CN112889745B - Preparation method of animal model for bronchial allergic cough - Google Patents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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Abstract
The invention relates to an animal model of bronchial allergic cough induced by histamine and citric acid, which designs the animal model of the bronchial allergic cough sensitized by the histamine and the citric acid by combining the onset characteristics and the law of the bronchial allergic cough, has a larger treatment window, can play a better treatment role in both traditional antiallergic drugs and antitussive drugs, and is consistent with the clinical medication conditions of related patients. Therefore, the animal model can be used for evaluating and screening related medicines.
Description
Technical Field
The invention belongs to the field of biological medicine, and particularly relates to a method for establishing an animal model of bronchial allergic cough induced by histamine and citric acid.
Background
Bronchial allergic cough, also known as allergic bronchitis, is a clinically very common disease. The incidence of the disease is similar to that of bronchial asthma, but the disease is more affected by air pollution, and the incidence of the disease in heavily polluted areas can be more than 2 times of that of the bronchial asthma. The majority of the patients are children under 7 years old or people over 50 years old, and over 30% of children have associated dry cough and allergic cough. Often patients themselves may have a relatively clear history of allergic disease: including eczema, allergic rhinitis or allergic dermatitis, and the like. However, bronchial allergic cough has a certain similarity with bronchial allergic asthma, and specific inflammatory reactions of the trachea, bronchial mucosa and peripheral tissues can occur. Allergens, inhaled dust, atmospheric pollution, respiratory tract infections, even climate changes and catching cold can be the causative factors. However, unlike bronchial allergic asthma, bronchial allergic cough has a low probability of spasm after inflammatory reaction in the airway, and mainly involves mucus secretion and continuous constriction of the bronchial wall. The broncho-tracheal wall contracts and excess mucus blocks the bronchi, eventually causing symptoms such as allergic bronchitis and cough.
The disease is different from the traditional bronchial allergic asthma or the cough caused by irritants. It has specific immune response similar to allergic bronchial asthma, but has less occurrence of bronchospasm; it also has symptoms such as cough which allergic asthma does not have. The cough is caused by excessive mucus secretion caused by specific inflammatory reaction, and is not caused by the stimulation of chemicals to common bronchi.
Therefore, the conventional allergic asthma animal model and the cough animal model cannot completely simulate the disease state.
At present, no animal model which can completely simulate allergic cough exists in the market. Similar models are now common, including both mouse animal models of allergic bronchial asthma and models of stimulus-induced cough. In the allergic bronchial asthma model, Ovalbumin (OVA) and aluminum hydroxide adjuvant are mixed, a plurality of mice are immunized for a plurality of times, and then the mice are inhaled with atomized OVA solution for attacking. The model may ultimately trigger the symptoms associated with bronchial allergic asthma, including airway hyperreactivity due to bronchospasm, and an inflammatory response dominated by eosinophil elevation. The model of cough caused by irritants is generally prepared by directly atomizing citric acid or capsaicin and inhaling by mice to cause reflex cough. In addition, there is also a combination of the above two methods, as described in "method for establishing and testing a mouse model of CN201811207445.1 eosinophilic bronchitis", in which mice are sensitized with OVA and then challenged with methacholine nebulization to cause eosinophilic bronchitis. After superimposed citric acid stimulation, a cough response in the context of eosinophilic bronchitis occurred.
Both of the above animal models, mouse model of allergic bronchial asthma and stimulus-induced cough model, have major limitations. The mouse animal model of allergic bronchial asthma mainly changes respiratory function change caused by bronchial spasm and the like caused by activation of cholinergic receptors under the action of methacholine. The model is similar to human asthma, although the model is caused by allergen, in the process of model excitation, symptoms such as mast cell degranulation and mucus massive secretion in allergic cough do not occur generally, and cough does not occur, so the model cannot well simulate the occurrence and development process of allergic cough. The stimulus-induced cough model can only stimulate the reflex cough similar to the conditioned reflex function of human body, and does not participate in immune response. Therefore, neither model responds to the full features of allergic cough. In contrast, in the above-mentioned patent application CN201811207445.1, OVA was used for sensitization, but no secondary challenge with OVA was used and methacholine was used directly for stimulation. In this case, the airway response is still dominated by airway narrowing of asthmatic nature and allergy-related symptoms such as mucus secretion do not occur. As it is described, it causes eosinophilic bronchitis, but not bronchoallergic cough in a clinical sense.
Based on the defects of the prior art, the invention designs a model for inducing cough by citric acid after histamine sensitization by combining the onset characteristics and rules of bronchial allergic cough, the treatment window is larger, and the traditional antiallergic drugs and the antitussive drugs can play a better treatment role and are consistent with the clinical medication conditions of related patients.
Disclosure of Invention
The invention combines the onset characteristics and the law of bronchial allergic cough, designs a model for inducing cough by citric acid after histamine sensitization, and changes an animal strain from a mouse into a guinea pig which is more sensitive to anaphylactic reaction and cough. In which histamine is used to stimulate degranulation of mast cells in the airway epithelium and release inflammatory substances such as leukotriene. Under the combined action of histamine and leukotriene, goblet cells are promoted to strengthen mucus secretion such as mucin, and local pathological changes of the bronchus under the allergic condition are simulated well. And simultaneously, the citric acid is used for stimulating to induce cough. The method has wide therapeutic window for allergic cough, and the traditional antiallergic drug and antitussive drug can have good therapeutic effect, and are consistent with clinical administration conditions of relevant patients. Can be used for evaluating and screening the drug effect of related drugs.
In order to solve the technical problems, the invention provides a preparation method of an animal model of bronchial allergic cough, which is characterized in that the animal is sensitized by histamine, and the cough is induced by citric acid, so that the animal model of bronchial allergic cough is obtained.
In one embodiment, the method comprises the steps of: exposing the animal to an aerosol of citric acid and histamine to obtain the animal model of bronchial allergic cough.
In a specific embodiment, the method comprises the steps of: exposing the animal to an aerosol of 0.2-0.8M citric acid and 0.02-0.6mM histamine to obtain the animal model of bronchial allergic cough.
Preferably, the method comprises the steps of: exposing the animal to an aerosol of 0.3-0.5M citric acid and 0.06-0.2mM histamine to obtain the animal model of bronchial allergic cough.
In a specific embodiment, the method comprises the steps of: animals were exposed to an aerosol of 0.4M citric acid and 0.1mM histamine to obtain an animal model of said bronchial allergic cough.
In a specific embodiment, the exposure time is 6 to 20 minutes; preferably 8-15 minutes; more preferably 8-12 minutes and most preferably 10 minutes.
Further, the animals were exposed to an aerosol of 0.4M citric acid and 0.1mM histamine for 10 minutes at a liquid consumption rate of 0.3-0.8mL/min, preferably 0.5mL/min, via a nebulized delivery system.
On the one hand, animals used for molding need to be subjected to screening experiments: stimulating animals with citric acid and detecting cough frequency, and excluding animals with cough frequency more than 35 or less than 3; preferably, animals with a number of coughs greater than 25 or less than 15 are excluded.
Further, the animals are guinea pigs, further female Hartley guinea pigs aged 4-5 weeks.
Preferably, in one embodiment, the screening assay comprises the steps of: using a Buxco WBP system, each guinea pig was placed in a separate plethysmograph tank and exposed to 0.3-0.6M citric acid aerosol for 10 minutes at a liquid consumption rate of 0.3-0.8mL/min by a nebulization system, followed by 10 minutes of observation after the end of nebulization, the cough waveform was transmitted to MacLab/16sp via a preamplifier (MAX2, Buxco), the sound of the cough was also amplified and transmitted to MacLab/16sp, in such a way that the cough waveform and sound were recorded simultaneously on MacLab/16sp, thereby distinguishing cough from other expiratory reactions, recording the number of coughs per guinea pig, excluding animals with coughs greater than 35 or less than 3; preferably, animals with a number of coughs greater than 25 or less than 15 are excluded.
Preferably, the liquid consumption rate of the nebulized delivery system is 0.5mL/min and the citric acid aerosol concentration is 0.4M.
The invention has the beneficial effects that:
the animal model obtained by the method can effectively simulate the morbidity process, pathological manifestations and clinical symptoms of patients with bronchial allergic cough. Meanwhile, the existing clinical treatment medicine for patients with bronchial allergic asthma can embody the efficacy on the model. Solves the problem that no appropriate animal model aiming at the diseases exists in the prior art, and can provide a better animal model screening platform for the treatment drugs related to bronchial allergic cough. In particular, the present invention enables the method of the present invention to efficiently obtain a target animal model through the selection of a modeling agent and its concentration, and the selection of a method of administering the modeling agent to an animal, including exposure time, and the like.
When the mixed aerosol of citric acid and histamine is exposed, when histamine is used for stimulating the degranulation of mast cells in respiratory epithelium and releasing inflammatory substances such as leukotriene, the citric acid is used for stimulating and inducing cough, the process of inducing cough is reversed, the inflammatory substances such as leukotriene are released at an accelerated speed, goblet cells are promoted to strengthen mucus secretion such as mucin under the combined action of histamine and leukotriene, and the local pathological change of the bronchus under the allergic condition is simulated well.
In the early research process, after the high-concentration histamine is used for sensitization, under the stimulation of high histamine, mast cells in respiratory epithelium are degranulated, inflammatory substances such as leukotriene are released too fast, guinea pigs are easy to be subjected to other pathological changes, and accurate evaluation of the medicine is influenced in medicine screening as a model. The invention selects the low histamine concentration of 0.1mM to expose for 10min, so as to cause the histamine to be sequentially sensitized and reduce other concurrent lesions, thereby improving the accuracy in drug screening.
The method of the invention designs a unique animal screening experiment before modeling, and through the screening experiment, the local pathological change of the bronchus under the allergic condition of the animal model can be closer to the clinical actual pathological change condition, and the condition that the reflex cough of the conditioned reflex function induced by citric acid stimulation is too strong or too weak, which causes the pathological change condition to deviate from the clinical actual pathological change.
Drawings
FIG. 1 cough frequency in different groups of guinea pigs
FIG. 2 time to first cough in different groups of guinea pigs
Detailed Description
The technical solution of the present invention will be clearly and completely described and illustrated in the following embodiments.
The present invention will use a Buxco WBP system that includes a MAX2 preamplifier, a nebulization controller and drug delivery system, and a MacLab/16sp sensor, among others, that will be used for nebulization drug delivery and cough count analysis.
Example 1 screening of molded guinea pigs
20 female Hartley guinea pigs aged 5 weeks were selected and tested for cough sensitivity. All guinea pigs to be used were first stimulated with citric acid and tested for the number of coughs. Guinea pigs with cough numbers greater than 25 or less than 15 were excluded. The specific operation is as follows:
each guinea pig was placed in a separate plethysmograph tank and was acclimatized for 10 minutes in a fresh environment with 2L/min airflow provided by a nebulizer and flow diverter. After 10 minutes, each animal was exposed to 0.4M citric acid aerosol for 10 minutes at a liquid consumption rate of 0.5mL/min via a nebulized delivery system. The observation was continued for 10 minutes after the end of the atomization, and the total observation time was 20 minutes. Within this 20 minute period, the guinea pig cough waveform will be sent to MacLab/16sp via a preamplifier (MAX2, Buxco). The cough sound is also amplified and transmitted to MacLab/16 sp. In this way, the cough waveform and sound will be recorded simultaneously on MacLab/16 sp. This method allowed differentiation between cough and other expiratory responses, and thus recorded the number of coughs per guinea pig.
After the guinea pigs with the cough frequency not meeting the standard are removed, 16 guinea pigs meeting the conditions are screened from the guinea pigs and used for molding.
Example 2 preparation and validation of animal models
12 of the 16 guinea pigs that had been screened in example 1 and reached a closer cough were selected and after 2 weeks of recovery, the procedure was followed (4 groups of 3 guinea pigs each, with the average cough count for each group being shown before grouping in fig. 1): vehicle control (group G1) and positive drugs were given 1 hour prior to entering the plethysmographic tank: dexamethasone (group G2), methoxamine (group G3) or codeine (group G4). One hour after dosing or vehicle administration, each guinea pig was again placed in a separate plethysmograph tank, and an air stream at a flow rate of 2L/min was provided by a nebulizer and flow diverter and allowed to acclimate to the new environment for 10 minutes. After 10 minutes, each guinea pig was exposed to an aerosol of 0.4M citric acid +0.1mM histamine by a nebulizing delivery system at a liquid consumption rate of 0.5mL/min for 10 minutes. The observation was continued for 10 minutes after the end of the atomization, so the total observation time was 20 minutes. The mean number of coughs per group of guinea pigs during the period (figure 1) and the mean time to first appearance of coughs (figure 2) were recorded.
As can be seen from fig. 1, before grouping, the number of coughs was substantially comparable, with very small differences, whereas according to vehicle control (group G1) and positive drugs: after dexamethasone (group G2), methoxamine (group G3) or codeine (group G4) were grouped and exposed to 0.4M citric acid +0.1mM histamine aerosol for 10 minutes, there was no significant change in the number of coughs for the vehicle control (group G1) over the total observation period of 20 minutes, whereas administration of the positive drug showed a significant decrease.
As can be seen from fig. 2, the vehicle control (group G1) had a significantly earlier onset of first cough than the group of positive drugs.
It can be seen that upon nebulization of histamine and citric acid, the guinea pig developed a significant bronchial allergic cough. Since dexamethasone is effective in suppressing the number of coughs and prolonging the time of first cough, it is demonstrated that the model has significant allergic reactions, since the common reflex cough is unresponsive to treatment with glucocorticoids such as dexamethasone. Meanwhile, the methoxamine has better effect as a bronchus smooth muscle dilating agent with the histamine resisting effect. Codeine, as a classical central antitussive, still exerts its strongest antitussive effect. The experimental results show that the animal model of the target bronchial allergic cough is successfully obtained by adopting the molding method of the invention, and the method can be used for evaluating and screening the drug effect of related drugs.
The present invention is not limited to the embodiments disclosed in the specification and the embodiments, and the same should be considered as the disclosure of the present invention as long as it does not depart from the gist of the present invention, and equivalent embodiments that can be directly obtained by those skilled in the art from the embodiments disclosed in the specification are also within the scope of the present invention.
Claims (3)
1. A method for preparing an animal model of bronchial allergic cough comprises the following steps: exposing animals to an aerosol of 0.4M citric acid and 0.1mM histamine for 10 minutes at a liquid consumption rate of 0.5ml/min by a nebulizing delivery system, thereby obtaining an animal model of said bronchial allergic cough; animals used for molding require screening experiments: stimulating the animals with citric acid and detecting the cough frequency of the animals, and excluding the animals with the cough frequency more than 25 or less than 15; the animals are guinea pigs;
the screening experiment comprises the following steps: using the Buxco WBP system, each guinea pig was placed in a separate plethysmograph tank and exposed to 0.3-0.6M citric acid aerosol for 10 minutes at a liquid consumption rate of 0.3-0.8ml/min by a nebulisation delivery system, and observed for a further 10 minutes after nebulisation was complete, the cough waveform was sent through a MAX2 preamplifier to the MacLab/16sp sensor, and the sound of the cough was also amplified and transmitted to the MacLab/16sp sensor, in such a way that the cough waveform and sound would be recorded simultaneously on the MacLab/16sp sensor, thereby distinguishing between cough and other expiratory responses, and recording the number of coughs per guinea pig, excluding animals with coughs greater than 25 or less than 15.
2. The method for preparing an animal model of bronchial allergic cough according to claim 1, wherein the guinea pig is a female Hartley guinea pig aged 4-5 weeks.
3. The method for preparing an animal model of bronchial allergic cough according to claim 1, wherein the liquid consumption rate of the nebulizing delivery system in the screening experiment is 0.5ml/min, and the concentration of citric acid aerosol is 0.4M.
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| CN202110041741.4A CN112889745B (en) | 2021-01-13 | 2021-01-13 | Preparation method of animal model for bronchial allergic cough |
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| CN202110041741.4A CN112889745B (en) | 2021-01-13 | 2021-01-13 | Preparation method of animal model for bronchial allergic cough |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897627A (en) * | 2010-06-30 | 2010-12-01 | 广州医学院第一附属医院 | Method for establishing and detecting mouse cough model |
| CN102188442A (en) * | 2011-04-29 | 2011-09-21 | 广西中医学院 | Construction method of animal model of chronic inflammation |
| JP2018057322A (en) * | 2016-10-05 | 2018-04-12 | 国立大学法人大阪大学 | Pertussis model animal and its creation method, as well as method using pertussis model animal |
| CN109350737A (en) * | 2018-10-17 | 2019-02-19 | 广州医科大学附属第医院 | A kind of foundation and detection method of Eosinophilic bronchitis mouse model |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897627A (en) * | 2010-06-30 | 2010-12-01 | 广州医学院第一附属医院 | Method for establishing and detecting mouse cough model |
| CN102188442A (en) * | 2011-04-29 | 2011-09-21 | 广西中医学院 | Construction method of animal model of chronic inflammation |
| JP2018057322A (en) * | 2016-10-05 | 2018-04-12 | 国立大学法人大阪大学 | Pertussis model animal and its creation method, as well as method using pertussis model animal |
| CN109350737A (en) * | 2018-10-17 | 2019-02-19 | 广州医科大学附属第医院 | A kind of foundation and detection method of Eosinophilic bronchitis mouse model |
Non-Patent Citations (2)
| Title |
|---|
| 咳嗽的动物模型研究方法;陆月明;《临床肺科杂志》;20070508;第12卷(第5期);第480-481页 * |
| 诱发慢性支气管炎动物模型方法及评价;李炜等;《神经药理学报》;20190226;第9卷(第1期);第18-22页 * |
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