CN115349498A

CN115349498A - Construction method of damp-heat type endometritis mouse model

Info

Publication number: CN115349498A
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: 2022-11-18
Anticipated expiration: 2042-09-14
Also published as: CN115349498B; DE102023119315A1

Abstract

The invention provides a construction method of a damp-heat type endometritis mouse model, belonging to the technical field of animal model construction; in the method, a high-fat high-sugar feed is adopted to feed the mice in a damp-heat environment, and in the feeding process, oil feeding is performed in a single-day matched mode, and white spirit feeding is performed in a double-day matched mode, so that the damp-heat syndrome model mice are obtained; and (3) carrying out endometritis disease model modeling on the damp-heat syndrome model mouse to obtain a damp-heat type endometritis mouse model. The construction method of the invention fills the blank of the damp-heat type endometritis animal model. Therefore, the experiment is used for modeling a damp-heat syndrome model and a disease model, and provides reliable technical support for researching 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 vagina pouring 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 construction method of a damp-heat type endometritis mouse model.

Background

Endometritis is a gynecological disease characterized by inflammation of endometrial glands and interstitial tissues, which is divided into acute and chronic diseases, and can be singly attacked or accumulated in a plurality of parts. Due to the widespread use of antibiotics, acute endometritis is relatively rare. Chronic Endometritis (CE) is overlooked due to lack of specific clinical symptoms, and researches show that CE can cause a series of adverse effects such as female infertility risk increase, embryo implantation failure, abnormal uterine bleeding and the like.

The ancient books of traditional Chinese medicine have no record of endometritis, and belong to the morbid leucorrhea, bellyache of husband, heat entering blood room and the like of the traditional Chinese medicine according to the symptom characteristics of endometritis. TCM considers that the etiology is multiple and the pathogenesis is complex, but it can be summarized into 5 aspects of dampness, heat, stasis, cold and deficiency. Damp-heat is a main pathogenic factor of endometritis, and blood stasis blocking is the basic pathogenesis of the endometritis. Therefore, it is important to construct a damp-heat type endometritis model and to deeply explore the treatment mechanism.

However, there is no record on the construction of an animal model of damp-heat endometritis at present.

Disclosure of Invention

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

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

1) Feeding the mice with high-fat high-sugar feed in a damp-heat environment, wherein in the feeding process, oil feeding is performed in a single-day mode, and liquor feeding is performed in a double-day mode to obtain damp-heat syndrome model mice;

the raising period is 25-30 d;

the feeding time in a damp and hot environment is 8-10 h every day;

2) And (3) carrying out endometritis disease model modeling on the damp-heat syndrome model mouse to obtain a damp-heat type endometritis mouse model.

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

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

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

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

Preferably, the feeding amount of the oil is 0.5-0.8 ml/100g of mouse body weight.

Preferably, the step of carrying out endometritis symptom model building on the damp-heat syndrome model mouse comprises the steps of sequentially anaesthetizing the damp-heat syndrome model mouse, injecting an LPS (lipopolysaccharide) aqueous solution into a uterine cavity and injecting physiological saline into an abdominal cavity; the concentration of the LPS in the LPS water solution is 2-2.5 mu g/mu l; the injection amount of the LPS aqueous solution is 20-25 mul/piece; the injection amount of the normal saline is 0.08-0.1 ml/10g of mouse body weight.

The invention provides a construction method of a damp-heat type endometritis mouse model, which comprises the steps of feeding a mouse by adopting high-fat high-sugar feed under a damp-heat environment, and feeding oil on a single day and white spirit on double days in a matching manner in the feeding process to obtain a damp-heat syndrome model mouse; and (3) carrying out endometritis disease model modeling on the damp-heat syndrome model mouse to obtain a damp-heat type endometritis mouse model.

Dampness pathogen, dampness pathogen is internal dampness and external dampness, the internal dampness is related to improper diet (fat, sweet, thick and greasy diet, insufficiency of spleen yang, inability to transport and transform water dampness, injury to collaterals) and the external dampness is related to humid environment of residence. Therefore, the construction method of the invention is that the mice are placed in a damp-heat environment and fed with high-fat and high-sugar feed, and the spleen and the stomach of the mice are promoted to be injured by adopting white spirit to jointly construct a damp-heat syndrome model; and then carrying out endometritis disease model modeling on the damp-heat syndrome model mouse to obtain a damp-heat type endometritis mouse model. The animal model of the traditional Chinese medicine standard should be divided into syndrome and disease models, and considering that the syndrome model is complicated in dialectical form, the model can be made by means which can not be understood by common people, and is difficult to succeed, and most of models are only made into disease models. Clinically, endometritis diagnosed by traditional Chinese medicine is mostly damp-heat type, but a molding method of damp-heat type endometritis does not exist in the prior art, and the construction method fills the blank of a damp-heat type endometritis animal model. The invention carries out model building of damp-heat syndrome model and 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 vagina pouring Lipopolysaccharide (LPS).

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.

FIG. 1 is a view of the modeling of surgical conditions, injected with LPS in the uterine cavity;

FIG. 2 shows the general conditions of the mouse model for damp-heat, wherein A is the normal group and B is the damp-heat syndrome model group;

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

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

FIG. 5 shows the expression of CD14 in endometrium (X200) of each group of mice, wherein A is blank group, B is model group, and C is treatment group;

FIG. 6 is a statistical analysis of the mean Optical Density (OD) of mouse endometrial CD14 compared to a blank model group ^## P is less than 0.01, and the model group is compared with the treatment group ^＊＊ P＜0.01；

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

FIG. 8 is a statistical analysis of the mean optical density (OD values) of mouse gastric AQP3, wherein the model group is compared to the blank group ^## P is less than 0.01, and the model group is compared with the treatment group ^＊＊ P＜0.05；

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

FIG. 10 shows the results of HE staining of tongue tissues in each group (200X) where A is blank, B is model, and C is treatment.

Detailed Description

the feeding period is 25-30 d;

the feeding time in a damp and hot environment is 8-10 h every day;

Firstly, feeding a mouse by adopting high-fat high-sugar feed in a damp-heat environment, wherein in the feeding process, oil feeding is performed in a single-day mode, and liquor feeding is performed in a double-day mode to obtain a damp-heat syndrome model mouse; the feeding period is 25-30 d; the feeding time in the moist heat environment is 8-10 h per day, and the feeding time period in the moist heat environment is preferably 10-00.

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

In the invention, the degree of the white spirit is preferably 50-56 degrees; the white spirit is preferably 56-degree Hongxing Erguotou produced by Beijing Hongxing GmbH. In the invention, before the white spirit is fed, the white spirit is preferably diluted by water, and the volume ratio of the water to the white spirit is preferably (1-1.2) to 1; the feeding amount of the diluted white spirit is preferably 0.5-0.8 ml/100g of mouse body weight.

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

After obtaining the damp-heat syndrome model mouse, the invention carries out endometritis disease model modeling on the damp-heat syndrome model mouse to obtain a damp-heat type endometritis mouse model.

The method for modeling the endometritis disease model is not particularly limited, and a conventional method for modeling the endometritis disease model in the field can be adopted.

In the invention, the model building of the endometritis symptom model of the damp-heat syndrome model mouse preferably comprises the steps of sequentially anaesthetizing the damp-heat syndrome model mouse, injecting LPS aqueous solution into uterine cavity and injecting physiological saline into abdominal cavity; the concentration of the LPS in the LPS water solution is preferably 2-2.5 mu g/mu l; the injection amount of the LPS aqueous solution is preferably 20-25 mu l/piece; the injection amount of the physiological saline is preferably 0.08 to 0.1ml/10g of mouse body weight.

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

Before the LPS aqueous solution is injected into the uterine cavity, the invention preferably also comprises skin preparation, disinfection, placement in the abdomen from the position of 0.4-0.5 cm below the kidney area at the back to see white ovarian tissues, finding out the uterus in a homeopathic manner, and the LPS aqueous solution is injected into the uterine cavity; the operation approach taken by the operation is a back operation path, so that the chance of contacting with intestinal tracts is avoided, the operation complications such as postoperative infection, intestinal flatulence, intestinal adhesion, intestinal obstruction and the like are reduced, the additional interference caused by the model building is reduced, and the success rate of the model building is ensured.

In the present invention, the intrauterine injection of the LPS aqueous solution is preferably performed using a microinjector.

After LPS aqueous solution is injected into the uterine cavity, the abdomen is closed according to the anatomical level, the sterilization is carried out, the normal saline is injected into the abdominal cavity preferentially, and the fluid infusion is carried out, so that the hypovolemia, hemorrhagic shock and death caused by fasting and operation are reduced.

After the normal saline is injected into the abdominal cavity, the preferred lateral decubitus position or prone position is placed into an artificial climate box for reviving, so that death caused by hypothermia due to anesthesia is reduced, and meanwhile, suffocation death caused by tongue root tenesmus and vomiting is avoided; the temperature in the climatic chamber is preferably 28 ℃. The invention simulates the postoperative management mode in clinical operation and ensures the success rate of model making.

The method of the invention is suitable for all animal surgery modeling.

For further illustration of the present invention, the following will describe the construction method of a mouse model of damp-heat endometritis in detail with reference to the drawings and examples, but they should not be construed as limiting the scope of the present invention.

Example 1

1. Laboratory animal

30 Kunming female mice with the SPF grade of 8-10 weeks have the average weight of 30-35 g, are purchased from Tianqin biotechnology limited company of Changsha city, and have the animal qualification number of 430726220100069835 and the production license number of SCXK (Xiang) 2019-0014.

2. Feed stuff

The high-fat high-sugar feed is processed by an animal experiment center of Guangxi Chinese medicinal university, and the formula comprises the following components in percentage by mass: 55% of basal feed, 12% of palm oil, 18% of sucrose, 3% of cholesterol, 7% of yolk powder and 5% of protein powder; the basic feed is provided by animal experiment center of Guangxi Chinese medicine university.

3. Test drugs

Levofloxacin hydrochloride tablets: zhejiang Jing New medicine industries, inc. (approved literature: national drug Standard H19990060), 0.1g × 6 tablets × 1 plate/box.

4. Experimental reagent

56 ° red star Erguotou (manufactured by Beijing red star GmbH); LPS 10mg (sigma corporation, usa); TNF-a ELISA kit (Wuhan Beiyinlie Biotech, inc.); protein quantification kits (white shark Biotechnology Co., ltd.); CD14 rabbit monoclonal antibody (beijing solibao technologies ltd); AQP3 monoclonal antibody (beijing solibao technologies ltd); xylene national chemical group chemical agents ltd; normal rabbit serum: a ServiceBio; hematoxylin staining solution: a ServiceBio; hematoxylin bluing liquid: a ServiceBio; general two-step method kit: china fir gold bridge;

5. laboratory apparatus

A tip micro laboratory sample injection needle of 100 mu l is provided by Shanghai GaoPigeon worker and trade Co., ltd; dewatering machine: DIAPATH Donatello; pathological microtome: shanghai Leica instruments Co., ltd, RM2016; tissue spreading machine: KD-P, kandi instruments and Equipment Co., ltd, jinhua, zhejiang; immunohistochemical pen: servicebio WG1066-1; microscope: nikon E100; an imaging system: nikon DS-U3, japan; 5430R high speed refrigerated centrifuge: ebende China, inc.; centrifuge 5810R low temperature Centrifuge: germany; DW-86-L290-80 ℃ refrigerator: ao De and (5) Koma.

1. The test method comprises the following steps:

1. animal groups mice were bred adaptively for one week, 10 mice were randomly selected as blank groups, and the remaining 20 mice were subjected to damp-heat syndrome model building. After the symptom model is judged to be successful, the model of the operation symptom model is made, and 20 damp-heat mice are randomly divided into a model group and a treatment group.

2. Molding die

2.1 moulding of syndrome model

Blank group: the indoor temperature of an SPF laboratory is 20-25 ℃, the humidity is 50-60%, and the ordinary feed and water are used for feeding for 28 days;

damp-heat model group: the temperature of a damp-heat mode of the artificial climate box is set to be (30 +/-0.15) DEG C, the humidity is 80-90%, the temperature is 8h (10-00-18). Constitutional changes were measured weekly, and symptom model assessment was performed 28 days later.

2.2 pharmaceutical intervention

After 28 days, the stomach was perfused, and the blank group and the model group were perfused with physiological saline of the same volume daily. The treatment group comprises triturating levofloxacin hydrochloride tablets, diluting to 18mg/ml with normal saline, and continuously gavaging for 5 days according to weight 3.6mg (0.2 ml) each time and 1 time/day.

2.3 disease model modeling

1h after last gastric lavage, 40-70 mg/kg pentobarbital sodium is subjected to intraperitoneal injection, skin preparation and disinfection, the medicine is placed into the abdomen from the position which is about 0.5cm slightly below the kidney area behind the back to see white ovarian tissues, the uterus is found out in a proper position, 25 mu l of LPS (2.5 mu g/mu l) is injected into the uterine cavity of a miniature injector (see figure 1), the abdomen is closed according to anatomical levels, the medicine is disinfected, 0.1ml/10g of physiological saline is injected into the abdominal cavity, and the patient is placed into a 28 ℃ artificial climate box to be awakened in a lateral position or a prone position, so that the suffocation death caused by tongue root tenesmus and vomiting is avoided. Blank groups were not processed. After 24h, blood is taken from eyeballs, and the mice are killed after cervical vertebra removal to collect relevant samples and measure indexes.

3. Detecting the index

3.1 uterine pathological examination mouse uterine tissue is fixed by 4% formaldehyde, and pathological tissue changes of each group are observed under a microscope after paraffin embedding, slicing and HE staining.

3.2 immunohistochemical method for detecting the expression of gastric AQP3 and uterine CD14

Dyeing by adopting an SABC method, dewaxing by using dimethylbenzene, replacing the dimethylbenzene by gradient ethanol until water is reached, and washing for 5min by using distilled water. Antigen retrieval was 8min, washing 5min × 3. Incubating with 3% hydrogen peroxide for 25min, and washing for 3 times. 3% BSA blocking for 30min, primary antibody 4 ℃ overnight incubation. Washing for 3 times, incubating the second antibody for 50min, DAB developing, and stopping distilled water. And (4) counterstaining with hematoxylin, dehydrating, transparent, sealing with neutral gum, observing under a microscope, and collecting and analyzing images.

3.3ELISA method for detecting serum TNF-a, IL-10, IL-8 eyeball blood sampling, centrifuging and storing at-80 ℃, and detecting and analyzing the content of mouse serum TNF-a, IL-10, IL-8 according to the steps of ELISA kit.

2. The experimental results are as follows:

1. the mice in the blank group have good mental state, sensitive response, normal drinking water, glossy hair, normal defecation and urination and normal vaginal secretion. The other groups evaluate the damp-heat syndrome model and the disease model: mice are listless and have hypokinesia, slow reaction, dull and lusterless hair, little drinking water amount, dark purple lip, yellow and smelly urine, loose stool, perianal filthy, and vaginal secretion increase, thus conforming to the syndrome model. See fig. 2.

2. Uterine morphology changes in each group of mice: normal group: the uterus has no hyperemia and edema, and the shape is normal; model group uterus: the volume is increased, the lumen is expanded, and the serosal surface is obviously swollen, congested and dark red; treatment groups: the uterus is relieved in size, swelling, congestion and the like. See fig. 3.

3. Mouse uterine HE staining results blank group: the structure of each layer of uterus is clear, the endometrial epithelium is complete, the cell morphology is normal, the number of the uterine glands in the inherent layer is rich, the distribution is uniform, the arrangement of muscle cells in the muscle layer is regular, and no obvious abnormality is seen. Model group: the thickness of the endometrium is uneven, a large amount of epithelial cells of the endometrium are necrotic, and the cell nucleus is deeply dyed, cracked or dissolved and disappears; a small amount of uterine gland epithelial cells in the lamina propria are necrotic, and the cell nucleus is deeply contracted and deeply dyed, cracked or dissolved and disappears; both intimal lamina propria and muscular layer were seen to be infiltrated with a large number of granulocytes (black arrows). Treatment groups: small infiltration of granulocytes was seen in both intimal lamina propria and muscular lamina. See fig. 4.

4. Results of CD14 immunohistochemistry of uterus of mice in each group: and (4) observing under a microscope (the brown membrane is dyed to be strong positive for CD 14), comparing with the normal group, the brown of the model group is darker, which shows that the expression level of CD14 is more, and comparing with the model group, the coloring of the treatment group is obviously lightened, which shows that the expression of CD14 is reduced under the action of levofloxacin hydrochloride. See fig. 5. Through gray level analysis, the CD14 gray level value in the model group is obviously reduced (P < 0.01); the CD14 gray values were elevated in the treatment group compared to the model group (P < 0.01), see fig. 6. The result indicates that the expression of CD14 is positively correlated with the degree of endometritis, which indicates that the model of endometritis disease is successfully modeled. Levofloxacin hydrochloride may 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 LPS, and stimulates monocytes to secrete inflammatory factors.

5. The gastric AQP3 immunohistochemical results of each group of mice are as follows: and through microscopic observation, the color of brown yellow is positive to AQP3, and the color of brown is strong positive. Compared with the normal group, the model group has darker brown color, which indicates that the expression level of gastric AQP3 is more, and compared with the model group, the coloring of the treatment group is reduced, which indicates that the expression of AQP3 can be reduced by levofloxacin hydrochloride. See fig. 7. Through OD value analysis, compared with a model group, the blank group has a remarkably increased AQP3OD value (P < 0.01); the AQP3 gray level was significantly decreased in the treatment group compared to the model group (P < 0.01), see fig. 8. The results show that AQP3 is in positive correlation with the damp-heat model degree, the damp-heat model is successfully molded, and the levofloxacin hydrochloride has the effects of reducing AQP3 and treating damp-heat type endometritis. Aquaporins (AQPs) are a family of membrane proteins that are widely present in various human tissue cells and that pass water through biological membranes to maintain water metabolism balance in the body. The normal expression of AQP3 may be the molecular biological basis for transporting and resolving water dampness and distributing body fluid, and the expression abnormality may be one of the important causes for the formation of damp pathogen, which is closely related to the expression level of aquaporin.

6. ELISA test results of inflammatory factor release of mice in each group: compared with the blank group, the expression of inflammatory factors TNF-a and IL-8 in the model group is obviously increased, and the statistical significance is realized, and P is less than 0.01. Compared with the treatment group, the expression of inflammatory factors TNF-a and IL-8 is obviously reduced in the model group (P < 0.05). The success of the model building of the endometritis model is shown, and the levofloxacin hydrochloride has the obvious effect of inhibiting the endometritis. Compared with the red and white group, the expression of IL-10 in the model group is obviously reduced (P is less than 0.01), and compared with the treatment in the model group, the expression of IL-10 is obviously increased (P is less than 0.01). IL-10 is negatively associated with inflammatory expression and has the effect of inhibiting inflammatory responses. Further proves that the endometritis syndrome model is successfully molded. See fig. 9.

7. Compared with the blank tongue tissue, the model group has the advantages of increased tongue mucosa thickness, increased mucosal layer cornification, darker color dyeing, obvious inflammation, congestion and edema of the inherent layer, and obviously increased nipple density and mitotic phase frequency. The improvement in tongue mucosa thickness and keratosis was not evident in the treated group compared to the model group, but was improved by the alteration of lamina propria inflammation, see figure 10.

8. The research significance of the proinflammatory cytokines TNF-alpha, IL-8 and the anti-inflammatory factor IL-10 is that TNF-alpha is generated by mononuclear macrophage and has wide biological activity. To exert physiological functions of regulating immune response and resisting infection, TNF- α, if present at high concentrations in the body, becomes a serious mediator of inflammation. IL-8 is secreted by neutrophils, T cells, and basophils, is an important chemokine, and plays an important role in inflammatory responses. IL-10 is produced by Th2 cells, has a strong anti-inflammatory effect, and is the main anti-inflammatory factor studied at present. It can directly inhibit the function of macrophage, and also can reduce the excessive damage of inflammation to organism by inhibiting the over-expression of proinflammatory cytokines IL-6, TNF-alpha and the like and controlling the inflammatory waterfall reaction.

In conclusion, the invention adopts a multifactorial pathogenic damp-heat type endometritis mouse model, and lays a foundation for deeply exploring the treatment of damp-heat type endometritis.

Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.

Claims

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

the feeding period is 25-30 d;

the feeding time in a damp and hot environment is 8-10 h every day;

2. The construction method according to claim 1, wherein the temperature of the humid hot environment is 29.85-30.15 ℃; the humidity of the damp and hot environment is 80-90%.

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

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

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

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

7. The construction method according to claim 1, wherein the performing of the endometritis disorder model modeling on the damp-heat syndrome model mouse comprises sequentially anesthetizing the damp-heat syndrome model mouse, injecting an LPS aqueous solution into the uterine cavity, and injecting a physiological saline into the abdominal cavity; the concentration of LPS in the LPS aqueous solution is 2-2.5 mug/mul; the injection amount of the LPS aqueous solution is 20-25 mul/piece; the injection amount of the normal saline is 0.08-0.1 ml/10g of mouse body weight.