阅读说明：本技术 一种胆汁反流性胃损伤动物模型的建模方法 (Modeling method of bile reflux gastric injury animal model ) 是由 韩小东 黄勇攀 张炯怡 于 2019-09-24 设计创作，主要内容包括：本发明提供一种胆汁反流性胃损伤动物模型的建模方法,包括以下步骤：步骤1：对实验小鼠其禁食8～15小时、禁饮3～6小时,并保持其活性；步骤2：使用1％～8％的水合氯醛,注射剂量为0.1ml～1.0ml/100g,实验小鼠进行腹腔注射；步骤3：将实验小鼠仰卧位固定,上腹部脱毛,剑突下前正中线切口1～3cm,打开腹腔,暴露肠管,用镊子轻提右侧胃,使其高于皮肤,找到幽门狭窄处,并在狭窄处沿肠管走行切0.5～1.0cm的造口,完全离断幽门括约肌,纵向吻合切口,然后对切口进行缝合消毒；步骤4：对实验小鼠以正常喂养,自由饮水20～40天后得到胆汁反流性胃损伤动物模型。本发明手术创伤小,动物死亡率低,提高了模拟人体胆汁反流性胃损伤发病机制的准确性。(The invention provides a modeling method of a bile reflux gastric injury animal model, which comprises the following steps: step 1: fasting the experimental mouse for 8-15 hours and drinking prohibition for 3-6 hours, and keeping the activity of the experimental mouse; step 2: 1 to 8 percent of chloral hydrate is used, the injection dosage is 0.1 to 1.0ml/100g, and the experimental mouse is injected into the abdominal cavity; and step 3: fixing the experimental mouse in a supine position, unhairing the upper abdomen, cutting a 1-3 cm incision on the front median line of the xiphoid process, opening the abdominal cavity, exposing an intestinal canal, slightly lifting the right stomach with forceps to enable the right stomach to be higher than the skin, finding a pyloric stenosis, cutting a 0.5-1.0 cm stoma at the stenosis along the intestinal canal, completely disconnecting the pyloric sphincter, longitudinally anastomosing the incision, and then suturing and disinfecting the incision; and 4, step 4: the experimental mice are fed normally and are drunk freely for 20-40 days to obtain the bile reflux gastric injury animal model. The invention has small operation wound and low animal mortality, and improves the accuracy of simulating human bile reflux gastric injury pathogenesis.)

1. A modeling method of a bile reflux gastric injury animal model is characterized by comprising the following steps:

step 1: taking a proper experimental mouse, fasting the experimental mouse for 8-15 hours, and keeping the activity of the experimental mouse after drinking for 3-6 hours;

step 2: using 1-8% chloral hydrate, and injecting the injection dose of 0.1-1 ml/100g, and carrying out intraperitoneal injection on the experimental mouse in the step 1;

and step 3: fixing the experimental mouse in the step 2 in a supine position, unhairing the upper abdomen, cutting a 1-3 cm incision on the anterior midline of the xiphoid process, opening the abdominal cavity, exposing an intestinal canal, slightly lifting the right stomach with forceps to be higher than the skin, finding a pyloric stenosis, cutting a 0.5-1 cm stoma along the intestinal canal at the stenosis, completely separating the pyloric sphincter, longitudinally anastomosing the incision, and then suturing and disinfecting the incision;

and 4, step 4: and (4) feeding the experimental mice in the step (3) normally, and freely drinking water for 20-40 days to obtain the bile reflux gastric injury animal model.

2. The method of claim 1, wherein in step 1, the experimental mouse has a body weight of 150-250 g, and is free of gender, the fasting time is 12 hours and the drinking prohibition time is 4 hours.

3. The modeling method of the bile reflux gastric injury animal model according to claim 1, wherein in step 2, 1% -8% chloral hydrate is adopted, and the injection dosage is 0.4 ml-1 ml/100g for intraperitoneal injection to the experimental mouse.

4. The method of claim 3, wherein in step 2, 5% chloral hydrate is used to inject the test mouse into the abdominal cavity at a dose of 0.6ml/100 g.

5. The method for modeling an animal model with bile regurgitation type gastric injury as claimed in claim 1 wherein in step 3, the incision of the inferior-anterior median line of the xiphoid process is 1cm and a 0.5cm stoma is cut along the intestine at the stenosis.

6. The method for modeling an animal model with bile regurgitation type gastric lesion of claim 1 wherein in step 3, the longitudinal anastomotic incision is made by docking the proximal intestinal wall with the distal intestinal wall.

7. The method for modeling an animal model with bile reflux gastric injury as claimed in claim 1, wherein in step 4, the experimental mice are subjected to daily examination and data recording after 15 days.

Technical Field

The invention belongs to the technical field of medical experiments, and particularly relates to a modeling method of a bile reflux gastric injury animal model.

Background

Bile reflux can lead to ulceration, inflammation of the digestive tract and tumours, that is to say chronic bile reflux can lead to hyperplasia, active inflammation, oesophagitis, erosion of the gastric mucosa, gastric ulceration and even the onset of gastric cancer. Has important relation with the occurrence of ulcer, inflammation of upper gastrointestinal tract and tumor;

patients with bile regurgitation stomach injury have stomach fullness or discomfort, which is aggravated after meals, stomachache, or cold stomach, and can be accompanied by abdominal distention, belch, acid regurgitation, heartburn, nausea, emesis, gastric water-shaking sound, borborborygmus, dyschezia, anorexia, emaciation, etc.; severe gastric bleeding, manifested as hematemesis or dark stool (tar-like stool) and positive occult blood test;

the current molding method for chronic gastritis and gastric ulcer comprises the following steps: acid preparation induced acute gastritis model, bile, deoxycholic acid, ammonia water gavage and other gastritis model; stress gastric ulcer model, pyloric ligation method, and ethanol induction model. However, the existing molding method generally has the defects that the operation of an animal model is complex and the death rate is high; and the animal model is not ideal enough for the reflection of clinical indexes and pathological characteristics.

Disclosure of Invention

The invention aims to provide a modeling method of a bile reflux gastric injury animal model, which solves the problems that the operation of the animal model is complicated and the death rate is high in the existing modeling method; and the animal model is not ideal enough for the reflection of clinical indexes and pathological characteristics.

The technical scheme adopted by the invention is that,

the invention provides a modeling method of a bile reflux gastric injury animal model, which comprises the following steps:

step 1: taking a proper experimental mouse, fasting the experimental mouse for 8-15 hours, and keeping the activity of the experimental mouse after drinking for 3-6 hours;

step 2: using 1-8% chloral hydrate, and injecting the injection dose of 0.1-1.0 ml/100g, and carrying out intraperitoneal injection on the experimental mouse in the step 1;

and step 3: fixing the experimental mouse in the step 2 in a supine position, unhairing the upper abdomen, cutting a 1-3 cm incision on the anterior midline of the xiphoid process, opening the abdominal cavity, exposing an intestinal canal, slightly lifting the right stomach with forceps to be higher than the skin, finding a pyloric stenosis, cutting a 0.5-1.0 cm stoma at the stenosis along the intestinal canal, completely separating the pyloric sphincter, longitudinally anastomosing the incision, and then suturing and disinfecting the incision;

and 4, step 4: and (4) feeding the experimental mice in the step (3) normally, and freely drinking water for 20-40 days to obtain the bile reflux gastric injury animal model.

The present invention is also characterized in that,

the weight of the experimental mouse is 150-250 g, the male and female are not restricted, the fasting time is 12 hours, and the drinking prohibition time is 4 hours.

1 to 8 percent of chloral hydrate is adopted, and the injection dosage is 0.4 to 1.0ml/100g to carry out intraperitoneal injection on the experimental mice.

5% chloral hydrate is adopted, the injection dosage is 0.6ml/100g, and the intraperitoneal injection is carried out on experimental mice.

The incision of the anterior midline of the xiphoid process is 1cm, and a 0.5cm stoma is cut at the narrow part along the intestinal canal.

In step 3, the longitudinal anastomosis incision is specifically realized by butting the incision proximal intestinal wall with the incision distal intestinal wall.

In step 4, daily examination and data recording were performed on experimental mice 15 days after entry.

The bile reflux gastric injury animal model has the beneficial effects that the bile reflux gastric injury animal model can be successfully made by simulating the pathogenesis and the reason of gastritis and gastric ulcer and utilizing a new operation mode, so that the bile reflux gastric injury animal model can accurately reflect clinical indexes and pathological characteristics, has high incidence rate of gastric injury, good repeatability and low cost, and has low animal mortality.

Drawings

FIG. 1 is a schematic diagram of the stomach pylorus incised along the long axis of the intestine and the muscular layer of the intestine in the modeling method of the bile reflux gastric injury animal model of the invention;

FIG. 2 is a schematic diagram of a stomach pylorus cut along the long axis of the intestine in the modeling method of the bile reflux gastric injury animal model of the present invention;

FIG. 3 is a schematic diagram of the butt-joint suture of the incision proximal intestinal wall edge and the incision distal intestinal wall distal edge in the modeling method of the bile reflux gastric injury animal model of the invention.

In the figure, 1, stomach, 2, duodenum, 3, lumen of pylorus, 4, sphincter of pylorus, 5, submucosa of pylorus, 6, annular muscle, 7, longitudinal muscle, 8, mucosa, 9, serosa.

Detailed Description

The present invention will be described in detail with reference to the following embodiments.

The invention provides a modeling method of a bile reflux gastric injury animal model, which comprises the following steps:

step 1: taking a proper experimental mouse, fasting the experimental mouse for 8-15 hours, and keeping the activity of the experimental mouse after drinking for 3-6 hours;

step 2: using 1-8% chloral hydrate, and injecting the injection dose of 0.1-1.0 ml/100g, and carrying out intraperitoneal injection on the experimental mouse in the step 1;

and step 3: fixing the experimental mouse in the step 2 in a supine position, unhairing the upper abdomen, cutting a 1-3 cm incision on the anterior midline of the xiphoid process, opening the abdominal cavity, exposing an intestinal canal, slightly lifting the right stomach with forceps to be higher than the skin, finding a pyloric stenosis, cutting a 0.5-1.0 cm stoma at the stenosis along the intestinal canal, completely separating the pyloric sphincter, longitudinally anastomosing the incision, and then suturing and disinfecting the incision;

and 4, step 4: and (4) feeding the experimental mice in the step (3) normally, and freely drinking water for 20-40 days to obtain the bile reflux gastric injury animal model.

The weight of the experimental mouse is 150-250 g, the male and female are not restricted, the fasting time is 12 hours, and the drinking prohibition time is 4 hours.

1 to 8 percent of chloral hydrate is adopted, and the injection dosage is 0.4 to 1.0ml/100g to carry out intraperitoneal injection on the experimental mice.

5% chloral hydrate is adopted, the injection dosage is 0.6ml/100g, and the intraperitoneal injection is carried out on experimental mice.

The incision of the anterior midline of the xiphoid process is 1cm, and a 0.5cm stoma is cut at the narrow part along the intestinal canal.

In step 3, the longitudinal anastomosis incision is specifically realized by butting the incision proximal intestinal wall with the incision distal intestinal wall.

In step 4, daily examination and data recording were performed on experimental mice 15 days after entry.