Improving method for preparing mouse silicosis model

A mouse and model technology, applied in animal husbandry and other directions, can solve problems such as failure to successfully prepare mouse models, inability to be widely used, and long modeling time, to ensure repeatability and scientificity, ensure feasibility, The effect of high model success rate

Active Publication Date: 2019-12-31
NORTH CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

SiO2 nebulized daily with the dust system 2 Dust, this method can best simulate the pathogenic process of human dust exposure, which is more in line with the natural progression of silicosis, and has been successfully used in the preparation of rat models; however, this method is limited by special equipment conditions, dust costs and manufacturing costs. Due to the limitations of factors such as long mold time, it cannot be widely used
More importantly, domestic and foreign and the preliminary research of the present invention use micron-sized SiO 2 Dust-infected mice (C57BL / 6 mice were exposed to dust for 6 months, ICR mice were exposed to dust for 2 months) and nanoscale SiO 2 Dust-exposed mice (both C57BL / 6 mice and ICR mice were exposed to dust for 2 months), the results showed that daily nebulized SiO inhalation using the dust-exposed system 2 Dust, SiO in different particle sizes 2 There are very few deposits in the lungs of different strains of mice, and the mouse model cannot be successfully prepared

Method used

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  • Improving method for preparing mouse silicosis model
  • Improving method for preparing mouse silicosis model
  • Improving method for preparing mouse silicosis model

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Improved method for preparing mouse silicosis model

[0041] 1. Experimental animals

[0042] SPF C57BL / 6 mice (provided by the Experimental Animal Center of North China University of Technology), both male and female, 8-16 weeks of age, 20-30g weight.

[0043] 2. Preparation before model preparation

[0044] (1) The micron SiO 2 The granules are ground in an agate mortar for 2-3 hours, and then baked in a vacuum drying oven at 200°C for 2 hours to sterilize and remove endotoxin contamination; use a 0.9% NaCl solution to prepare them to a concentration of 50mg / ml, 100mg / ml, 200mg / ml SiO 2 Suspension.

[0045] (2) The mice were fasted with water for 8h before operation.

[0046] 3. Model preparation process

[0047] (1) Anesthesia: Anesthetize mice by intraperitoneal injection of 1% pentobarbital sodium (50 mg / kg pentobarbital sodium).

[0048] (2) Fixation and shearing: fix the mouse in the supine position, with the neck slightly raised, and the incisor teeth fixed with silk thread...

Embodiment 2

[0056] The mouse model of silicosis prepared by the improved method disclosed in this application is grouped by instilling different concentrations of SiO into the trachea 2 Suspension, specific: Group A injected 50mg / mLSiO 2 Suspension, group B injected 100mg / mLSiO 2 Suspension, group C injected 200mg / mL SiO 2 Suspension, another control group was set up to observe the success rate and mortality of the 4 groups of mouse models, as shown in Table 1.

[0057] Table 1 The success rate and mortality of mouse models

[0058]

[0059] Note: 1. The total number of deaths is equal to the number of deaths during the operation plus the number of deaths after the operation; 2.171 / 223 represents the number of survivals of the model and the total number of survivals (including the number of survivals in the control group and the number of survival in the model group), indicating: 171=60+57 +55, 223=51+60+57+55; 3. The success rate is equal to the number of models divided by the number of surviv...

Embodiment 3

[0065] Compare the number of nodules between animal models prepared by different methods and the mouse model prepared by the improved method of this application (nodules are one of the criteria for judging silicosis)

[0066] 1. References: Quan Shangkun, etc., the intervention effect of Pabirestat on silicosis fibrosis in mice, nasal instillation of SiO 2 The mouse model was prepared by the suspension method, and the nodules of lung tissue sections were observed, such as Picture 10 Shown

[0067] 2. References: Sachin K et al. Oropharyngeal administration of silica in Swiss mice: A robust and reproducible model of occupational pulmonary fibrosis, inhalation of SiO by oropharyngeal 2 The mouse model was prepared by the suspension method, and the nodules of lung tissue sections were observed, such as Picture 11 Shown.

[0068] 3. References: Wu Q et al, miR-489inhibits silica-induced pulmonaryfibrosis by targeting MyD88 and Smad3 and is negatively regulated by lncRNACHRF, inject SiO ...

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Abstract

The invention discloses an improving method for preparing a mouse silicosis model. The improving method includes following steps: step 1, pretreating a mouse, separating jugular anterior fascicles, and exposing trachea; step 2, using smooth forceps to slightly pull the tongue of the mouse out of the mouth, and inserting a 20g blunt needle head into the trachea by about 2-3cm through the front wallof oropharynx under direct view; step 3, using an injector to quickly inject 50microlitres of SiO2 suspension into the trachea, removing the injector, connecting the needle head with a breathing machine, and feeding air for 10-30s: step 4, when normal breathing of the mouse restores, removing the breathing machine, continuing to observe, and pulling out the needle head when breathing is normal; step 5, suturing skin, and performing subsequent nursing and observing. The method is simple and easy in operation, simulates human silicosis process at a high degree and has the advantages of low mouse death rate, high model success rate and high operability.

Description

Technical field [0001] The invention belongs to the technical field of preparation of silicosis animal models, and specifically relates to an improved method for preparing a mouse silicosis model. Background technique [0002] Silicosis is due to long-term inhalation of SiO 2 Dust is a common occupational disease mainly manifested by the formation of silicosis nodules and diffuse pulmonary fibrosis. Understanding the pathogenesis and prevention measures of silicosis depends on the establishment of disease animal models. At present, there are many methods to establish small animal silicosis models, such as: tracheal intubation (tracheostomy) instillation of SiO 2 Suspension, intranasal drip and oropharyngeal inhalation of SiO 2 Suspension, using dust-staining system to inhale SiO by atomizing daily 2 Dust and other methods. [0003] Although the above methods are commonly used methods in the prior art, they all have their own advantages and disadvantages. Among them, tracheal intu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02A01K2207/30A01K2207/20A01K2227/105A01K2267/03
Inventor 李树民孙影杨方徐洪王朋李雅倩张乙
Owner NORTH CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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