CN108926708A - Allergic rhinitis mouse model clone method and application - Google Patents
Allergic rhinitis mouse model clone method and application Download PDFInfo
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Abstract
The present invention relates to a kind of allergic rhinitis mouse model clone method, include the following steps: that mouse is divided into Normal group, model control group, experimental group and positive controls first；Then suspension is injected to the mouse except Normal group, Normal group mouse is not administered；To the mouse collunarium suspension except Normal group, the mouse except Normal group is placed in closed container, Neulized inhalation is carried out with OVA normal saline solution, until there are Rhinitis Symptoms, sensitization is refused to Normal group mouse；Trial drug is given to the mouse of experimental group, dexamethasone in treatment is given to positive controls, the treatment of blank patch is given to model control group mouse, Normal group mouse is not administered；Mouse schneiderian membrance is detected again.Allergic rhinitis mouse model clone method of the invention solves the problems, such as that the duplication of mouse allelgic rhinitis models is not easy successfully to be conducive to the standardization and standardization of allergic rhinitis experimental study.
Description
Technical field
The invention belongs to preclinical medicine technical fields, and in particular to a kind of allergic rhinitis mouse model clone method and answer
With.
Background technique
The research of disease must have the patient of standard, but due to medical protection and legal requirement, in many cases
Medical research can not carry out with the patient of reality.This just needs to replicate with disease to experimental animal, establishes opposite mark
Quasi- disease animal model.Mouse has been that current medical research is more normal since small, breeding are fast and the easier feature of raising
Experimental animal has replicated successfully numerous diseases model with them.Allergic rhinitis is clinically common breathing
Tract disease, but due to being influenced by environmental factor and animal germline, the mouse model duplication of allergic rhinitis is often not easy into
Function, though success after also due to reaction and the repair mechanism of body and keep disease feature unobvious, on experimental result influence compared with
Greatly.
Summary of the invention
An object of the present invention is to provide a kind of allergic rhinitis mouse model clone methods.
Allergic rhinitis mouse model clone method of the invention, includes the following steps: S101: first dividing mouse at random
It is 4 groups, the number of every group of mouse is identical, and it is right that each group is successively designated as Normal group, model control group, experimental group and the positive
According to group；S102: and then injected daily in the mouse peritoneal of the model control group, the experimental group and the positive controls
0.15mL~0.25mL suspension is with sensitization, continuous injection 14 days；Wherein, the suspension be containing mass concentration be 8%~
The Al (OH) that 12% ovalbumin (OVA) and mass concentration is 8%~12%3Physiological saline；It is small to the Normal group
Mouse is not administered；S103: the 15 day~23 days mouse by the model control group, the experimental group and the positive controls
Collunarium is carried out with ovalbumin (OVA) and aluminium hydroxide mixed liquor, once a day, continuous 7 days；It S104: the 24-31 days will be described
The mouse of model control group, the experimental group and the positive controls is placed in transparent closed glass container, with mass concentration
Maximum atomization quantity progress Neulized inhalation is pressed for 5% OVA normal saline solution, until stopping after there are Rhinitis Symptoms；To described
Normal group mouse refuses sensitization；S105: giving trial drug to the mouse of the experimental group, and successive administration 7 days；To described
Positive controls are given dexamethasone and are injected intraperitoneally, and 1 times/day, successive administration 7 days；Blank patch is given to the model control group mouse
Agent treatment, successive administration 7 days；3.5h~4.5h is administered before Neulized inhalation；The Normal group mouse is not administered；
S106: all mouse schneiderian membrances are taken out, and are placed in the formalin that mass concentration is 10% and are fixed 22h~26h, slice direction
For skin histology block longitudal section, then HE is dyed, and is observed tissue morphology and is detected.
Allergic rhinitis mouse model clone method of the invention, disclose mouse duplication allergic rhinitis during it is thin
Section and points for attention solve the duplication of mouse allelgic rhinitis models and are not easy using standardized experiment flow and laboratory facilities
Successful problem has standardized therapeutic intervention and the timing node to make a collection of specimens after mouse allelgic rhinitis replicates successfully, has been conducive to
The standardization and standardization of allergic rhinitis experimental study.
In addition, the allergic rhinitis mouse model clone method that the present invention is above-mentioned, can also have following additional technology
Feature:
Further, in the step S101,40 mouse is chosen as subjects, are randomly divided into 4 groups, it is every group small
Mouse 10.
Further, in the step S104, taken off experimental mice the nape of the neck hair with depilatory cream (2cm~
2.5cm) × (2.5cm~3.5cm) area of size, administration range be diameter 1.8mm~2mm circular hole, each 1.8h~
2.2h。
Further, in the step S106, the detection includes to EOS quantity, the MC quantity in nasal mucosal tissue
It is measured with degranulation MC value.
Further, in the step S106, the detection further includes IgE and IL-4 content in measurement mice serum.
Further, in the step S106, the detection further includes doing disease inspection to mouse nasal mucosal tissue.
Further, in the step S104, the Rhinitis Symptoms include grabbing scratch, have a stuffy nose, sneeze, being short of breath deeply fastly
Or runny nose.
Another object of the present invention is to propose application of the method in rhinitis drug study.
Additional aspect and advantage of the invention will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Specific embodiment
The embodiment of the present invention is described below in detail, the embodiment is exemplary, it is intended to it is used to explain the present invention, and
It is not considered as limiting the invention.
Embodiment 1
Embodiment 1 proposes a kind of allergic rhinitis mouse model clone method, includes the following steps:
(1) 40 mouse are chosen first as subjects, are randomly divided into 4 groups, every group mouse 10, successively by each group mark
For Normal group, model control group, experimental group and positive controls.
(2) it is then injected daily in the mouse peritoneal of the model control group, the experimental group and the positive controls
0.15mL suspension is with sensitization, continuous injection 14 days；Wherein, the suspension be containing mass concentration be 12% OVA and matter
Measure the Al (OH) that concentration is 8%3Physiological saline；The Normal group mouse is not administered.
(3) the 15th days~23 days mouse ovum by the model control group, the experimental group and the positive controls
Albumin (OVA) and aluminium hydroxide mixed liquor carry out collunarium, once a day, continuous 7 days.
(4) the 24-31 days mouse by the model control group, the experimental group and the positive controls are placed in transparent
In closed glass container, maximum atomization quantity is pressed with the OVA normal saline solution that mass concentration is 5% and carries out Neulized inhalation, until
Appearance, which is grabbed, to be scratched, has a stuffy nose, stopping after sneeze, deep fast or runny nose symptom of being short of breath；Sensitization is refused to the Normal group mouse.
(5) experimental mice the nape of the neck hair is taken off to the area of 2.5cm × 2.5cm size with depilatory cream, administration range is
The circular hole of diameter 2mm, each 1.8h, successive administration 7 days；It gives dexamethasone to the positive controls to be injected intraperitoneally, 1 times/day,
Successive administration 7 days；It gives blank patch to the model control group mouse to treat, successive administration 7 days；3.5h gives before Neulized inhalation
Medicine；The Normal group mouse is not administered.
(6) all mouse schneiderian membrances are taken out, is placed in the formalin that mass concentration is 10% and fixes 26h, slice side
To for skin histology block longitudal section, then HE is dyed, and observation tissue morphology simultaneously does disease inspection to mouse nasal mucosal tissue, to schneiderian membrane
EOS quantity, MC quantity, degranulation MC value in tissue, IgE and IL-4 content is measured in mice serum.
Embodiment 2
Embodiment 2 proposes a kind of allergic rhinitis mouse model clone method, includes the following steps:
(1) 40 mouse are chosen first as subjects, are randomly divided into 4 groups, every group mouse 10, successively by each group mark
For Normal group, model control group, experimental group and positive controls.
(2) it is then injected daily in the mouse peritoneal of the model control group, the experimental group and the positive controls
0.25mL suspension is with sensitization, continuous injection 14 days；Wherein, the suspension be containing mass concentration be 8% OVA and quality
The Al (OH) that concentration is 12%3Physiological saline；The Normal group mouse is not administered.
(3) the 15th days~23 days mouse ovum by the model control group, the experimental group and the positive controls
Albumin (OVA) and aluminium hydroxide mixed liquor carry out collunarium, once a day, continuous 7 days.
(4) the 24-31 days mouse by the model control group, the experimental group and the positive controls are placed in transparent
In closed glass container, maximum atomization quantity is pressed with the OVA normal saline solution that mass concentration is 5% and carries out Neulized inhalation, until
Appearance, which is grabbed, to be scratched, has a stuffy nose, stopping after sneeze, deep fast or runny nose symptom of being short of breath；Sensitization is refused to the Normal group mouse.
(5) experimental mice the nape of the neck hair is taken off to the area of 2cm × 3.5cm size with depilatory cream, administration range is straight
The circular hole of diameter 1.8mm, each 2.2h, successive administration 7 days；It gives dexamethasone to the positive controls to be injected intraperitoneally, 1 times/day,
Successive administration 7 days；It gives blank patch to the model control group mouse to treat, successive administration 7 days；4.5h gives before Neulized inhalation
Medicine；The Normal group mouse is not administered.
(6) all mouse schneiderian membrances are taken out, is placed in the formalin that mass concentration is 10% and fixes 22h, slice side
To for skin histology block longitudal section, then HE is dyed, and observation tissue morphology simultaneously does disease inspection to mouse nasal mucosal tissue, to schneiderian membrane
EOS quantity, MC quantity, degranulation MC value in tissue, IgE and IL-4 content is measured in mice serum.
Embodiment 3
Embodiment 3 proposes a kind of allergic rhinitis mouse model clone method, includes the following steps:
(1) 40 mouse are chosen first as subjects, are randomly divided into 4 groups, every group mouse 10, successively by each group mark
For Normal group, model control group, experimental group and positive controls.
(2) it is then injected daily in the mouse peritoneal of the model control group, the experimental group and the positive controls
0.2mL suspension is with sensitization, continuous injection 14 days；Wherein, the suspension be containing mass concentration be 10% OVA and quality
The Al (OH) that concentration is 10%3Physiological saline；The Normal group mouse is not administered.
(3) the 15th days~23 days mouse ovum by the model control group, the experimental group and the positive controls
Albumin (OVA) and aluminium hydroxide mixed liquor carry out collunarium, once a day, continuous 7 days.
(4) the 24-31 days mouse by the model control group, the experimental group and the positive controls are placed in transparent
In closed glass container, maximum atomization quantity is pressed with the OVA normal saline solution that mass concentration is 5% and carries out Neulized inhalation, until
Appearance, which is grabbed, to be scratched, has a stuffy nose, stopping after sneeze, deep fast or runny nose symptom of being short of breath；Sensitization is refused to the Normal group mouse.
(5) experimental mice the nape of the neck hair is taken off to the area of 2.2cm × 3cm size with depilatory cream, administration range is straight
The circular hole of diameter 2mm, each 2h, successive administration 7 days；It gives dexamethasone to the positive controls to be injected intraperitoneally, 1 times/day, continuously
Administration 7 days；It gives blank patch to the model control group mouse to treat, successive administration 7 days；4h is administered before Neulized inhalation；It is right
The Normal group mouse is not administered.
(6) all mouse schneiderian membrances are taken out, is placed in the formalin that mass concentration is 10% and fixes for 24 hours, slice side
To for skin histology block longitudal section, then HE is dyed, and observation tissue morphology simultaneously does disease inspection to mouse nasal mucosal tissue, to schneiderian membrane
EOS quantity, MC quantity, degranulation MC value in tissue, IgE and IL-4 content is measured in mice serum.
Allergic rhinitis mouse model clone method of the invention, disclose mouse duplication allergic rhinitis during it is thin
Section and points for attention solve the duplication of mouse allelgic rhinitis models and are not easy using standardized experiment flow and laboratory facilities
Successful problem has standardized therapeutic intervention and the timing node to make a collection of specimens after mouse allelgic rhinitis replicates successfully, has been conducive to
The standardization and standardization of allergic rhinitis experimental study.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It can be combined in any suitable manner in one or more embodiment or examples.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the feature of different embodiments or examples described in this specification and different embodiments or examples
It closes and combines.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, modifies, replacement and variant.
Claims (8)
1. a kind of allergic rhinitis mouse model clone method, which comprises the steps of:
S101: being randomly divided into 4 groups for mouse first, and the number of every group of mouse is identical, successively by each group be designated as Normal group,
Model control group, experimental group and positive controls；
S102: and then injected daily in the mouse peritoneal of the model control group, the experimental group and the positive controls
0.15mL~0.25mL suspension is with sensitization, continuous injection 14 days；Wherein, the suspension be containing mass concentration be 8%~
The Al (OH) that 12% OVA and mass concentration is 8%~12%3Physiological saline；The Normal group mouse is not administered；
S103: the 15 day~23 days mouse egg whites by the model control group, the experimental group and the positive controls
Albumen (OVA) and aluminium hydroxide mixed liquor carry out collunarium, once a day, continuous 7 days；
S104: the 24-31 days mouse by the model control group, the experimental group and the positive controls are placed in transparent close
It closes in glass container, maximum atomization quantity is pressed with the OVA normal saline solution that mass concentration is 5% and carries out Neulized inhalation, until going out
Stop after existing Rhinitis Symptoms；Sensitization is refused to the Normal group mouse；
S105: giving trial drug to the mouse of the experimental group, and successive administration 7 days；Rice is filled in giving to the positive controls
Pine intraperitoneal injection, 1 times/day, successive administration 7 days；It gives blank patch to the model control group mouse to treat, successive administration 7 days；
3.5h~4.5h is administered before Neulized inhalation；The Normal group mouse is not administered；
S106: all mouse schneiderian membrances are taken out, and are placed in the formalin that mass concentration is 10% and are fixed 22h~26h, are sliced
Direction is skin histology block longitudal section, and then HE is dyed, and observes tissue morphology and is detected.
2. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S101
In, 40 mouse are chosen as subjects, are randomly divided into 4 groups, every group mouse 10.
3. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S105
In, experimental mice the nape of the neck hair is taken off to the area of (2cm~2.5cm) × (2.5cm~3.5cm) size with depilatory cream, is given
Medicine range is the circular hole of diameter 1.8mm~2mm, each 1.8h~2.2h.
4. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S106
In, the detection includes being measured to EOS quantity, MC quantity and the degranulation MC value in nasal mucosal tissue.
5. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S106
In, the detection further includes IgE and IL-4 content in measurement mice serum.
6. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S106
In, the detection further includes doing disease inspection to mouse nasal mucosal tissue.
7. allergic rhinitis mouse model clone method according to claim 1, which is characterized in that in the step S104
In, the Rhinitis Symptoms include grab scratch, have a stuffy nose, sneeze, deep fast or runny nose of being short of breath.
8. application of the described in any item methods of claim 1-7 in rhinitis drug study.
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Cited By (1)
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| CN111742883A (en) * | 2020-01-03 | 2020-10-09 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Construction method of lung and spleen qi deficiency allergic rhinitis combined animal model |
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