动物模型,子宫内膜异位症 z-bo 2020-08-05 534

　　Endometriosis (EMT) refers to a disease in which endometrial tissue with the ability to grow appears in other parts of the body other than the mucosa covered by the sub-canal cavity. Clinically, it is typical of abdominal pain, dysmenorrhea, sexual discomfort, and infertility. Symptoms belong to the categories of "dysmenorrhea" and "reverse menstruation" in traditional Chinese medicine. Qi stagnation and blood stasis and kidney deficiency and blood stasis are the main syndrome types.

　　【Modeling mechanism】Based on the establishment of endometriosis disease model by autologous endometrium transplantation, multi-factor intervention methods such as drugs and emotional stimulation are used to create Qi stagnation and blood stasis syndrome model.

　　[Modeling method] Experimental animals: Sprague Dawlev female mature immature rats.

　　1. EMT model 4% chloral hydrate (30ml/kg, intraperitoneal injection) anesthetize the rat, select about 1cm of the upper end of the urethra 1:3 as the entry point, cut 2cm along the midline of the abdomen, separate the abdominal wall layer by layer, enter the pelvic cavity, and explore the rat Uterus (V-shaped). After ligating the local uterine blood vessels with a thin silk thread, take 1～2cm of the left uterine horn and put it in PBS solution immediately, trim and remove the fat tissue outside the uterine serous membrane, cut the uterine cavity longitudinally, and cut 5mm× Make a 5mm uterine film with its endometrial surface opposite to the right peritoneum, and suture its four corners to the abdominal wall with 5-0 silk thread. The remaining part of the uterine film was histologically observed to confirm that the graft was indeed endometrial tissue. After hemostasis, 160,000 U/kg ampicillin dissolved in 0.9% normal saline was dripped into the abdominal cavity to disinfect the abdominal cavity, and the abdominal cavity was closed by suture layer by layer.

　　2. For the model of Qi stagnation and blood stasis syndrome, the rats were given diethylstilbestrol 0.02 mg/kg by gavage on the 5th day before the operation and the 10th day after the operation for 5 consecutive days to keep the rats in the estrus during the operation; intramuscular injection of ampicillin was given after the operation l6 million U/kg for 5 consecutive days. One week after the operation, after the abdominal incision 1:3 is basically healed, the following syndrome interventions are carried out: ①Clamp the tail of the rat with gauze hemostatic forceps to keep it in a state of anger and fighting, 2 minutes/time, 1 time/week; ②Use A thin bandage restrains the hind limbs of the rat, making it difficult to walk, 30 minutes/time, once a week; ③Gavage 1ml/100g of edible oil, 1 time/week; ④Subcutaneous injection of 1g/L adrenaline, 0.2ml/mouse, 1 Times/week. The above methods are performed alternately within 1 week. Observation indicators: rat activity status, mental status, fur, tongue quality, etc., endometriotic lesion growth, pathological and serological indicators, etc.

　　[Model Features] Rats showed symptoms such as listlessness, irritability, curling up, welting, drooping beard, towering and dull coat, red and purple ears, dark purple tongue, and red and purple tails. The disease model and the combined disease and syndrome model have flat epithelial cells in the ectopic endometrium, thin muscle layer and unclear structure. Serum nitric oxide (NO) and endothelin (ET) monoamine neurotransmitters increased, and there was no significant change in serotonin (5-HT).

　　[Model Evaluation and Application] ①The main method of establishing EMT model by surgical transplantation, rabbits, rats, monkeys, etc. can be used for modeling animals, and the methods are the same. ②It is more economical to use rats as model animals and has a high survival rate. ③Addition of epinephrine subcutaneous injection is to simulate the pathophysiological state of the animal during irritation and the etiology of traditional Chinese medicine. After multiple applications, it will cause neuroendocrine dysfunction and abnormal immune function in the animal under stress, and it cannot inhibit or eliminate the ectopic endometrium. Promoting its growth and reproduction is more in line with the pathogenic factors of the disease. ④This model does not require complicated and expensive equipment, as long as the operation method is mastered, it can be copied in a general laboratory (sterile operation). There are many methods for establishing EMT rat models. Among them, the auto-internal membrane transplantation method imitates the cause of human EMT, and has the characteristics of high model stability and high modeling rate. In recent years, it has been widely used in experimental research.