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Trichosanthin protein induced mouse allelgic asthma disease model

A technology of trichosanthin and allergic asthma, applied in the biological field, can solve unclear problems

Inactive Publication Date: 2009-08-05
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although the three-signal theory that DC induces Th1 or Th2 responses has been basically recognized, that is, MHCII-TCR pathway; co-stimulatory factor pathway; Which key molecules play a role is not very clear in this field, and its specific mechanism still needs more systematic and in-depth research

Method used

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  • Trichosanthin protein induced mouse allelgic asthma disease model
  • Trichosanthin protein induced mouse allelgic asthma disease model
  • Trichosanthin protein induced mouse allelgic asthma disease model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1 Establishment of Mouse Allergic Asthma Model

[0096] 1. Establish mouse allergic asthma model induced by trichosanthin and aluminum adjuvant (model 1)

[0097] C57BL / 6 mice (female, 6-8 weeks), were intraperitoneally injected with PBS solution of trichosanthin (TCS, 99% pure Trichosanthes extract, purchased from Shanghai Jinshan Pharmaceutical Co., Ltd.) on the 1st day and 14th day, the dose was 5ug / 200ul PBS / mouse, mixed with 1 mg aluminum adjuvant (Alum, suspension of aluminum hydroxide in PBS, containing 10% aluminum hydroxide). From the 21st day to the 23rd day, the mice were challenged with the respiratory tract of the antigen for 3 consecutive days, 30 minutes a day, and the PBS solution containing TCS was nebulized with an ultrasonic nebulizer at a dose of 2 mg / ml. Twenty-four hours after the last antigen challenge, mice were dissected for testing. Using the same treatment method, mice were treated with the same volume of PBS solution containing alum...

Embodiment 2

[0104] Example 2 Mouse allergic asthma model test

[0105] Each mouse model prepared in Example 1 was tested, and the tested indicators included: the total number of inflammatory cells infiltrating into the airway, the number of eosinophils, the level of IgE in the serum and the pathological section of lung tissue. The amount of total inflammatory cells and eosinophils infiltrated in pulmonary bronchial lavage fluid can verify whether TCS causes allergic airway inflammation.

[0106] The detection method is as follows: blood is taken from the mouse eyeball, blood is collected to collect serum; the mouse is killed, the left lung is ligated, the trachea is separated, the trachea is inserted into the mouse trachea with a No. 9 needle, and 0.5ml of PBS solution is injected in 3 times to lavage the airway and suck out , the number of infiltrating cells and cytokines to be measured in the lavage fluid. The left lung was separated for pathological sectioning. The amount of IgE in s...

Embodiment 3

[0112] Example 3 Trichosanthin causes Th2 type immune response in vivo

[0113] In this example, the type of allergic asthma induced by TCS was determined by detecting the content of Th2 type cytokines IL-4, IL-5 and IL-13 in the lung lavage fluid of the animal model induced by trichosanthin. The PBS control group was used as the control.

[0114] Detection of IL-4, IL-5, IL-13, INF-γ, IgE, IgG1, IgG2a was performed according to conventional methods.

[0115] see results image 3 , wherein, the expression of cytokines IL-4 (A), IL-5 (B), IL-13 (C), and INF-γ (D) was measured in bronchial lavage fluid (BALF); lung draining lymph node cells were tested in vitro The expression of cytokines IL-4 (E), INF-γ (F) was measured after re-stimulation with TCS (LNs). The levels of immunoglobulins in serum were measured as total IgE (G) and TCS-specific IgG2a (H). It can be seen that in the bronchial lavage fluid (BALF) of the animal model induced by trichosanthin (with or without adju...

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Abstract

The invention belongs to the field of biotechnology and discloses application of trichosanthin in preparing an allelgic asthma model of mice, wherein the trichosanthin can exert functions without the assistance of an adjuvant. The invention also discloses an allelgic asthma disease model of the mice which is induced by the trichosanthin. An allelgic asthma animal model established by the invention has typical characteristics of an allelgic asthma disease, and can well simulate the generation and development of allelgic asthma in vivo. At the same time, the animal model can also be applied to screening medicines against the allelgic asthma.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically, the invention relates to the use of trichosanthin in preparing a mouse allergic asthma model, and a mouse allergic asthma disease model induced by the trichosanthin. Background technique [0002] 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. Although the research on allergic asthma at home and abroad is deepening day by day, there are relatively few effective and specific treatments for allergic asthma. Now the internationally accepted animal model of allergic asthma is only the mouse asthma model induced by chicken ovalbumin (OVA). Since OVA is not an allergen that causes diseases in humans, the research results obtained through this mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00G01N33/48
Inventor 孙兵王媛毛开睿
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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