CN106560217A - Method for constructing allergic rhinitis model in non-human mammal by using pollen of Cryptomeria fortunei - Google Patents
Method for constructing allergic rhinitis model in non-human mammal by using pollen of Cryptomeria fortunei Download PDFInfo
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- CN106560217A CN106560217A CN201610674505.5A CN201610674505A CN106560217A CN 106560217 A CN106560217 A CN 106560217A CN 201610674505 A CN201610674505 A CN 201610674505A CN 106560217 A CN106560217 A CN 106560217A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/13—Coniferophyta (gymnosperms)
- A61K36/14—Cupressaceae (Cypress family), e.g. juniper or cypress
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- A61K39/00—Medicinal preparations containing antigens or antibodies
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- A61K39/36—Allergens from pollen
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- 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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Abstract
The invention relates to a method for constructing an allergic rhinitis model in a non-human mammal by using the pollen of Cryptomeria fortunei, belonging to the field of construction of animal models of allergic rhinitis. The pollen of Cryptomeria fortunei (Cryj1) is used as an allergen, and a solution containing Cryj1 is dripped into the nostrils of a non-human mammal so as to construct a model for allergic pollen, wherein the non-human mammal is a mouse. The allergic rhinitis model in the non-human mammal is successfully constructed in the invention, and the model realizes abnormal proliferation of epidermal goblet cells in the nasal cavities and increased secretion of mucus.
Description
Technical field
The present invention relates to allergic rhinitis animal model builds field, it is more particularly to a kind of inhuman using cryptomeria pollen structure
The method of mammal allergic rhinitis model.
Background technology
Allergic rhinitis (allergic rhinitis, AR) is that body increases to some allergen sensitivities and occurs
The allergic disease of nasal membrane, is a kind of common disease and the frequently-occurring disease in respiratory disease.According to American-European statistics, near
Allergic rhinitis incidence increased about 30% in 10 years, with the industrialized development of China, the incidence of allergic rhinitis
It is being continuously increased, is especially increasingly raising in the spring and summer pollen hypersensitivity rhinitis incidence of disease.
Cryptomeria is a kind of wide variety of plantation seeds in China and Japan, area to the east of Europe, North America and Himalaya
Nepal and the ground such as India also more plantation.In Japan, cryptomeria coverage rate up to the 18% of area, every pollen season
Section, pollinia produced by these trees sends out over long distances (send out distance can more than 100km), causes in Japan to exceed
13% population produces allergy.
Therefore, carry out being studied with regard to the Etiologic Mechanism of allergic rhinitis, inquire into various environmental factors to allergic rhinitis
Influence, and new drug spy's medicine of research and development treatment and alleviation allergic rhinitis, become extremely urgent problem demanding prompt solution.
Animal model is the important tool for studying human diseases, and setting up one kind can reflect allergic rhinitis classical symptom,
The animal model of corresponding pathological change can be reflected again, be very necessary.At present, the preparation master of allergic rhinitis animal model
If realizing by the lumbar injection of anaphylactogen, four limbs and dorsal injection and with the use of the method for adjuvant (aluminum oxide).
Said process is larger with the natural process difference of allergic rhinitis, and Neulized inhalation anaphylactogen easily causes the cause of lower respiratory tract
It is quick, asthma is easily caused after exciting.
Thus be now badly in need of it is a kind of be easy to operation there is similar pathogenesis and model configuration success rate to allergic rhinitis
High method.
The content of the invention
Present invention aim at providing the side that a kind of application cryptomeria pollen builds non-human mammal allergic rhinitis model
Method.
In order to achieve the above object, the invention provides following technical scheme：
A kind of method that application cryptomeria pollen builds non-human mammal allergic rhinitis model, with cryptomeria pollen Cryj1
As anaphylactogen, the solution containing Cryj1 is instilled from non-human mammal nostril, set up the model of pollen；Wherein, it is described
Non-human mammal is mouse.
Specially：
Step one, from first day the solution containing Cryj1 were instilled at interval of two circumferential non-human mammal nasal cavities, carried out altogether
Four times；
After step 2, above-mentioned 4th contamination, the solution containing Cryj1 was instilled to non-human mammal nasal cavity at interval of two days
Carry out altogether four times.
The nasal cavity is instilled under mouse eupnea state to nostril point nose injection, intranasal local sensitization and is excited；Often
Individual nostril respectively instills solution of the 8 μ L containing Cryj1, i.e., every mouse instills altogether 16 μ L, and the amount of liquid is not result in that Cryj1 solution is slow
Slug flow enters lung, causes the inflammatory reaction of lung.
The solution containing Cryj1 is the preparation of Jing sterile distilled waters, the μ g/ μ l of concentration 4.
The present invention builds the application of non-human mammal allergic rhinitis model can cause people to produce anaphylactoid nature
The pollen of presence sets up model.
In the method for building non-human mammal allergic rhinitis model, using Cryj1 solution mouse nostril is carried out
Point nose injection, intranasal local sensitization and excites, it is not necessary to systemic injection aluminum adjuvant etc..
In the method for building non-human mammal allergic rhinitis model, anesthetic, mouse is not used normally to exhale
Instill to nostril under suction state.
Principle is built, animal model is the important tool for studying human diseases, and the research of allergic rhinitis is also such.Reason
The preparation method of animal model is thought, on the one hand it has to be ensured that higher survival rate and life quality, method should try one's best simple easy
OK；On the other hand more require that the formation rate of pathology is high, and meet the needs of research.
Advantage for present invention：
The present invention as the method for sensitization of anaphylactogen, i.e., will contain Cryj1 molten using cryptomeria pollen Cryj1 from the instillation of mouse nostril
Liquid, and consumption volume has been efficiently controlled, inflow air flue and lung are eliminated, the possibility of respiratory tract injury is caused, meet allergy
Property rhinitis is with the unbalance pathogenesis of allergen-induced vivo immunization response.The present invention last time instill after 24 hours,
Anesthesia execution mouse, and the number of inflammatory cells from nasal lavage fluid, inflammation-associated cytokine protein expression in nose tissue, and
The aspects such as the pathological change of nose tissue are evaluated this model, and eosinophil number increases in nasal lavage fluid, nose group
Knit middle Th2 cytokines albumen to raise, non-human mammal allergic rhinitis model is successfully constructed.
The present invention makes model preferably be confined to allergic rhinitis using Cryj1 intranasals local sensitization and the method for exciting
Performance, it is to avoid described in background technology the occurrence of.The behaviouristics of the mouse model set up using the inventive method,
Biochemical Indices In Serum and histological observation meet the clinical manifestation of allergic rhinitis, and method is simple, and model survival rate is high,
Pathology rate is high.
Description of the drawings
Fig. 1 is the schematic flow sheet of the method for structure asthma model of nonhuman mammal of the present invention；Wherein, 2W：2
Week, front 4 points nose injection interval 2 weeks；2d：2 days, the 5-8 time point nose injection was spaced 2 days.
Fig. 2 is various inflammatory cell counting number result figures in nasal lavage fluid.
Fig. 3 is inflammatory cytokine protein expression of results figure in nasal lavage fluid.
Fig. 4 is mouse nasal membrane upper strata goblet cell vegetative map (PAS dyeing).
Fig. 5 is mouse nasal membrane lower floor inflammatory cell infiltration figure (HE dyeing).
Specific embodiment
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the accompanying drawing wanted needed for technology description to be briefly described.
The present invention provides a kind of method for building non-human mammal allergic rhinitis model, using Cryj1 solution, intranasal
Local sensitization and excite.
The method for building non-human mammal allergic rhinitis model provided in an embodiment of the present invention (referring to Fig. 1)：
Step one, from first day the solution containing Cryj1 were instilled at interval of two circumferential non-human mammal nasal cavities, carried out altogether
Four times；
After step 2, the 4th contamination, instilled the solution containing Cryj1 to non-human mammal nasal cavity at interval of two days and enter altogether
Row four times.
The specific implementation step of this method is：BALB/c mouse needed for experiment is weighed, control group and contamination is randomly divided into
Group, without anaesthetic treatment, direct via intranasal application instills contamination to mouse, from first day at interval of contamination in two weeks once,
Contaminating mode control group Jing or so nostril is respectively dropped into the μ L of sterile distilled water 8, contamination group Jing or so nostril and drips respectively
After entering the 4th contamination of solution of the 8 μ L containing Cryj1, contaminated once in the manner described above at interval of two days, then contaminate 4 times.
The method of described structure non-human mammal allergic rhinitis model, described non-human mammal is mouse.
The solution containing Cryj1 is cryptomeria pollen extract, is prepared using sterile distilled water, the μ g/ μ l of concentration 4.
24 hours after last time Cryj1 solution is instilled, mouse is put to death, detect inflammatory cell in its nasal lavage fluid
Number, the protein expression of inflammatory cytokine, the pathology of inflammatory cytokine protein expression and observation nose tissue in nose tissue
Learn and change (referring to Fig. 2-4 and table 1).
As seen from Figure 2 compared with control group, Cryj1 group neutrophil leucocyte numbers and eosinophil number are dramatically increased.
Fig. 3 is visible compared with control group, Cryj1 group interleukin-6s (IL-6) and horn cell chemotactic factor (CF) (KC) egg
White expression is significantly raised.
Fig. 4 and Fig. 5 are visible, and significantly, submucosa inflammatory cell infiltration is obvious for nasal membrane epithelial goblet cell hyperplasia.
Table 1 is inflammatory cytokine protein expression of results in nose tissue.Th2 inflammatory cytokines IL-5, IL-13 and acidophilus
The property granulocyte chemotaxis factor (Eotaxin) protein expression is dramatically increased, IL-6, monocyte chemoattractant protein-1 (MCP-1), KC and
The protein expression for reconciling Activated normal T cells expression and the chemotactic factor (CF) (RANTES) secreted is raised.
Inflammatory cytokine protein expression in the nose of table 1. tissue
As fully visible, mouse intranasal instills cryptomeria pollen solution, causes nasal membrane epithelial goblet cell propagation, mucus point
Secrete and increase, nasal membrane lower floor eosinophils increase, and inflammatory cell number increases and inflammatory is thin in nasal lavage fluid
Intracellular cytokine protein expression rises, and successfully constructs mouse allelgic rhinitis models.The present invention under mouse eupnea state, Jing
Nasal cavity instills micro cryptomeria pollen solution, it is to avoid it flows into the possibility for causing asthma in lung；The two weeks sensitization four in present invention interval
It is secondary, continuous agitation in 2 days four times is spaced afterwards, method is simply easy to operation；Nose tissue T h2 inflammatory cytokine IL- in the present invention
5th, IL-13 and Eotaxin protein expressions are dramatically increased, and it is to promote T lymphocytes thin to Th2 with anaphylactogen to meet allergic rhinitis
Dysuria with lower abdominal colic, the pathogenesis that inducing in vivo immune response is unbalance.
Although embodiment of the present invention is disclosed as above, it is not restricted to listed in specification and embodiment
With, it can be applied to completely various suitable the field of the invention, for those skilled in the art, can be easily
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the legend with description.
Claims (4)
1. a kind of method that application cryptomeria pollen builds non-human mammal allergic rhinitis model, it is characterised in that：With cryptomeria
Pollen Cryj1 instills the solution containing Cryj1 as anaphylactogen from non-human mammal nostril, sets up the model of pollen；Its
In, described non-human mammal is mouse.
2. the method that the application cryptomeria pollen as described in claim 1 builds non-human mammal allergic rhinitis model, it is special
Levy and be：
Step one, from first day the solution containing Cryj1 were instilled at interval of two circumferential non-human mammal nasal cavities, four are carried out altogether
It is secondary；
After step 2, above-mentioned 4th contamination, instilled the solution containing Cryj1 to non-human mammal nasal cavity at interval of two days and enter altogether
Row four times.
3. the method that the application cryptomeria pollen as described in claim 1 or 2 builds non-human mammal allergic rhinitis model, its
It is characterised by：The nasal cavity is instilled under mouse eupnea state to nostril point nose injection, intranasal local sensitization and is excited；Often
Individual nostril respectively instills solution of the 8 μ L containing Cryj1.
4. the method that the application cryptomeria pollen as described in claim 3 builds non-human mammal allergic rhinitis model, it is special
Levy and be：The solution containing Cryj1 is the preparation of Jing sterile distilled waters, the μ g/ μ l of concentration 4.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610674505.5A CN106560217A (en) | 2016-08-16 | 2016-08-16 | Method for constructing allergic rhinitis model in non-human mammal by using pollen of Cryptomeria fortunei |
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| CN201610674505.5A CN106560217A (en) | 2016-08-16 | 2016-08-16 | Method for constructing allergic rhinitis model in non-human mammal by using pollen of Cryptomeria fortunei |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101309705A (en) * | 2006-01-13 | 2008-11-19 | 独立行政法人理化学研究所 | Preventive or therapeutic agent and method for immune disease |
| CN102895657A (en) * | 2012-10-15 | 2013-01-30 | 中国人民解放军第三军医大学第二附属医院 | Method for constructing asthma model of nonhuman mammal |
-
2016
- 2016-08-16 CN CN201610674505.5A patent/CN106560217A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101309705A (en) * | 2006-01-13 | 2008-11-19 | 独立行政法人理化学研究所 | Preventive or therapeutic agent and method for immune disease |
| CN102895657A (en) * | 2012-10-15 | 2013-01-30 | 中国人民解放军第三军医大学第二附属医院 | Method for constructing asthma model of nonhuman mammal |
Non-Patent Citations (4)
| Title |
|---|
| MASAKO TSUNEMATSU等: "Establishment of an alergic rhinitis model in mice for the evaluation of nasal symptoms", 《LIFE SCIENCES》 * |
| RIE NOMIYA等: "CRTH2 plays an essential role in the pathophysiology of Cry j 1-induced pollinosis in mice", 《THE JOURNAL OF IMMUNOLOGY》 * |
| 大野泰一郎: "42例日本花粉症辨证分型和中、日两国花粉症症状及病因学比较", 《南京中医学院学报》 * |
| 殷敏等: "中国柳衫花粉症的研究近况", 《第一届全国变态反应学术研讨会论文汇编》 * |
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