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Method for constructing asthmatic animal model

An animal model and asthma technology, applied in pharmaceutical formulations, medical preparations containing active ingredients, peptide/protein components, etc., can solve problems such as large side effects and limited clinical applications

Inactive Publication Date: 2014-02-12
刘志刚 +2
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Problems solved by technology

[0004] And although adrenal glucocorticoids are still relatively effective drugs for the treatment and prevention of allergic asthma attacks, their large side effects limit the clinical application.

Method used

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Embodiment Construction

[0011] The detailed structure of the invention will be further described below in combination with specific embodiments.

[0012] 1. Preparation of silkworm protein:

[0013] Take 15-20 grams of fresh silkworm chrysalis, add liquid nitrogen to grind until it becomes powder, add 25ml of PBS silkworm chrysalis protein to wash it down, grind it in a homogenizer for 1-2 hours, take out the silkworm chrysalis protein in the homogenizer, and centrifuge at 8000rpm at 4°C / 15min, take the supernatant, repeat centrifugation three times, and measure the protein concentration by BCA method to reach the standard, and obtain the silkworm protein.

[0014] 2. Modeling:

[0015] Babl / c mice aged 5-6 weeks were sensitized by intraperitoneal injection of silkworm chrysalis protein 50 μg / mouse and aluminum hydroxide 4 mg, once a week, and continuous intraperitoneal injection for three weeks. After the last sensitization, the challenge was continued for one week (50 μg of silkworm chrysalis cr...

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Abstract

The invention provides a method for constructing an asthmatic animal model. The method comprises the following steps: grinding 15-20g of fresh silkworm chrysalis into powder with liquid nitrogen; adding 25ml of PBS (phosphate buffer solution) to wash silkworm protein, and grinding the silkworm protein for 1-2 hours in a homogenizer; taking out the silkworm protein in the homogenizer, and centrifuging at 4 DEG C at 8,000rpm for 15 minutes; repeatedly centrifuging supernatant for three times; assaying whether protein concentration reaches standard or not by adopting a BCA (bicinchoninic acid) method to obtain the silkworm protein. The silkworm protein is applied to a Babl / c mouse at 5-6 weeks, injecting 50mu g of silkworm protein into a mouse in an intraperitoneal manner, and adding 4mg of aluminum hydroxide for sensitization once a week, and continuously performing intraperitoneal injection for three weeks; continuously exciting for 1 week (50mu g of silkworm crude protein is inhaled in a nasal manner for a mouse) after the last time of sensitization; performing airway hyperreactivity assay and pathological examination in 24 hours after nasal inhalation in the last day. According to the method, the problem of asthmatic animal model construction difficulty can be solved, and the animal model is stable and controllable and can be used for studying asthma pathogenesis and curative effect evaluation of asthma treatment medicaments.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically, the invention relates to a method for constructing an animal model of asthma. Background technique [0002] Allergic asthma seriously affects human health, and its incidence is increasing at a rate of 5% every year. According to the estimates of the United Nations World Health Organization, there are as many as 300 million asthma patients worldwide, and it affects people of all levels including different ages or different economic conditions. Allergic diseases like asthma have been on a steady rise over the past two decades. This not only affects people's quality of life, especially children's asthma, which directly affects the normal development of children's bodies, but also hinders the development of the country's economy. [0003] At present, research on allergic asthma is deepening at home and abroad, but there are relatively few effective and specific treatments for all...

Claims

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Application Information

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IPC IPC(8): A61K38/02
Inventor 刘志刚左建宏李维中
Owner 刘志刚
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