CN115349498B

CN115349498B - Construction method of damp-heat type endometritis mouse model

Info

Publication number: CN115349498B
Application number: CN202211113443.2A
Authority: CN
Inventors: 李宁; 陈继香; 杨素芳; 赵湘培; 鞠功辰昊
Original assignee: International Zhuang Medical Hospital Of Guangxi
Current assignee: International Zhuang Medical Hospital Of Guangxi
Priority date: 2022-09-14
Filing date: 2022-09-14
Publication date: 2023-07-25
Anticipated expiration: 2042-09-14
Also published as: DE102023119315A1; CN115349498A

Abstract

The invention provides a method for constructing a damp-heat type endometritis mouse model, belonging to the technical field of animal model construction; in the invention, a high-fat high-sugar feed is adopted to feed the mice in a damp-heat environment, and in the feeding process, the mice are matched with single-day oil filling and feeding and double-day white spirit filling and feeding to obtain damp-heat syndrome model mice; and modeling the damp-heat syndrome model mice by using an endometritis disease model to obtain a damp-heat type endometritis mouse model. The construction method fills the blank of the damp-heat type endometritis animal model. Therefore, the experiment is used for modeling of damp-heat syndrome models and disease models, and provides reliable technical support for exploring a mechanism for treating damp-heat type endometritis. The construction method of the invention is divided into damp-heat syndrome model modeling and disease model modeling, wherein the operation modeling of the disease model has high success rate (99%) and stable model compared with the traditional vaginal perfusion Lipopolysaccharide (LPS).

Description

Construction method of damp-heat type endometritis mouse model

Technical Field

The invention belongs to the technical field of animal model construction, and particularly relates to a method for constructing a damp-heat type endometritis mouse model.

Background

Endometritis is a gynecological disease characterized by inflammation of endometrial glands and interstitial tissue, and is classified into acute and chronic diseases, and can be independently developed or accumulated in a plurality of parts. Acute endometritis is less common due to the wide application of antibiotics. Chronic Endometritis (CE) is ignored due to lack of specific clinical symptoms, and researches show that CE can cause a series of adverse effects such as increased female infertility risk, embryo implantation failure, abnormal uterine bleeding and the like.

Ancient books of traditional Chinese medicine have no endometritis record, and according to the symptom characteristics of endometritis, the traditional Chinese medicine belongs to traditional Chinese medicine 'leukorrhagia', 'abdominal pain of ladies', 'heat entering blood room' and the like. Traditional Chinese medicine considers that the pathogenesis of the traditional Chinese medicine is complex, but the traditional Chinese medicine can be summarized into 5 aspects of dampness, heat, blood stasis, cold and deficiency. Damp-heat is the main causative factor of endometritis, and blood stasis is the root cause of the disease. Therefore, it is important to construct a damp-heat type endometritis model and search for a deep therapeutic mechanism.

However, there is no description of constructing an animal model of damp-heat type endometritis.

Disclosure of Invention

The invention aims at a construction method of a damp-heat type endometritis mouse model, and fills the blank of the damp-heat type endometritis animal model.

The invention provides a method for constructing a damp-heat type endometritis mouse model, which comprises the following steps:

1) Feeding the mice with high-fat high-sugar feed under a damp-heat environment, wherein in the feeding process, the mice are matched with single-day filling and feeding of oil and double-day filling and feeding of white spirit, so that damp-heat syndrome model mice are obtained;

the feeding period is 25-30 d;

the raising time in the damp-heat environment is 8-10 h each day;

2) And modeling the damp-heat syndrome model mice by using an endometritis disease model to obtain a damp-heat type endometritis mouse model.

Preferably, the temperature of the damp-heat environment is 29.85-30.15 ℃; the humidity of the damp-heat environment is 80-90%.

Preferably, the high-fat high-sugar feed comprises the following components in percentage by mass: 55% of basic feed, 12% of palm oil, 18% of sucrose, 3% of cholesterol, 7% of yolk powder and 5% of protein powder.

Preferably, the degree of the white spirit is 50-56 degrees.

Preferably, before filling the white spirit, the white spirit is diluted by water, and the volume ratio of the water to the white spirit is (1-1.2): 1; the filling amount of the diluted white spirit is 0.5-0.8 ml/100g of the weight of the mice.

Preferably, the oil is fed in an amount of 0.5 to 0.8ml/100g of mouse body weight.

Preferably, the method for modeling the endometritis disease model of the damp-heat syndrome model mouse comprises the steps of sequentially anesthetizing the damp-heat syndrome model mouse, injecting an LPS aqueous solution into uterine cavity and injecting physiological saline into abdominal cavity; the concentration of LPS in the LPS aqueous solution is 2-2.5 mug/ul; the injection quantity of the LPS aqueous solution is 20-25 mu l/piece; the injection amount of the physiological saline is 0.08-0.1 ml/10g of the weight of the mice.

The invention provides a construction method of a damp-heat type endometritis mouse model, which comprises the steps of feeding a mouse with high-fat and high-sugar feed in a damp-heat environment, and feeding white spirit by filling in a single day and filling in a double day in the feeding process to obtain a damp-heat type endometritis mouse model; and modeling the damp-heat syndrome model mice by using an endometritis disease model to obtain a damp-heat type endometritis mouse model.

The damp evil is the internal and external dampness, the internal dampness is related to improper diet (sweet and greasy diet, insufficient spleen yang, water dampness transformation, and damage to the pulse) and the external dampness is related to the living humid environment. The method for constructing the invention is to put the mice in a damp-heat environment, feed the mice with high-fat and high-sugar feed, and adopt white spirit to promote injury of spleen and stomach, so as to construct a damp-heat syndrome model together; and then modeling the damp-heat syndrome model mice by using an endometritis disease model to obtain a damp-heat type endometritis mouse model. The animal model of traditional Chinese medicine standard should be divided into syndrome and disease model, considering the complex dialectics of syndrome model, it can be modeled by means which can not be understood by ordinary people, and it is difficult to succeed, and in most modeling, multiple disease models are created. In clinic, the traditional Chinese medicine diagnosis of endometritis is mostly damp-heat type, but no modeling method for damp-heat type endometritis exists in the prior art, and the construction method fills the blank of the damp-heat type endometritis animal model. The invention performs the modeling of the damp-heat syndrome model and the modeling of the disease model, and provides reliable technical support for exploring the mechanism for treating damp-heat type endometritis. The construction method of the invention is divided into damp-heat syndrome model modeling and disease model modeling, wherein the operation modeling of the disease model has high success rate (99%) and stable model compared with the traditional vaginal perfusion Lipopolysaccharide (LPS).

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic representation of a surgical procedure with intra-uterine injection of LPS;

FIG. 2 is a schematic diagram of a mouse damp-heat model, wherein A is a normal group, and B is a damp-heat syndrome model group;

FIG. 3 shows uterine morphology changes in mice of each group, wherein A is blank group, B is model group, and C is treatment group;

FIG. 4 shows the uterine HE staining (200X) of each group of mice, with a scale of 100 μm, wherein A is blank, B is model, and C is treatment;

FIG. 5 shows the expression of endometrial CD14 (x 200) in each group of mice, wherein A is the blank group, B is the model group, and C is the treatment group;

FIG. 6 is a graph showing results of comparative statistical analysis of average Optical Density (OD) of mouse endometrium CD14, wherein the model group is compared with the blank group ^## P <0.01, model group compared with treatment group ^＊＊ P＜0.01；

FIG. 7 shows the expression of gastric AQP3 in mice of each group (x 200), wherein A is blank group, B is model group, and C is treatment group;

FIG. 8 is a graph showing the results of a comparative statistical analysis of average optical density (OD values) of mouse stomach AQP3, compared to the blank, model group ^## P <0.01, model group compared with treatment group ^＊＊ P＜0.05；

FIG. 9 shows the results of statistical analysis of the expression of inflammatory factors (pg/ml) in mice of each group, wherein A is TNF-a, B is IL-10, C is IL-8, compared to the blank group, model group ^## P <0.01, model group compared with treatment group ^＊＊ P＜0.05；

FIG. 10 shows HE staining results (200X) for tongue tissue from each group, where A is blank, B is model, and C is treatment.

Detailed Description

the feeding period is 25-30 d;

the raising time in the damp-heat environment is 8-10 h each day;

Firstly, raising a mouse in a damp-heat environment by adopting a high-fat high-sugar feed, wherein in the raising process, the mice are matched with single-day oil filling and feeding and double-day white spirit filling and feeding to obtain damp-heat syndrome model mice; the feeding period is 25-30 d; the daily raising time in the damp-heat environment is 8-10 h, and the daily raising time in the damp-heat environment is preferably 10:00-18:00.

In the invention, the temperature of the humid and hot environment is preferably 29.85-30.15 ℃, more preferably 30 ℃; the humidity of the hot and humid environment is preferably 80% to 90%, more preferably 85%.

In the invention, the alcohol content of the white spirit is preferably 50-56 degrees; the white spirit is preferably 56 DEG red star Erguotou, produced by Beijing red star Co., ltd. In the invention, before filling the white spirit, the white spirit is preferably diluted by water, and the volume ratio of the water to the white spirit is preferably (1-1.2): 1; the filling amount of the diluted white spirit is preferably 0.5-0.8 ml/100g of the weight of the mice.

In the present invention, the feeding amount of the oil is preferably 0.5 to 0.8ml/100g of the weight of the mice; the oil preferably includes vegetable oil and animal oil; the vegetable oil preferably comprises peanut oil.

After the damp-heat syndrome model mice are obtained, the invention carries out endometritis disease model modeling on the damp-heat syndrome model mice to obtain damp-heat type endometritis mouse models.

The method for modeling the endometritis disease model is not particularly limited, and the method for modeling the endometritis disease model is conventional in the art.

In the invention, the method for modeling the endometritis symptom model of the damp-heat symptom model mouse preferably comprises the steps of sequentially anesthetizing the damp-heat symptom model mouse, injecting an LPS aqueous solution into uterine cavity and injecting physiological saline into abdominal cavity; the concentration of LPS in the LPS aqueous solution is preferably 2-2.5 mug/ul; the injection amount of the LPS aqueous solution is preferably 20-25 mu l/L; the injection amount of the physiological saline is preferably 0.08-0.1 ml/10g of the body weight of the mice.

In the present invention, the anesthesia mode is preferably intraperitoneal injection of pentobarbital sodium, and the injection amount of the pentobarbital sodium is preferably 40-70 mg/kg of the body weight of the mouse, and more preferably 50-60 mg/kg of the body weight of the mouse.

Before the aqueous solution of LPS is injected into the uterine cavity, the invention preferably also comprises the steps of skin preparation, disinfection, abdomen placement from the position 0.4 cm to 0.5cm below the kidney region at the back, white ovarian tissue finding out the uterus, and the aqueous solution of LPS is injected into the uterine cavity; the surgical access adopted by the surgery is a back surgical path, so that the chance of contacting with intestinal tracts is avoided, surgical complications such as postoperative infection, intestinal flatulence, intestinal adhesion, intestinal obstruction and the like are reduced, the extra interference caused by molding is reduced, and the success rate of molding is ensured.

In the present invention, the intrauterine injection of aqueous solution of LPS is preferably performed by using a micro-syringe.

After the aqueous solution of LPS is injected into uterine cavity, the invention preferably closes abdomen according to anatomical level, disinfects, injects physiological saline into abdominal cavity to supplement liquid, and reduces hypovolemia, hemorrhagic shock and death caused by fasted and operation.

After the physiological saline is injected into the abdominal cavity, the preferred choice of the invention is that the artificial climate box is placed in the lateral position or the prone position for reviving, so that death caused by hypothermia due to anesthesia is reduced, and choking death caused by tongue root falling and vomiting is avoided; the temperature in the climatic chamber is preferably 28 ℃. The invention simulates the post-operation management mode in clinical operation and ensures the success rate of molding.

The method is suitable for all animal operation modeling.

For further explanation of the present invention, the method for constructing a damp-heat type endometritis mouse model according to the present invention will be described in detail with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.

Example 1

1. Experimental animal

SPF-class 8-10 week Kunming female mice 30, average body weight 30-35 g, purchased from Gekko Biotechnology Co., ltd., animal eligibility No.430726220100069835, produced license number SCXK 2019-0014.

2. Feed stuff

The high-fat and high-sugar feed is processed by animal experiment centers of Guangxi university of Chinese medicine, and the mass ratio of the formula is as follows: 55% of basic feed, 12% of palm oil, 18% of sucrose, 3% of cholesterol, 7% of egg yolk powder and 5% of protein powder; the basic feed is provided by animal experiment centers of university of Chinese medicine in Guangxi province.

3. Test drug

Levofloxacin hydrochloride tablets: zhejiang Beijing New pharmaceutical industry Co., ltd (approval document: national medicine standard H19990060), 0.1 g.times.6 tablets.times.1 plates/box.

4. Experimental reagent

56 ° red star er guotor (produced by beijing red star inc); LPS 10mg (sigma Co., USA); TNF-a ELISA kit (WUHan Bei Yinlai Biotechnology Co., ltd.); protein quantification kit (white shark biotechnology Co., ltd.); CD14 rabbit monoclonal antibody (Beijing Soy Bao technology Co., ltd.); AQP3 monoclonal resistance (beijing soleba technologies limited); xylene national pharmaceutical group chemical company, inc; normal rabbit serum: servicebio; hematoxylin dye liquor: servicebio; hematoxylin blue-returning liquid: servicebio; universal two-step kit: a fir gold bridge;

5. experimental instrument

100 μl of tip micro laboratory sample injection needle is available from Shanghai Gao Ge industry Co., ltd; a dehydrator: DIAPATH Donatello; pathological section machine: leica instruments, shanghai, inc. RM2016; tissue spreading machine: jin Huashi Kedi instruments and equipment limited, zhejiang province, KD-P; immunohistochemical pen: servicebio WG1066-1; and (3) a microscope: nikon E100; imaging system: nikon DS-U3;5430R high speed refrigerated centrifuge: ai Bende China Co., ltd; centrifuge 5810R cryocentrifuge: germany; DW-86-L290-80 ℃ refrigerator: macadamia.

1. The test method comprises the following steps:

1. animals were grouped and after one week of adaptive rearing, 10 blank groups were randomly drawn, and the remaining 20 were modeled with a damp-heat syndrome model. And after the symptom model is judged to be successful, performing operation disorder model modeling, and randomly dividing 20 damp-heat type mice into a model group and a treatment group.

2. Moulding

2.1 symptomatic modeling

Blank group: SPF laboratory room temperature 20-25 deg.C, humidity 50% -60%, ordinary fodder and water feeding for 28 days;

damp-heat model group: the set temperature of the artificial climate box in the damp-heat mode is 30+/-0.15 ℃, the humidity is 80% -90%, the artificial climate box is fed with high-fat and high-sugar feed every day for 8 hours (10:00-18:00), white spirit is filled every day (the volume ratio of 56 DEG white spirit to drinking water is 120:100,0.5ml/100 g), and the weight of mice is 0.5ml/100g when peanut oil is filled every day. The physical changes were measured weekly and the symptom model evaluation was performed 28 days later.

2.2 pharmaceutical intervention

After day 28, the stomach was irrigated, and the blank group and model group were irrigated with equal volumes of physiological saline daily. In the treatment group, levofloxacin hydrochloride tablets are smashed, physiological saline is diluted to 18mg/ml, and the stomach is continuously irrigated for 5 days 1 time per day according to 3.6mg (0.2 ml) of the body weight each time.

2.3 disease modeling

Injecting pentobarbital sodium 40-70 mg/kg into abdominal cavity 1h after last gastric lavage, preparing skin, sterilizing, placing into abdomen about 0.5cm below kidney region from back, finding white ovary tissue, finding uterus along with vigor, injecting LPS (2.5 μg/μl) 25 μl (see figure 1) into uterine cavity of mini-injector, sterilizing, injecting physiological saline 0.1ml/10g into abdominal cavity, and placing into artificial climatic box at 28deg.C for waking up at lateral position or prone position to avoid choking death caused by tongue root falling and emesis. The blank group is not processed. 24 hours later, the eyeballs are taken for blood, and the mice are sacrificed for cervical vertebra removal to collect relevant specimens and measure indexes.

3. Detection index

3.1 uterine pathology detection the uterine tissue of the mice was taken, fixed with 4% formaldehyde, paraffin embedded, sectioned, HE stained and examined under microscope for each group of pathological tissue changes.

3.2 immunohistochemical detection of the expression of gastric AQP3 and uterine CD14

Dyeing by SABC method, dewaxing with xylene, displacing xylene with gradient ethanol to water, and washing with distilled water for 5min. Antigen retrieval was performed for 8min and washing was performed for 5min×3. Incubation is carried out for 25min by 3% hydrogen peroxide, and washing is carried out for 3 times. Blocking with 3% BSA for 30min, primary antibody incubated overnight at 4 ℃. Washing for 3 times, incubating the secondary antibody for 50min, developing DAB, and stopping distilled water. Hematoxylin counterstain, dehydration, transparent and neutral resin sealing, microscopic observation and image acquisition and analysis.

3.3ELISA method for detecting serum TNF-a, IL-10 and IL-8 eyeball blood collection, centrifugation and preservation at-80 ℃, and detection and analysis of the content of mouse serum TNF-a, IL-10 and IL-8 according to ELISA kit steps.

2. Experimental results:

1. mice in the blank group have good mental state, sensitive response, normal drinking water, luster hair, normal urination and defecation, and normal vaginal secretion under the condition of modeling. The rest groups evaluate the damp-heat syndrome and disease model: the mice are listlessness, slow reaction, dull and lusterless hair, reduced drinking water, lips Zhou Anzi, yellow and odorous urine, loose stool, anal Zhou Wuhui and increased vaginal secretion, and the model accords with the syndrome model. See fig. 2.

2. Uterine morphology changes in mice of each group: normal group: the uterus has no congestion and edema and normal morphology; model group uterus: the volume is increased, the lumen is expanded, the serosa surface is obviously swollen, engorged with blood and has dark red color; treatment group: the uterus size, swelling, congestion, etc. are relieved to different extents. See fig. 3.

3. Mice uterine HE staining results blank: the structure of each layer of uterus is clear, the epithelium of endomembrane is complete, the cell morphology is normal, the number of the uterus glands of the lamina propria is rich, the distribution is even, the myoblast arrangement is regular, and no obvious abnormality is seen. Model group: uneven endometrium thickness, necrosis of large amount of epithelial cells of endometrium, deep dyeing, fragmentation or dissolution and disappearance of nucleus; the small amount of the lamina propria of the uterus gland epithelial cells are necrotized, and the nuclei are deeply stained, disintegrated or dissolved and disappear; the intimal lamina propria and the myometrium both see a large amount of granulocyte infiltration (black arrows). Treatment group: the intimal lamina propria and the myometrium are both seen with a small amount of granulocyte infiltration. See fig. 4.

4. Uterine CD14 immunohistochemical results for each group of mice: microscopic observation (tan film staining was strongly positive for CD 14), and the model group was darker brown compared to the normal group, indicating more expression of CD14, and the treatment group was significantly less colored compared to the model group, indicating less expression of CD14 under the action of levofloxacin hydrochloride. See fig. 5. Through gray scale analysis, the CD14 gray scale value in the model group is obviously reduced (P < 0.01); the treatment group had elevated CD14 gray values (P < 0.01) compared to the model group, see fig. 6. The results suggest that CD14 expression and the degree of endometritis are positively correlated, indicating successful modeling of the model of endometritis disorder. Levofloxacin hydrochloride can control the development of uterine tissue inflammation by reducing the expression of CD14 on endometrial tissue cells. CD14 acts as a receptor for LPS, recognizes and binds to LPS, stimulating the secretion of inflammatory factors by monocytes.

5. Gastric AQP3 immunohistochemical results in mice of each group: observed under a microscope, the brown yellow color is positive to AQP3, and the brown color is strong positive. The model group was darker brown compared to the normal group, indicating greater expression of gastric AQP3, and the treatment group had less coloration than the model group, indicating reduced expression of AQP3 in levofloxacin hydrochloride. See fig. 7. Through OD value analysis, compared with a blank group and a model group, the AQP3OD value is remarkably increased (P < 0.01); the AQP3 grey values were significantly reduced (P < 0.01) in the treatment group compared to the model group, see fig. 8. The result shows that the AQP3 is positively correlated with the damp-heat model degree, the damp-heat model is successfully molded, and the levofloxacin hydrochloride has the effects of reducing the AQP3 and treating damp-heat type endometritis. Aquaporins (AQPs) are a family of membrane proteins that are widely found in various human tissue cells and allow water to pass through the biological membrane to maintain in vivo water metabolic balance. The normal expression of AQP3 may be the molecular biological basis for transporting and transforming water dampness and body fluid distribution, and the abnormal expression may be one of the important reasons for the formation of damp evils, which are closely related to the expression level of aquaporin.

6. ELISA assay for release of inflammatory factors from mice of each group: compared with blank group, the expression of inflammatory factors TNF-a and IL-8 in model group is obviously raised, and has statistical significance, P is less than 0.01. The expression of the inflammatory factors TNF-a and IL-8 was significantly reduced (P < 0.05) in the model group compared to the treatment group. The model of endometritis is successfully modeled, and the levofloxacin hydrochloride has obvious effect of inhibiting endometritis. The expression of IL-10 was significantly reduced (P < 0.01) in the model group compared to the red and white group, and the expression of IL-10 was significantly increased (P < 0.01) in the model group compared to the treatment. IL-10 is inversely related to inflammatory expression and has the effect of inhibiting inflammatory response. Further proves that the model modeling of the endometrium inflammation candidate model is successful. See fig. 9.

7. Compared with the blank group tongue tissue, the model group tongue has increased mucosa thickness, increased mucosal layer keratinization, deeper color staining, obvious lamina propria inflammation, congestion and edema, and significantly increased nipple density and frequency of nuclear division. The improvement in lingual mucosa thickness and keratinization was not evident in the treated group compared to the model group, but the inflammatory changes in the lamina propria were improved, see fig. 10.

8. The sense of the research on the proinflammatory cytokines TNF-alpha, IL-8 and anti-inflammatory factor IL-10 is that the TNF-alpha is produced by mononuclear macrophages and has wide biological activity. To perform physiological functions of modulating immune responses and combating infections, TNF- α becomes a serious inflammatory transmitter if it is at a higher concentration in the body. IL-8 is secreted by neutrophils, T cells, and basophils and is an important chemokine that plays an important role in the inflammatory response. IL-10 is produced by Th2 cells, has strong anti-inflammatory effect, and is the main anti-inflammatory factor studied at present. It can directly inhibit macrophage function, inhibit excessive expression of proinflammatory cytokines IL-6, TNF-alpha, etc. and control inflammatory waterfall reaction, and reduce excessive damage of inflammation to organism.

In conclusion, the invention adopts a multi-factor pathogenic damp-heat type endometritis mouse model, and lays a foundation for deep exploration of treatment of damp-heat type endometritis.

Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.

Claims

1. A construction method of a damp-heat type endometritis mouse model comprises the following steps:

1) Feeding the mice with high-fat high-sugar feed under a damp-heat environment, wherein in the feeding process, the mice are matched with single-day filling and feeding of oil and double-day filling and feeding of white spirit to obtain damp-heat syndrome model mice;

the feeding period is 25-30 d;

the raising time in the damp-heat environment is 8-10 h each day;

2) Modeling the damp-heat syndrome model mice by using an endometritis disease model to obtain a damp-heat type endometritis mouse model;

the temperature of the wet and hot environment is 29.85-30.15 ℃.

2. The method of claim 1, wherein the humidity of the hot and humid environment is 80% to 90%.

3. The construction method according to claim 1, wherein the high-fat and high-sugar feed comprises the following components in percentage by mass: 55% of basic feed, 12% of palm oil, 18% of sucrose, 3% of cholesterol, 7% of yolk powder and 5% of protein powder.

4. The method according to claim 1, wherein the degree of white spirit is 50-56 °.

5. The construction method according to claim 4, wherein the white spirit is diluted with water before the filling of the white spirit, and the volume ratio of the water to the white spirit is (1-1.2): 1; the filling amount of the diluted white spirit is 0.5-0.8 ml/100g of the weight of the mice.

6. The construction method according to claim 1, wherein the oil is fed in an amount of 0.5 to 0.8ml/100g of the body weight of the mouse.

7. The method of claim 1, wherein modeling the damp-heat syndrome model mice for endometritis disorder comprises sequentially anesthetizing the damp-heat syndrome model mice, intrauterine injection of an aqueous solution of LPS, and intraperitoneal injection of physiological saline; the concentration of LPS in the LPS aqueous solution is 2-2.5 mug/ul; the injection quantity of the LPS aqueous solution is 20-25 mu l/piece; the injection amount of the physiological saline is 0.08-0.1 ml/10g of the weight of the mice.