Preparation method of silicosis mouse models

A mouse model and silicosis technology, applied in the experimental field, can solve the problems of instillation not in line with the breathing rhythm of mice, good repeatability of mouse silicosis model, increase of external factors of silicosis, etc., to achieve repeatability and High safety, simple and efficient technical methods, and the effect of avoiding physical and mental trauma

Active Publication Date: 2019-11-22
ANHUI UNIV OF SCI & TECH
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Problems solved by technology

[0004] In the past, the preparation of mouse silicosis models was mainly through surgical invasive methods, the risk of infection was difficult to control, and deep injection anesthesia was required for mice, the mortality rate of mice was high, and the repeatability of experiments was poor
In recent years, some researchers have prepared by intratracheal instillation, but the silicosis model produced by this method has the following disadvantages: ① Airway invasion will cause damage to the airway of mice, increasing the external factors that induce silicosis; ②The instillation in the airway does not conform to the breathing rhythm of the mice, and the mouse’s respiratory compliance will be poor during the operation, which increases the risk of death; ③The airway instillation has the possibility of entering the esophagus by mistake, which increases the failure rate of the experiment
So far, there is no mouse model of silicosis with safe operation, good reproducibility, and precise control of injury and fibrosis outcome

Method used

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  • Preparation method of silicosis mouse models

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Experimental program
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Effect test

Embodiment 1

[0021] Example 1. Experimental preparation before mouse modeling

[0022] 1. Adaptation of mice to the environment

[0023] Prepare a systematic system for feeding and experimentation, purchase mice, adapt to the environment for a week, completely randomly divide them into experimental groups and control groups, mark them well, age 7-8 weeks, weigh 18-23g, etc., prepare mice for quantitative anesthesia Machine, high-precision electronic scales, 200ul pipette, preparation of sterile SiO2 suspension, isoflurane, sterile saline, etc.

[0024] 2. Baoding the mouse after anesthesia

[0025] The mice were taken out from the normal temperature animal breeding room, and the basic body weight was recorded. The mice in the model group were anesthetized with isoflurane in an anesthesia machine. When the mice changed from shallow and fast breathing to deep and slow breathing for 3 seconds, they were taken out and placed in the palm of their hands, and nasal drops were performed. Before ...

Embodiment 2

[0026] Embodiment 2. Nasally inhaled SiO2 Preparation of mouse silicosis model

[0027] At the beginning of the nasal drop operation, mix the SiO2 suspension thoroughly and inhale 50-80 μl with a pipette. The tip of the pipette is aimed at the side of the nose of the mouse instead of the tip of the nose, and gently drip a drop. Instill the next drop until the end of the drop, pay attention to observe the breathing state of the mouse while dripping, massage and stroke the chest after the drop is complete, ensure that the vital signs are restored, put it in the cage for observation, and send it back to room temperature after all the experiments are over Breeding in an animal laboratory; put the thumb tip against the jaw of the mouse, and use a micropipette to drip the suspension of SiO2 micron-sized particles through the nose. Every time the mouse inhales a drop, the operator drops a drop until all the drops are finished. Massage externally for 2-3 minutes and put them in the ca...

Embodiment 3

[0028] Example 3. Evaluation of inhalation distribution by color development after adding Evans blue

[0029] Evans blue is an azo dye, which has been used as a chromogenic agent for simulating tracer SiO2 in mouse silicosis modeling (for specific methods, please refer to the literature: Intratracheal instillation and nebulization of bleomycin to induce small A comparative study of a murine model of pulmonary fibrosis). Here we apply Evans blue to the preparation and evaluation stage of a new mouse model of silicosis. After preparing a suitable concentration of dye, we drop different volumes of Evans blue solution into the mice, and after 45 minutes They were anesthetized and killed, and their lungs were fixed in 10% neutral formaldehyde. They were roughly placed and disassembled and placed in a well-lit place to conduct statistical analysis on the distribution of the dye solution and the coloring of each lung lobe.

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Abstract

The invention relates to a preparation method of silicosis mouse models. The preparation method of the silicosis mouse models is characterized in that after experiment preparation, isoflurane in an anesthesia machine is adopted to anesthetize mice in a model group, 3 seconds after rapid-shallow breaths of the mice become deep and slow, each mouse is taken out and placed at the center of a palm, the tip of a thumb presses against the underjaw of each mouse, a micropipette is adopted to instill a SiO2 micro-sized particle suspension through the nose of each mouse, the SiO2 micro-sized particle suspension is instilled into the nose wings to be absorbed, closed-chest massage is conducted for 2-3 minutes, then the mice are placed into cages, and after an experiment is finished, the mice are sent back to a normal-temperature animal feed room; only normal saline of the same volume is used for conducting nasal inhalation on mice in a contrast group; weight indexes of the mice are obtained before and after the experiment, the mice are killed at the first week, the second week, the fourth week, the eighth week and the sixteenth week respectively, lung tissue is taken to be pathologically andbiochemically compared and identified, and a model preparation effect is evaluated. According to the preparation method of the silicosis mouse models, a model production technology is stable and efficient; the method can be repeated many times and is noninvasive, environmentally friendly and humanistic, and labor and time are saved.

Description

technical field [0001] The invention relates to the field of experiment technology, in particular to a method for preparing a mouse model of silicosis. Background technique [0002] Silicosis is an irreversible idiopathic pulmonary fibrosis occupational disease caused by long-term exposure to dust environment. In recent years, ceramics, building materials, mining and other industries have adopted various measures to reduce dust and preventive measures, but silicosis still occurs among occupational groups. In the field of modern energy and safety, the prevention and clinical treatment of silica dust occupational diseases, there are still many major clinical difficult problems that need to be explored and solved urgently. It is urgent to strengthen relevant basic research, such as: the pathological change mechanism of silicosis, the treatment of silicosis and the prevention of silicosis Wait. [0003] In the process of clinical and basic research, issues such as ethics and r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02A01K2207/20A01K2207/30A01K2227/105A01K2267/03
Inventor 李兵刘强陶欣荣穆敏王建华
Owner ANHUI UNIV OF SCI & TECH
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