Spleen and stomach qi deficiency type gastritis animal model and construction method and application thereof

Abstract

The invention discloses a spleen and stomach qi deficiency type gastritis animal model as well as a construction method and application thereof. The construction method comprises the following steps: performing exhaustive swimming exercise on an experimental animal and performing induction by orally taking ammonia water instead of daily drinking water. The clinical symptoms and signs of the animal model constructed by the construction method are similar to those of the spleen and stomach qi deficiency type gastritis, the animal model can be applied to research on the spleen and stomach qi deficiency type gastritis in traditional Chinese medicine, and an experimental basis can be provided for developing related new drug research and development.

Description

Technical Field

[0001] The present invention relates to the technical field of animal model construction, and in particular to an animal model of spleen and stomach qi deficiency type gastritis and a construction method and application thereof. Background Art

[0002] Chronic gastritis (CG) is the most common clinical disease of the digestive system. It refers to gastric mucosal damage caused by an imbalance of invasive and defensive factors. Its essence is that the gastric mucosal structure changes after repeated damage to the gastric mucosal epithelium, which eventually leads to irreversible atrophy or even the disappearance of the intrinsic gastric glands. According to statistics, the prevalence of CG in patients undergoing gastroscopy in my country is about 90%. The persistence and recurrence of chronic non-atrophic gastritis (CNAG) may lead to the development of chronic atrophic gastritis (CAG), which is characterized by gastric mucosal atrophy, often accompanied by metaplasia or dysplasia. As the first step in the malignant transformation of chronic gastritis, CAG is considered to be a precancerous lesion of gastric cancer. There is an urgent need to treat chronic gastritis to prevent its transformation to gastric cancer. At present, the treatment of CG often adopts monotherapy or combination therapy. There are many kinds of Western medicines for the treatment of chronic gastritis, such as anti-gastric acid synthesis, elimination of Hp, anti-intestinal metaplasia and dysplasia, and protection of gastric mucosa. Due to the complex pathogenesis of CG, most Western medicines only partially relieve symptoms, and the symptoms of CG are prone to relapse after discontinuation of medication. In addition, Western medicines also have serious adverse reactions. A considerable number of patients in China have focused their attention on traditional Chinese medicine treatment.

[0003] As one of the most important and oldest complementary medicines in the world, traditional Chinese medicine has formed a unique etiology and diagnostic theory since ancient times. Traditional Chinese medicine divides chronic gastritis into categories such as fullness and stomach pain, and can be divided into five categories: insufficient stomach yin, liver and stomach disharmony, spleen and stomach dampness and heat, spleen and stomach deficiency and cold, and spleen and stomach qi deficiency. Among them, spleen and stomach qi deficiency accounts for a higher proportion.

[0004] The classification of diseases and syndromes in traditional Chinese medicine often comes from the four examinations, which mainly include the main symptoms, secondary symptoms, tongue image, and pulse image. And these indicators are mostly obtained through "looking", "smelling", "listening" and "cutting". The main symptoms of gastritis and spleen and stomach qi deficiency are fullness in the stomach or dull stomach pain, which worsens after meals, and fatigue; secondary symptoms include poor appetite, cold limbs, and loose stools; the pulse is pale tongue or with tooth marks, thin white tongue coating; the pulse is weak. The cause of the disease is mostly due to excessive fatigue and excessive consumption of spicy and cold foods. However, it is difficult to achieve scientific diagnosis through the four examinations in the animal model of traditional Chinese medicine diseases and syndromes, which is not conducive to the screening of drugs for the treatment of traditional Chinese medicine diseases and syndromes. In order to more objectively evaluate the animal model of traditional Chinese medicine diseases and syndromes, the newly constructed animal model of diseases and syndromes is combined with clinical symptoms to find out the objective and measurable indicators corresponding to the diseases and syndromes, and this is used as a preliminary criterion for judging whether the animal model of traditional Chinese medicine diseases and syndromes is successful. The disease-syndrome animal model combines the TCM syndrome model with the Western medicine pathology model. It replicates the disease and syndrome model based on the TCM etiology and Western medicine etiology. It not only reflects the close connection between disease and syndrome, but also conforms to clinical reality and can better reflect the development and changes of the disease. Therefore, the establishment of an animal model of spleen and stomach qi deficiency gastritis is of great significance for further exploring the TCM syndrome of chronic gastritis.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide an animal model of spleen and stomach qi deficiency type gastritis and a construction method and application thereof. The animal model of spleen and stomach qi deficiency type gastritis obtained by the construction method of the present invention can be applied in the disease research of chronic gastritis.

[0007] The present invention is achieved in that:

[0008] In a first aspect, the present invention provides a method for constructing an animal model of spleen and stomach qi deficiency type gastritis, which comprises inducing the experimental animals to undergo exhaustive swimming exercise and to orally take ammonia water instead of daily drinking water.

[0009] In some embodiments, the above-mentioned experimental animals include rats.

[0010] In some embodiments, the exhausting swimming exercise is performed 1-3 times a day.

[0011] In some embodiments, the concentration of the ammonia water is 0.1-0.3 wt %.

[0012] In some embodiments, the induction period is 2-3 months.

[0013] In some embodiments, the above construction method also includes testing the body weight, food intake and water intake of the experimental animals during the induction period, and performing rectal temperature and palm temperature testing, endurance testing, respiratory amplitude and pulse amplitude testing, coagulation analysis, and gastric pathology testing on the experimental animals after the induction.

[0014] In a second aspect, the present invention also provides an animal model of spleen and stomach qi deficiency type gastritis obtained by the above construction method.

[0015] In a third aspect, the present invention also provides the use of the above-mentioned spleen-stomach-qi deficiency type gastritis animal model as an animal model in the research of spleen-stomach-qi deficiency type gastritis.

[0016] In a fourth aspect, the present invention also provides the use of the above-mentioned spleen-stomach-qi deficiency type gastritis animal model in screening drugs for spleen-stomach-qi deficiency type gastritis.

[0017] In a fifth aspect, the present invention also provides the use of the above-mentioned spleen-stomach-qi deficiency type gastritis animal model in evaluating the effect of drugs on spleen-stomach-qi deficiency type gastritis.

[0018] The present invention has the following beneficial effects:

[0019] The present invention constructs an animal model of spleen and stomach qi deficiency type gastritis by exhaustive swimming exercise of experimental animals and oral ammonia water instead of daily drinking water. The animal model constructed by the construction method of the present invention is similar to the clinical symptoms and signs of spleen and stomach qi deficiency type gastritis. The animal model can be applied to the study of spleen and stomach qi deficiency type gastritis in traditional Chinese medicine, and can provide an experimental basis for the development of related new drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0021] Figure 1 The body weight change curves of rats in the normal control group, ammonia water group and model group in Example 1;

[0022] Figure 2 Comparison of food intake and water intake of rats in the normal control group, ammonia water group and model group in Example 1;

[0023] Figure 3 Comparison of the paw temperature, anal temperature and hind paw blood color of rats in the normal control group, ammonia water group and model group in Example 1;

[0024] Figure 4 Comparison of the exhaustive swimming time of rats in the normal control group, ammonia water group and model group in Example 1;

[0025] Figure 5 Comparison of the coagulation index test results of rats in the normal control group, ammonia water group and model group in Example 1;

[0026] Figure 6The stomach pathological pictures of rats in the normal control group, ammonia water group and model group in Example 1;

[0027] Figure 7 The tongue tissue pathological pictures of rats in the normal control group, ammonia water group and model group in Example 1;

[0028] Figure 8 The results of the disease index detection of rats in the normal control group, model group and drug-treated group in Example 1, wherein A: swimming time of rats; B: pulse intensity of rats; C: temperature of the hind paw of rats;

[0029] Fig. 9 The stomach pathological pictures of rats in the normal control group, model group and drug-treated group in Example 1;

[0030] Fig.10 These are the pathological pictures of rat tongue tissues in the normal control group, model group and drug-treated group in Example 1. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0032] Clinically, more than 80% of gastritis is related to Helicobacter pylori infection, which is the most important cause of chronic gastritis. When Helicobacter pylori infection occurs in the stomach, the urease produced decomposes urea to produce ammonia, which can damage the mucosal barrier through multiple pathways to cause gastritis. In traditional Chinese medicine, gastritis is related to exogenous evil, so the inventors of the present invention use oral ammonia water as one of the factors inducing the model; and because excessive fatigue is the main cause of inducing spleen and stomach qi deficiency, the inventors of the present invention establish a spleen and stomach qi deficiency type gastritis model by combining exhausting swimming and oral ammonia water.

[0033] The classification of diseases and syndromes in traditional Chinese medicine often comes from the four examinations, which mainly include the main symptoms, secondary symptoms, tongue image, and pulse image. These indicators are mostly obtained through "looking", "smelling", "listening" and "cutting". The diagnosis results may vary depending on the level of medical skills. It is even more difficult to achieve scientific diagnosis in animals. Therefore, in order to more objectively evaluate the animal model of traditional Chinese medicine diseases and syndromes, combined with the clinical symptoms of spleen and stomach qi deficiency gastritis, the following table is summarized to find out some objective and measurable indicators corresponding to the disease and syndrome, and use this as a preliminary standard for judging whether the animal model of traditional Chinese medicine diseases and syndromes is successful, as shown in the table:

[0034] Table 1 Judgment criteria

[0035]

[0036] When constructing an animal model of spleen and stomach qi deficiency type gastritis, the present invention not only adopts a method combining exhaustive swimming and oral ammonia water to establish the spleen and stomach qi deficiency type gastritis model, but also conducts traditional Chinese medicine counter-evidence on the animal model after construction, wherein the counter-evidence adopts Xiangsha Liujunzi Decoction, which is a decoction composed of Codonopsis pilosula, Atractylodes macrocephala, Poria cocos, Pinellia ternata, Tangerine peel, Magnolia officinalis, Amomum villosum, and Radix Glycyrrhizae Radix, and is mainly used to treat spleen and stomach qi deficiency.

[0037] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0038] Experimental animals and materials used in the examples

[0039] (1) Animals

[0040] 72 SPF SD rats, male, weighing 180-220 g, were provided by Jiangxi University of Traditional Chinese Medicine.

[0041] (2) Materials

[0042] 28% ammonia water (Sigma); isoflurane; hematoxylin; eosin; sodium citrate; normal saline; coagulation analysis reagent kit (Mindray).

[0043] (3) Experimental instruments

[0044] Pulse oximeter, anesthesia machine; paraffin embedding machine (Leica); RM2126 paraffin slicer (Leica); coagulation analyzer (Mindray), balance.

[0045] Example 1

[0046] This embodiment provides a method for constructing an animal model of spleen and stomach qi deficiency type gastritis, which is as follows:

[0047] 1. Modeling method

[0048] (1) Rats were housed in the usual manner for 3 days, with 2-5 rats per cage, room temperature of 20-25°C, relative humidity of 40%-70%, artificial light, 12-h dark cycle, ventilation ≥15 times per hour, and free access to food and water.

[0049] (2) Model construction Rats were randomly divided into three groups: normal control group, ammonia water group and model group, with 12 rats in each group. Rats in each group were allowed to eat freely.

[0050] The normal control group ate and drank normally every day; the model group swam to exhaustion (the rat sank to the bottom of the water for 10 seconds and could not swim out of the water on its own) twice a day, and 0.1% ammonia water replaced drinking water for a total of two months.

[0051] (3) Chinese medicine counter-evidence Rats were randomly divided into 3 groups: normal control group, model group and positive drug group, with 12 rats in each group. The positive drug group was induced with the model in step (2) in the first two months as in the model group. After the model was successfully established, Xiangsha Liujunzi Decoction was gavaged for one week.

[0052] 2. Index detection method

[0053] (1) Body weight: The body weight changes of rats were recorded every week.

[0054] (2) Food intake and water intake: The rats were given a certain weight of feed and a certain volume of drinking water one day before the measurement. After 24 hours, the remaining weight of feed and volume of drinking water were recorded. The difference between the two was the food intake and water intake of the rats in 24 hours.

[0055] (3) Detection of rat respiratory amplitude and pulse amplitude: On the day of the test, no exhaustive swimming model was performed. An anesthesia machine pulse oximeter was used to detect the pulse intensity, respiratory intensity, respiratory rate, heart rate, and blood oxygen concentration of the rats under anesthesia.

[0056] (3) Rectal and hind paw temperatures: On the day of testing, no exhaustive swimming model was performed. A forehead thermometer was used to measure the palm of the right hind limb of the rat under anesthesia for three times, and the average value was taken. Rectal temperature was measured using a temperature probe inserted into the anus of the rat under anesthesia.

[0057] (4) Coagulation analysis: Blood was collected from the ophthalmic vein of rats and anticoagulated with 4% sodium citrate (whole blood: anticoagulant = 9:1). The blood was centrifuged at 3000 r / min for 10 min, and the supernatant plasma was collected. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (FIB) were measured using a coagulation analyzer.

[0058] (5) Rat endurance test: The water temperature was 23±2℃ and the water depth was not less than 60 cm. The rats were placed in the water and swam until they were exhausted. The swimming time was recorded if the rats sank to the bottom of the water for 10 seconds and could not swim out of the water on their own.

[0059] (6) Gastric pathological observation: All rats were killed by cervical dislocation and their stomachs were removed by laparotomy. The stomachs were cut open along the greater curvature of the stomach and flushed with saline. The stomachs were then fixed in paraformaldehyde for pathological observation of gastritis.

[0060] 3. Statistical analysis

[0061] Graphpad prsim5.0 software was used for statistical analysis and plotting. Data were expressed as mean ± standard deviation (Mean ± SD) after normality test. T test and One-way ANOVA test were used for comparison among groups. P < 0.05 was considered statistically significant.

[0062] The experimental results of the animal modeling stage are as follows:

[0063] (1) Body weight, water intake and food intake

[0064] During the modeling process, the weight changes of rats were as follows: Figure 1 As shown in the figure: During the modeling process, ammonia water instead of drinking water did not lead to a decrease in the weight of rats. The weight gain of the spleen and stomach qi deficiency group was significantly lower than that of the control group and the ammonia water group (P < 0.05). After 8 weeks of modeling, the food intake of rats in the model group decreased (P < 0.05), and the water intake remained unchanged. Figure 2 shown.

[0065] (2) Spleen and stomach qi deficiency can lead to a significant decrease in the anal temperature and palm temperature of rats

[0066] After 8 weeks of model induction, there was no statistical difference in palm temperature and rectal temperature between the ammonia group and the control group; however, the palm temperature of the right hind limb of the model group decreased significantly (P<0.01); the rectal temperature also decreased slightly, and there was a statistical difference (P<0.05). Figure 3 As shown in Figure A; it was also observed that the hind paws of the rats in the model group were significantly paler and bloodless compared with those in the control group. Figure 3 As shown in Figure B.

[0067] (3) Swimming time to exhaustion

[0068] After modeling, in order to verify whether the rats in the spleen and stomach qi deficiency group had symptoms of limb fatigue, the endurance of the rats was tested by exhaustive swimming. Figure 4 The results showed that there was no significant difference in endurance between the control group and the ammonia group, while the exhaustive swimming time of the rats in the model group was significantly reduced compared with the two groups (P<0.01).

[0069] (4) Pulse strength

[0070] Compared with the control group, the various indicators of the ammonia group rats were not significantly different from those of the control group (P>0.05), while the heart rate, pulse strength and respiratory intensity of the Qi deficiency group were lower than those of the control group (P<0.05 or P<0.01). The results are shown in Table 2:

[0071] Table 2 Pulse oximeter test indicators of rats in each group

[0072] Blood oxygen concentration Heart rate Respiratory rate Pulse strength Breathing intensity Control group 94.27±3.51 360.69±24.82 76.90±23.28 50.65±18.79 14.30±3.94 Ammonia group 95.19±1.90 355.50±18.90 73.19±16.63 57.83±19.48 18.20±7.96 Model Group 96.93±1.14* 315.38±21.41** 68.75±48.49 25.94±6.62** 4.71±2.35**

[0073] (5) Coagulation index

[0074] like Figure 5 Compared with the control group, the PT and TT of the coagulation indexes of the rats in the model group were significantly shortened (P<0.001 and P<0.05), while there was no significant change in the ammonia group compared with the control group.

[0075] (6) Gastric tissue

[0076] like Figure 6 As shown in the figure, the overall mucosal epithelium of the control group was relatively intact, with no obvious epithelial cell shedding, abundant glands in the lamina propria, visible chief cells and parietal cells, no obvious glandular damage, and no obvious inflammatory cell infiltration; slight mucosal epithelial shedding (black arrow) was observed in the ammonia group, the lamina propria glands were abundant, chief cells and parietal cells were visible, no obvious glandular damage was observed, no obvious inflammatory infiltration was observed in the lamina propria, and there were scattered lymphocyte infiltrations in the local submucosal layer; the overall mucosal epithelium of the model group was relatively intact; some local glandular epithelial cells were necrotic (black arrow), and the nucleus was condensed; scattered eosinophil infiltration was observed at the bottom of the lamina propria. It can be seen that oral administration of ammonia for 2 months induced the occurrence of gastritis, and inflammatory cell infiltration appeared in the stomach of rats in the ammonia group and the model group, but the degree was relatively mild.

[0077] (7) Tongue appearance and tongue tissue

[0078] The tongue of spleen and stomach qi deficiency gastritis is characterized by thin white fur. Figure 7 As shown in the figure, the tongue coating of rats in the control group and ammonia group was pink, while the tongue coating of rats in the model group was thin and white. Further, HE staining of tongue tissues showed that the epithelial structure of the control group and ammonia group was intact, the epithelial cells were morphologically normal and tightly arranged, and no obvious inflammation was observed in the lamina propria; the epithelial structure of the tongue tissue of rats in the model group was intact, the epithelial cells were morphologically normal and tightly arranged, and connective tissue hyperplasia was observed in the lamina propria (black arrow in the lower right corner), accompanied by a small amount of neutrophil infiltration (black arrow in the middle right); the glands were locally atrophied and replaced by hyperplastic connective tissue (black arrow in the upper left corner), a small amount of cells were necrotic, nuclear fragmentation or dissolution (black arrow in the middle left); and round vacuoles of varying sizes were observed in the cytoplasm of many parenchymal cells.

[0079] The experimental results of the TCM disproof stage are as follows:

[0080] (1) Effect of oral administration of Xiangsha Liujunzi Decoction on improving symptoms

[0081] After the model was successfully established, the rats in the drug group began to be gavaged with Liujunzi Decoction once a day. During the drug administration period, the rats in the model group and the drug group changed from swimming twice a day to swimming once every two days. After one week of drug administration, the swimming time, pulse strength and palm temperature of the drug group were significantly improved compared with those of the model group (P<0.001 and P<0.05). Figure 8 shown.

[0082] (2) The improving effect of Xiangsha Liujunzi Decoction on gastric tissue

[0083] like Fig. 9As shown in the figure, the overall mucosal epithelium of the control group was relatively intact, with no obvious epithelial cell shedding, abundant glands in the lamina propria, visible chief cells and parietal cells, no obvious glandular damage, and no obvious inflammatory cell infiltration; the overall mucosal epithelium of the model group was relatively intact, with local submucosal edema (black arrows), scattered inflammatory cell infiltration, and scattered eosinophil infiltration in the local submucosal layer and the bottom of the lamina propria; the local mucosal epithelium of the positive drug group was shedding (black arrows); the number of glands in the lamina propria was abundant, with visible chief cells and parietal cells, no obvious glandular damage, no obvious inflammatory infiltration in the mucosal lamina propria, and scattered eosinophil infiltration in the local submucosal layer. It can be seen that Xiangsha Liujunzi Decoction can alleviate the inflammation of gastritis of spleen and stomach qi deficiency type.

[0084] (3) The improving effect of Xiangsha Liujunzi Decoction on tongue appearance and tongue tissue

[0085] Xiangsha Liujunzi decoction improved the tongue image and tongue tissue inflammation in rats with spleen and stomach qi deficiency type gastritis. Fig.10 The tongue coating changed from thin white to normal. HE staining of tongue tissue showed that the epithelial structure of the model group was intact, the epithelial cells were morphologically normal and tightly arranged, and lymphocyte punctate infiltration was visible in the lamina propria (black arrow); while the epithelial structure of the positive drug group was intact, the epithelial cells were morphologically normal and tightly arranged, and no obvious inflammation was observed in the lamina propria.

[0086] The above experimental results were matched with the four diagnostic indicators to judge whether the model was successful: the exhaustive swimming time of the rats in the model group was significantly shortened, and their endurance deteriorated, which was consistent with the main symptom of fatigue; at the same time, the rats in the model group had a reduced food intake, and their palm temperature was lower than that of the rats in the normal group, and their soles were pale and bloodless, which were consistent with the secondary symptoms of poor appetite and cold limbs; at the same time, the pulse strength of the rats in the model group was only half of that in the control group, which showed a weak pulse, which was consistent with the pulse condition; at the same time, the tongue of the rats in the model group was thin and white, which was consistent with the tongue condition; it basically met the four diagnostic standards.

[0087] In addition, the model group lost weight, which is basically consistent with the emaciated body shape of patients with clinical spleen and stomach deficiency. At the same time, qi deficiency can lead to poor blood circulation, which can be manifested as blood circulation disorders in the extremities, that is, cold limbs, and symptoms of blood stasis. The experimental results also showed that PT and TT in the coagulation indicators were shortened, which is clinically significant for blood stasis.

[0088] At the same time, Xiangsha Liujunzi Decoction, as a classic prescription for treating spleen and stomach qi deficiency type gastritis, can significantly improve the main symptom indicators of rats after one week of oral administration. Therefore, it is more powerful proof that exhaustive swimming-induced spleen and stomach qi deficiency type gastritis is successful.

[0089] In summary, exhaustive swimming to simulate overwork and oral administration of ammonia to simulate the invasion of Helicobacter pylori can successfully induce rats to show the pathological and syndromic characteristics of spleen and stomach qi deficiency type gastritis for two months.

[0090] The success rate data statistics of the spleen and stomach qi deficiency type gastritis model obtained by the construction method of this embodiment are shown in Table 3:

[0091] Table 3 Data on the success rate of constructing the spleen and stomach qi deficiency type gastritis model

[0092] Detection indicators Number of model groups Number of successful models Success rate weight 12 12 100％ Palm temperature 12 8 66.7％ Rectal temperature 12 8 66.7％ Swimming time to exhaustion 12 10 83.3％ PT / TT 12 9 75％ Gastric tissue pathology 12 8 66.7％ Tongue coating color 12 8 66.7％

[0093] The results in Table 3 show that the success rate of the spleen and stomach qi deficiency type gastritis model obtained by the construction method of the present invention is relatively high, reaching more than 66.7% overall, and the symptoms of liver and stomach disharmony in rats still exist one week after stopping the modeling. If the model needs to be maintained in the future, it only needs to be maintained by exhaustive swimming, and the number of exhaustive swimming can be reduced from 2 times a day to 3 times a week.

[0094] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for constructing an animal model of spleen and stomach qi deficiency type gastritis, characterized in that: It involves inducing the experimental animals to swim at full capacity and to take oral ammonia water instead of their daily drinking water.

2. The construction method according to claim 1, characterized in that: The experimental animals include rats.

3. The construction method according to claim 2, characterized in that: The exhaustive swimming exercise is performed 1 to 3 times a day.

4. The construction method according to claim 3, characterized in that: The concentration of the ammonia water is 0.1-0.3 wt %.

5. The construction method according to claim 4, characterized in that: The induction time is 2-3 months.

6. The construction method according to claim 5, characterized in that: The construction method also includes testing the body weight, food intake and water intake of the experimental animals during the induction period, and testing the rectal temperature and palm temperature, endurance, respiratory amplitude and pulse amplitude, coagulation analysis, and gastric pathology of the experimental animals after the induction.

7. An animal model of spleen and stomach qi deficiency type gastritis obtained by the construction method according to any one of claims 1 to 6.

8. Use of the animal model of spleen-stomach-qi deficiency type gastritis as claimed in claim 7 as an animal model in the research of spleen-stomach-qi deficiency type gastritis.

9. Use of the animal model of spleen-stomach-qi deficiency gastritis as claimed in claim 7 in screening drugs for spleen-stomach-qi deficiency gastritis.

10. Use of the animal model of spleen-stomach-qi deficiency gastritis as claimed in claim 7 in evaluating the effect of drugs on spleen-stomach-qi deficiency gastritis.

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