CN109381278A - A kind of method for building up of cerebral damage rat animal model - Google Patents
A kind of method for building up of cerebral damage rat animal model Download PDFInfo
- Publication number
- CN109381278A CN109381278A CN201710667200.6A CN201710667200A CN109381278A CN 109381278 A CN109381278 A CN 109381278A CN 201710667200 A CN201710667200 A CN 201710667200A CN 109381278 A CN109381278 A CN 109381278A
- Authority
- CN
- China
- Prior art keywords
- rat
- animal model
- cerebral damage
- method described
- group
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D7/00—Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to radiation protection technique fields, are related to a kind of method for building up of cerebral damage rat animal model.The method is using the head local irradiation of ray single, and then to form cerebral damage animal model for after rat anesthesia.Utilize the method for building up of cerebral damage rat animal model of the invention, it is established that the rat model come can preferably simulate human body by radiation-induced cerebral damage.
Description
Technical field
The invention belongs to radiation protection technique fields, are related to a kind of foundation side of cerebral damage rat animal model
Method.
Background technique
Currently, radiotherapy is the important method for treating incidence primary tumo(u)r and metastatic tumour.Due to three-dimensional conformal, adjust
The maturation and popularization of the modernization radiation technique such as strong conformal and γ (X) knife, the survival rate of head and neck neoplasm radiotherapy group are improved, are deposited
Live time extends.But radiotherapy also can inevitably damage in range of exposures normal group while injuring tumor cell
It knits, so as to cause radioactive damage, typically life cycle is more than 6 months patients, and have 90% cognition dysfunction can occur,
Wherein the long-term survivors of 2%-5% can develop in turn as dementia.
Cognition dysfunction can behave as study, memory, thinking, information processing, reasoning, judgement, language expression etc.
Aspect is abnormal, seriously affects patients ' life quality.
There are many method for building up of cerebral damage model, such as rat disclosed in CN101107924A injects half Guang of homotype
Sour method, mouse infection human cytomegalovirus method disclosed in CN1724075A, lateral ventricle of rat brain/hippocampus disclosed in CN1636601A
Inject protein kinase agonist/inhibitor method.But above-mentioned modeling method cannot simulate radiation-induced cerebral damage.
Summary of the invention
The object of the present invention is to provide a kind of method for building up of cerebral damage rat animal model, big can establish
Mouse model preferably simulates human body by radiation-induced cerebral damage.
In order to achieve this, the present invention provides a kind of cerebral damage rat animal mould in the embodiment on basis
The method for building up of type, the method using the head local irradiation of ray single-bolus high-dose, and then are formed after rat anesthesia
Cerebral damage animal model.
The verification method for the cerebral damage rat animal model that the present invention establishes is: using Morris water maze laboratory
(orientation navigation experiment+space search experiment) carries out the detection of learning and memory in rats ability, is dyed using HE and carries out cerebral morphology
Learn observation.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the rat is SD rat.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the rat is 4-5 week old male underage rat.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the anesthesia is to use concentration for the chloraldurate of 100g/L, every rat passes through according to 0.3ml/100g dosage
Intraperitoneal injection is anaesthetized.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the ray is 4-8MeV electric wire.
In a kind of more preferred embodiment, the present invention provides a kind of building for cerebral damage rat animal model
Cube method, wherein the electric wire dosage rate is 2-4Gy/min, exposure dose 10-30Gy.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the head is locally rat eye outer canthus to ear rear range.
In a preferred embodiment, the present invention provides a kind of foundation side of cerebral damage rat animal model
Method, wherein the cerebral damage Animal Model time is 4-6 weeks.
The beneficial effects of the present invention are, using the method for building up of cerebral damage rat animal model of the invention,
The rat model set up can preferably simulate human body by radiation-induced cerebral damage.
The model constructed in the process of the present invention can represent the real process that radiation causes cerebral damage, the formation of model
Process meets the actual conditions of clinical patients, and model built can be used for screening new drug, evaluation clinical therapeutic efficacy and to study
Pathogenesis etc..In addition, the present invention also has the features such as easy to operate, favorable reproducibility, required cost is low.
Detailed description of the invention
Fig. 1 for the cerebral damage rat animal model established the rat in Morris water maze test escape it is latent
Phase trend chart.
The rats'swimming track in Morris water maze test for the cerebral damage rat animal model established Fig. 2
Schematic diagram, wherein four figures indicate different exposure dose of radiation group rat learning periods swimming track schematic diagrams above, below four
Figure indicates different exposure dose of radiation group Rats With Memory phases swimming track schematic diagrams.
Fig. 3 is the pathological section testing result for the cerebral damage rat animal model established.In figure: A is control
Group, B are 10Gy IR group, and C is 20Gy IR group, and D is 30Gy IR group；Solid arrow meaning is apoptotic body, and dotted arrow is signified
For the area Kong Ran.
Specific embodiment
A specific embodiment of the invention is further illustrated with reference to embodiments.
Embodiment 1: animal radiation irradiation
SD rat, SPF grades, male, 4-5 week old, by Test Animal Centre, Academy of Military Medical Sciences, P.L.A
It provides, production licence number: SCXK- (army) 2012-0004, quality certification number: 0035516.
Animal is randomly divided into the simple irradiation group (10Gy of control group (Control, C), 10Gy according to weight after quarantine in 6 days
IR), the simple irradiation group of 20Gy (20Gy IR), the simple irradiation group of 30Gy (30Gy IR), every group 5.
(1) control group: every animal does not receive radiation irradiation, gives purified water, volume 20ml/kg through stomach-filling；
(2) the simple irradiation group of 10Gy: every animal receives the irradiation of 10Gy single irradiation；
(3) the simple irradiation group of 20Gy: every animal receives the irradiation of 20Gy single irradiation；
(4) the simple irradiation group of 30Gy: every animal receives the irradiation of 30Gy single irradiation.
Specific radiation illuminating method is as follows.
Rat body weight is weighed, every rat passes through intraperitoneal injection of anesthesia agent (concentration 100g/ according to 0.3ml/100g dosage
The chloraldurate of L) it is anaesthetized.The rat anaesthetized is fixed on cystosepiment, with (the Shandong Xinhua medical treatment of X-ray analog machine
Instrument limited liability company, SL-IE type) positioned, irradiated site is head eyes outer canthus to ear rear range, body its
He is partially covered with stereotype.Then give dosage rate be 3Gy/min 6MeV electron beam irradiation, respectively according to 10Gy, 20Gy,
The exposure dose of 30Gy carries out single irradiation irradiation.
Embodiment 2: the detection test of animal after radiation irradiation
1, test apparatus
Morris water maze (institute of Materia Medica,Chinese Academy of Medical Sciences), medical computerized linear accelerator synergy
(Elekta Limited), paraffin wax embedding Histocentre 3 (Shandon), paraffin slicing machine Finesse 325
(Shandon), Automation-tissue-dehydrating machine Excelsior ES (Shandon), fluorescence microimaging systems BX53
(Olympus)。
2, Morris water maze test
I) test method
Rat radiation irradiation the latter moon carries out Morris water maze test.Morris water maze is cylindrical by black inner wall
Pond and a position-movable and height-adjustable plastics platform composition.Diameter 100cm, high 50cm, pool inner water depth 30cm,
Heater heats water in (21 ± 2) DEG C or so.Pond indicates the place of entry of east, south, west, north four, and pond is divided into northeast
(NE), the southeast (SE), northwest (NW), southwestern (SW) four quadrants are flat away from an escape is put at pool side edge 20cm in SE quadrant center
Platform, platform high 29cm, diameter 10cm, this platform surface have protrusion to stand convenient for animal, and Chi Zhongshui did not had platform 1cm or so,
Pool wall has various patterns to provide space referring to clue for animal in pond, is convenient for rat locating platform, places and take the photograph above water maze
Camera, the interior installation software of computer tracks and records rats'swimming track, for statistical analysis to data after the completion.
2.1 orientation navigation experiment
Orientation navigation experiment starts free swimming in the pond that rat is put into non-placement platform by the previous day in formal experiment
2min is familiar with experimental situation.The orientation navigation experiment period is 5 days, daily 1 period of training, each period training 4 times, respectively
Enter water from 4 place of entry of 4 quadrants, platform is placed in SE quadrant before training, rat is put into towards pool wall from place of entry
Chi Zhong tracks and records the swimming track of rat, and records it from water is entered to climbing to platform (four limbs are on platform)
Time, this time are escape latency.Rat is allowed to put back in cage after the rest several seconds on platform.If rat in 2min not
Platform is found, then platform is led to by experimenter, escape latency is denoted as 120s.4 escapes are latent in each period
Achievement of the mean as this period that count of volt phase, and counted.
The experiment of 2.2 space search
Space search experiment carries out after orientation navigation experiment, i.e., removes within the 6th day platform, and an optional place of entry is by rat
It is put into pond, records the swimming track of rat in 1min and is analyzed.Its time for passing through original platform position of observation analysis
Number, record rat is in original platform quadrant residence time, platform quadrant swim distance and spanning platform number, to detect the sky of rat
Between memory capability.
II) test result
All data are analyzed using SPSS16.0 statistical analysis software below, and total data is with mean ± standard deviation
(means ± SD) is indicated.
Morris water maze laboratory 5 days orientation navigation experiment result such as Fig. 1 and table 1.From figure 1 it appears that the time
Factor (day) is statistically significant (P < 0.05), illustrates that escape latency has the tendency that changing over time；But time and grouping
Reciprocation is not statistically significant (P > 0.05), illustrates that the effect of time factor is not different with the difference of grouping.Individual
Between make a variation part calculated result show that the P value of grouping illustrates that grouping factor works less than 0.05, each group incubation period index
It is different in general.Shown in table 1,30Gy IR group in 1-5 days training periods, latency be above its excess-three group and
Difference has statistical significance (P < 0.05).
The different exposure dose group treated rats in Morris water maze performance directed navigation experimental results of table 1
Note: P < 0.05 compared with the control group *.
Space search experimental result is as shown in table 2 within 6th day, and 30Gy IR group is compared with remaining each group original platform quadrant residence time
It is significant to shorten (P < 0.05)；Spanning platform number substantially reduces (P < 0.05)；10Gy IR group, 20Gy IR group, 30Gy IR group are former
Platform quadrant swim distance significantly shortens (P < 0.05) compared with C group, and with the increase of dosage, original platform quadrant swim distance by
It is tapered short.Each group rats'swimming track record is shown in Fig. 2.Lines are rats'swimming track in Fig. 2, and circle is platform position,
Rat climbs to track record end on platform, can be according to the more different intervention group escape latencies of track record, original platform quadrant
Residence time, platform quadrant swim record and cross over platform number.
The different exposure dose group treated rats in Morris water maze performance space exploration experimental results of table 2
Note: P < 0.05 compared with the control group *.
3, pathological section detects
I) pathological section production and HE colouring method
(1) film-making: repairing and take brain, routinely dehydration, transparent, waxdip, embedding, slice and HE dyeing.
(2) it is dehydrated and embeds: taking tissue, fixer is rinsed through tap water, (70%, 80%, 95% after alcohol serial dehydration
I, 95% II, 100% I, 100% II), carries out transparent (dimethylbenzene I, dimethylbenzene II) to tissue, waxdip (paraffin I, paraffin II)
After embed.
(3) it is sliced: 3~5 μm of tissue slices after embedding, through 60 DEG C of roasting piece poststainings.
(4) dye: HE dyeing (dimethylbenzene I, dimethylbenzene II dewaxing, cross 100% I, 100% II, 95% I, 95% II,
80%, 70% graded ethanol, haematoxylin dyeing, differentiation, anti-indigo plant, eosin stains, mistake 70%, 80%, 95% I, 95% II,
100% I, 100% II graded ethanol).
(5) transparent and mounting: gummy mounting after dimethylbenzene I, dimethylbenzene II are transparent.
II) testing result
The pathological change of various dose group rat brain Hippocampus CA 3 Region
Testing result is shown in Fig. 3.HE dyeing optics microscopically observation is shown:
(1) C group Hippocampus CA 3 Region nerve cell does not observe the pathological changes such as apoptosis, necrosis substantially, and cellular morphology is normal,
Cell is completely embedded, and marshalling.
(2) most of centrum confluent monolayer cells endochylema shrinkage, deep dye, cell quantity are reduced after irradiating, and level disorder arranges nothing
Sequence, sparse, loose, cell connection is not close.
(3) the visible apoptotic body of the individual centrum layer inner cells of 10Gy IR group, a small amount of neuronal cell death, structure disappear；
20Gy IR group and the destruction of 30Gy IR group institutional framework are more serious compared with 10Gy IR group, and it is aobvious histocyte apoptosis, necrosis etc. occur
The pathological change of work；And there is the area Kong Ran in 30Gy IR group, it is seen that multiple apoptotic bodies.
4, conclusion
It is irradiated by 6MeV electric wire 10-30Gy single irradiation, successfully constructs the rat animal mould of cerebral damage
Type.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
Mind and range.If in this way, belonging to the model of the claims in the present invention and its equivalent technology to these modifications and changes of the present invention
Within enclosing, then the present invention is also intended to include these modifications and variations.Above-described embodiment or embodiment are only to the present invention
For example, the present invention can also be implemented with other ad hoc fashions or other particular form, without departing from of the invention
Main idea or substantive characteristics.Therefore, the embodiment of description is regarded as illustrative and non-limiting in any way.This
The range of invention should be illustrated that any variation equivalent with the intention and range of claim also should include by appended claims
Within the scope of the invention.
Claims (8)
1. a kind of method for building up of cerebral damage rat animal model, it is characterised in that: the method is by rat fiber crops
After liquor-saturated, using the head local irradiation of ray single, and then cerebral damage animal model is formed.
2. according to the method described in claim 1, it is characterized by: the rat is SD rat.
3. according to the method described in claim 1, it is characterized by: the rat is 4-5 week old male underage rat.
4. according to the method described in claim 1, it is characterized by: the anesthesia is the hydration chlorine for using concentration as 100g/L
Aldehyde, every rat are anaesthetized according to 0.3ml/100g dosage by intraperitoneal injection.
5. according to the method described in claim 1, it is characterized by: the ray is 4-8MeV electric wire.
6. according to the method described in claim 5, it is characterized by: the electric wire dosage rate is 2-4Gy/min, irradiation agent
Amount is 10-30Gy.
7. according to the method described in claim 1, it is characterized by: after the head is locally rat eye outer canthus to ear
Edge range.
8. according to the method described in claim 1, it is characterized by: the cerebral damage Animal Model time be
4-6 weeks.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710667200.6A CN109381278A (en) | 2017-08-07 | 2017-08-07 | A kind of method for building up of cerebral damage rat animal model |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710667200.6A CN109381278A (en) | 2017-08-07 | 2017-08-07 | A kind of method for building up of cerebral damage rat animal model |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109381278A true CN109381278A (en) | 2019-02-26 |
Family
ID=65413506
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710667200.6A Pending CN109381278A (en) | 2017-08-07 | 2017-08-07 | A kind of method for building up of cerebral damage rat animal model |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109381278A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110367190A (en) * | 2019-07-18 | 2019-10-25 | 中国辐射防护研究院 | A kind of method of Liver Damage in Rats model foundation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203208555U (en) * | 2013-04-28 | 2013-09-25 | 复旦大学附属肿瘤医院 | Irradiation positioning device for animal experiment |
-
2017
- 2017-08-07 CN CN201710667200.6A patent/CN109381278A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203208555U (en) * | 2013-04-28 | 2013-09-25 | 复旦大学附属肿瘤医院 | Irradiation positioning device for animal experiment |
Non-Patent Citations (2)
| Title |
|---|
| 孙锐: "低剂量全脑照射引起不同年龄大鼠认知功能损害的初步实验研究", 《中国优秀硕士学位论文全文数据库（电子期刊）医药卫生科技辑》 * |
| 韩润润: ""放射性脑损伤对认知功能的影响及机制初探"", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110367190A (en) * | 2019-07-18 | 2019-10-25 | 中国辐射防护研究院 | A kind of method of Liver Damage in Rats model foundation |
| CN110367190B (en) * | 2019-07-18 | 2022-09-27 | 中国辐射防护研究院 | Method for establishing rat liver injury model |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107913467A (en) | For will travel across the visual apparatus and method in hadron beam path of destination organization by magnetic resonance imaging | |
| Sun et al. | Experience-dependent structural plasticity at pre-and postsynaptic sites of layer 2/3 cells in developing visual cortex | |
| Hescham et al. | Fornix deep brain stimulation induced long-term spatial memory independent of hippocampal neurogenesis | |
| EP3547830A1 (en) | Method and device for combating salmon lice and other ectoparasites in fish | |
| Ali et al. | Environmental enrichment enhances systems-level consolidation of a spatial memory after lesions of the ventral midline thalamus | |
| Shukitt-Hale et al. | The effects of proton exposure on neurochemistry and behavior | |
| Amaroli et al. | Paramecium: a promising non-animal bioassay to study the effect of 808 nm infrared diode laser photobiomodulation | |
| CN109523586A (en) | A kind of dosage rate evaluation method and system based on Monte Carlo voxel model | |
| CN107913468A (en) | Include the Medical Devices of hadron therapy device, MRI and hadron X-ray camera system | |
| Zhao et al. | Changes in neurons and synapses in hippocampus of streptozotocin‐induced type 1 diabetes rats: a stereological investigation | |
| CN109381278A (en) | A kind of method for building up of cerebral damage rat animal model | |
| Jahanshahi et al. | Radiation-induced lowered neurogenesis associated with shortened latency of inhibitory avoidance memory response | |
| Katz et al. | Whole-organism 3D quantitative characterization of zebrafish melanin by silver deposition micro-CT | |
| Yoon et al. | A precision 3D conformal treatment technique in rats: Application to whole‐brain radiotherapy with hippocampal avoidance | |
| Lau et al. | Effects of visual or light deprivation on the morphology, and the elimination of the transient features during development, of type I retinal ganglion cells in hamsters | |
| DE112013004939T5 (en) | Tumor therapy by targeted nuclear fusion | |
| Sasaki et al. | A simulation study of the radiation-induced bystander effect: modeling with stochastically defined signal reemission | |
| Fedotov et al. | Quantitative cognitive‐test characterization of reconnectable implantable fiber‐optic neurointerfaces for optogenetic neurostimulation | |
| Semmler et al. | An efficient method for fractionated whole rodent brain radiation | |
| Koziak et al. | Light alters nociceptive effects of magnetic field shielding | |
| CN110367190B (en) | Method for establishing rat liver injury model | |
| Dos Santos et al. | Development of whole brain versus targeted dentate gyrus irradiation model to explain low to moderate doses of exposure effects in mice | |
| DE102017000809B3 (en) | Apparatus and method for controlling salmonidae and other ectoparasites in fish | |
| Hu et al. | The morphological changes of pyramidal and spiny stellate cells in the primary visual cortex of chronic morphine treated cats | |
| CN108354961B (en) | Application of epimedium extract in preparing medicine for treating postradiation cranial nerve injury and cognitive disorder |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |