An in vitro animal trachea model establishment method and evaluation method for evaluating tobacco mainstream smoke-induced inflammatory response

Abstract

The invention relates to a method for establishing an animal in-vitro tracheal model for evaluating inflammatory response induced by tobacco mainstream smoke and an evaluation method, and belongs to the technical field of cigarette hazard assessment. The method for establishing the animal in-vitro tracheal model comprises the following steps: 1) separating a tracheal ring; 2) culturing the tracheal ring obtained in the step 1) in vitro; and 3) exposing the tracheal ring cultured in the step 2) to a flue gas condensate for 1 to 24 hours. The evaluation method of the inflammatory response induced by the tobacco mainstream smoke comprises selection of evaluation indexes and division of evaluation criteria; and evaluation of the degree of the inflammatory response. The method has the followingadvantages: 1) the detection method of the tracheal inflammation after smoke exposure has high efficiency; 2) the detection method of the tracheal inflammation after smoke exposure has good repeatability and system stability and high accuracy; and 3) the use range is wide. The method can be applied to in vitro exposure models of cigarette mainstream smoke, sidestream smoke, ambient flue gas and other smoke aerosols.

Description

technical field [0001] The invention relates to an animal in vitro trachea model establishment method and an evaluation method for evaluating tobacco mainstream smoke-induced inflammation, and belongs to the technical field of cigarette hazard evaluation. Background technique [0002] Smoking can cause inflammation of the trachea and lungs and is the primary risk factor for chronic obstructive pulmonary emphysema (COPD). Through the in vitro experiments of the trachea, the mechanism and process of the inflammatory response can be preliminarily understood. Cigarettesmoke (CS) causes tissue damage in the airways, which has been implicated in the development of CS-induced airway disease and related lung diseases, such as chronic obstructive pulmonary disease (COPD) (Murray LA ,Dunmore R,Camelo A,Da Silva CA,Gustavsson MJ,Habiel DM,Hackett TL,Hogaboam CM,Sleeman MA,Knight DA.Acute cigarette smoke exposure activates apoptotic and inflammatory programs but a second stimulus is re...

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Problems solved by technology

[0003] However, due to the long cycle of establishing COPD models in animal experiments and many influencing factors, it is difficult to compare the results of various studies precisely because of the lack of standardized smoke exposure and standardized analysis, including different types of cigarettes, cigarette doses, Various influencing factors such as research equipment, exposure design and a wide range of mouse strains (smoke plus tracheal instillation of bleomycin to prepare a rat model of chronic obstructive pulmonary disease and the intervention effect and mechanism of resveratrol on the disease Zhou Min's doctoral dissertation 2008), so it is very necessary to use in vitro models to further study the mechanism of airway inflammation after CS exposure

[0004] At present, most research methods to solve the rapid COPD risk model assessment in vitro are two-dimensional cultured cell models, but because the two-dimensional model limits intercellular reactions, destroys cell organization and polarity, cannot accurately represent the growth environment of cells in vivo, and disconnects cells in vivo. The relationship between in vitro performance and other defects has caused a certain impact on research (Haycock, J.W. 3D CellCulture: A Review of Current Approaches and Techniques. Methods Mol. Biol. 2011, 695, 1– 15.)

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