and recorded. The fresh feces of the mice are weighed, and then dried to a constant weight at a certain time to calculate the fecal water content of the mice. The fecal size and intestinal motility indicators of the 2-aminoquinoline group and the control group of mice are compared, as shown in Figures 1-2 , and the results show that the fecal water content and fecal particle number of the 2-aminoquinoline group are reduced compared with the control group of mice.

[0027] The mice are administered by gavage with carmine, and the time from gavage to the first red feces is recorded to evaluate the whole gastrointestinal transit time of the mice. The results show that the whole gastrointestinal transit time of the 2-aminoquinoline mice is prolonged compared with the control group of mice. SeeFigure 3 .

[0028] The mice were gently placed in the open field device (50 × 50 × 40 cm) to freely explore for 10 min. The bottom of the box was evenly divided into 16 squares, and the center 4 squares were the center area. The total movement distance and the frequency of entering the center area were considered as indicators of depressive-like behavior. After each test, the field was thoroughly cleaned with 75% ethanol. The animals were placed in the center square to start the measurement, and each measurement lasted for 10 min. The video tracking system (SuperMaze, China) recorded the total horizontal movement distance and the total number of times entering the central area. The results showed that compared with the control group, the total horizontal movement distance and the total number of times entering the central area of the 2-aminoquinoline constipation mice were reduced, as shown in Figure 4 .

[0029] The changes in the movement of the isolated colon were determined. The colon tissues of mice were taken for 2-aminoquinoline perfusion experiments in vitro to evaluate the contraction frequency and amplitude of the colon tissues. The results showed that 2-aminoquinoline could reduce the contraction frequency and amplitude of the isolated colon tissues of normal mice.

[0030] Example 3. Comparative experiments of loperamide constipation model

[0031] According to the model construction method of Example 1, the conventional loperamide was used for intragastrical administration, and the total gastrointestinal transit time, fecal water content and fecal particle number were recorded, and the results are shown in Figure 6 The results showed that from the above detection indexes, it could be seen that the mouse constipation model constructed by intragastrical administration of 2-aminoquinoline could achieve similar effects to the existing loperamide model.

[0032] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method of constructing a model of constipation, characterized by, The construction method comprises the following steps: selecting experimental animals for pre-feeding, and then using 2-aminoquinoline for gavage, once a day, for a total of 2 weeks, so as to obtain an animal constipation model. The concentration of 2-aminoquinoline in the construction method is 1 mg / ml. The experimental animals are mice.

2. The method of claim 1, wherein the constipation model is constructed by, In the construction method, the experimental animals are 8-week-old C57BL / 6J mice with a weight of 18-20 g, and the experimental mice are placed in a feeding cage for free feeding and water drinking for pre-feeding.

3. The method of claim 1, wherein the model of constipation is constructed by: The dilute solution of 2-aminoquinoline in the construction method is a PBS solution. ​

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