CN1660446A - Method and application of producing disease model of local cerebral ischemia by using macaque - Google Patents
Method and application of producing disease model of local cerebral ischemia by using macaque Download PDFInfo
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Abstract
A focal cerebroischemia model for researching and treating human cerebral thrombus is created through providing a macaca mulatta, and introdicing a microspherical catheter to block the start position of cerebral artery.
Description
Technical field
The invention belongs to medical science and life sciences experimental study field, be specifically related to a kind of method of making primate focal cerebral ischemia disease model.
The design has overcome traditional primate cerebral ischemia method Ischemia Time and the out of contior shortcoming of degree, reasonable in design, novel, be convenient to popularization of hospital and scientific research department and use, provide necessary primate model for the mankind defeat cerebral thrombosis, thereby carry out the research of pathogenetic discussion and therapeutical effect.
Background technology
The Mechanism Study of cerebral ischemia has had the history of last 100 years, and remains one of present neuroscience important research project.Because the restriction of experiment condition, the zoopery object of former studies cerebral ischemia generally is confined to animals such as rat, rabbit, and primate is less relatively, but animals such as rat are huge with human difference after all.In contrast to this, the macaque and the mankind belong to primates together, and its biological characteristics is with human close, and is even more ideal as the experimental subject of research cerebral ischemia.
At present the most frequently used animal model of research focal cerebral ischemia is middle cerebral artery occlusion/irritate an again model, need be outside blood vessel or the blood flow of intravascular occlusion middle cerebral artery.The distribution of macaque middle cerebral artery only has small difference with the distribution of the mankind's middle cerebral artery in the dissection, and just the body of human brain is bigger; And the pathological change that the monkey middle cerebral artery occlusion is irritated again is very similar to the variation of people's apoplexy.
Documents and materials show that the macaque middle cerebral artery occlusion irritates model again and mainly contain through eye socket and open the cranium method, directly open cranium middle cerebral artery occlusion method.It is many more directly to have opened the progress of cranium method through the eye socket method, can reduce cerebrospinal leak relatively.This method proposed as far back as 1970, and the reliability of this model is by many laboratorys confirmations in the world.But some significant disadvantages of the existence of this method:
1. need operation in the meticulous eye socket.
2. damaged the nerve net on the middle cerebral artery.
3. because cranial cavity is open, cause cerebrospinal leak.
4. be in the operation of dark chamber during owing to operation, in order to close middle cerebral artery with apparatuses such as vascular clamp folder, when separating middle cerebral artery, circumvascular arachnoidea is impaired, so causes oozing of blood easily, thereby causes vasospasm, influences the logical again of blood vessel.
5. the blood flow that can not guarantee blood vessel is stable, is unable to estimate the situation of collateral circulation, also is difficult to control the size of focus of infarct.
Therefore, the someone thinks that even perform the operation suitable standardization, the perioperatively physiological condition is controlled fairly goodly, and the degree of cerebral ischemia also has very big difference.The variation of collateral circulation may be to produce the inconstant reason of ischemic focus.Therefore, the variation of brain local circulation on time and space makes that focus of infarct is difficult to use statistical procedures, is difficult to be used for assessing therapeutic effect such as medicine.
Summary of the invention
The objective of the invention is to overcome above-mentioned the deficiencies in the prior art, is object with the experiment macaque, and by special microsphere ductus bursae, the innovation blocking method has been studied a kind of method of making the focal cerebral ischemia disease model with macaque.A kind of new macaque middle cerebral artery occlusion/irritate again model is proposed, the new approaches that the middle cerebral artery of macaque is carried out the obstruction of controlled time-histories and irritated.
Technical scheme of the present invention is:
Research and propose and a kind ofly make the method for the disease model of focal cerebral ischemia, it is characterized in that with macaque:
Test animal and choose primate macaque Macaca mulatta;
Get involved the microsphere ductus bursae in the blood vessel, the initial part of blocking middle cerebral artery makes middle cerebral artery blood supply district ischemia by requirement of experiment, makes the cerebral ischemia disease model.
Getting involved microsphere ductus bursae method in the above-mentioned blood vessel is:
(1) operation gets involved sacculus obstruction laboratory animal middle cerebral artery blood supply district
Insert the special microsphere ductus bursae that soaked with heparin in advance from the macaque external carotid artery, insert middle cerebral artery through internal carotid artery, be inflated to the full initial part of blocking middle cerebral artery of microsphere capsule to the microsphere ductus bursae, make middle cerebral artery blood supply district ischemia continue 2h, make the cerebral ischemia disease model;
(2) ischemic region is irritated again
By the interval of requirement of experiment, emit gas from the microsphere ductus bursae, extract the microsphere ductus bursae under the local anaesthesia out, ischemic region is irritated again;
Operation finishes, and after stitching was handled, laboratory animal was positioned in 25 ℃ of air-conditioned rooms and rests;
(3) record, scoring
Carry out neurological deficits score in the different time as requested, use the digital camera record;
(4) experimental animal with equal conditions compares group, and except that occluding vascular not, all the other operation processs, record, scoring process are the same.
Above-mentioned special its structure of microsphere ductus bursae is:
Main body is one section PE10 silica gel tube that adapts to operation needs length, its internal diameter 0.25 ± 0.05mm, the stainless steel silk of insertion diameter 0.1 ± 0.02mm in the external diameter 0.60 ± 0.05mm, PE10 silica gel tube;
Described PE10 silica gel tube one end and 1 milliliter of syringe needle sealing socket; The other end of PE10 silica gel tube is provided with the silica gel-latex beads capsule of sealing, naturalness external diameter 1.0 ± 0.01mm during the unaerated of microsphere capsule;
The total length of described stainless steel silk is less than the PE10 length of tube, and the one end remains in the syringe;
Described PE10 silica gel tube outside tube wall mark length scale; Sacculus is partly immersed preservation, standby in the heparin sodium aqua.
Above-mentioned method is only blocked the blood in one-sided middle cerebral artery blood supply district, does not influence anterior cerebral artery, posterior cerebral artery.
A kind of " making the methods and applications of focal cerebral ischemia disease model with macaque ", application in human focal cerebral ischemia disease research and treatment, it is characterized in that the cerebral thrombosis that it can simulating human, cerebral infarction, or the sings and symptoms of transient ischemic attack, for scientific research and clinical treatment provide necessary primate cerebral ischemia disease model.
Compared with the prior art, advantage of the present invention is:
In the prior art operation process other model institute is unavoidable often to the infringement of blood vessel itself, even on apparatus, coat latex as the folder method of closing, also often can not be from addressing this problem in essence.
And the microsphere ductus bursae that the present invention adopts is less, gets involved in the middle cerebral artery only 2h behind the balloon expandable, and is therefore, less to the damage that blood vessel itself causes.
2. though the sub-method of vascular peg stay can overcome outside blood vessel the deficiency that folder closes methods such as middle cerebral artery, but can not irritate again, the perhaps uncontrollable concrete time of irritating again, in fact, the situation that this blood vessel is no longer logical is rare clinically, general cerebral thrombosis revascularization after thromboembolism treatment or the spontaneous dissolving of thrombosis, and the microsphere ductus bursae that our method is used, but occluding vascular during inflation, bleed off the revascularization that gas just can make obstruction, operate very easyly, can arbitrarily control time of cerebral ischemia fully, thereby can control the degree of ischemia, for the experimentation focal cerebral ischemia with irritate again proper model is provided.In fact, if make sacculus keep inflated condition always, can simulate permanent occlusion.
3. only in operation on neck operation, littler to the damage of brain than operation of opening cranium, guaranteed the integrity of cranial cavity to have reduced influence to the extra operation of cerebral ischemic model; Simultaneously, during operation on neck, use local anesthesia, therefore, the amount ratio routine of general anesthesia medicine is significantly reduced, reduced the influence of medicine the systemic arterial blood pressure.Because cranial cavity kept integrity, for the nervous physiology of long period and pathological observation provide may.
In a word, macaque blood vessel middle cerebral artery interpolation in a subtle way foley's tube make it to block or irritate again and can form the nervous system performance similar to people's syndrome similar to wind disease.The time of cerebral ischemia can be controlled, and is easy and simple to handle, at the research focal cerebral ischemia and have tangible advantage aspect the filling again; And except that macaque, also may be used for other larger animal.Therefore, has certain application prospect.
Description of drawings
Fig. 1 uses the present invention and laboratory animal is caused the situation that causes the left limb hemiplegia behind the middle cerebral artery occlusion of right side;
Fig. 2 uses the present invention and causes middle cerebral artery front and back, obstruction right side local cerebral blood flow to change to laboratory animal;
Fig. 3 uses the present invention laboratory animal is caused 3 hours magnetic resonance T2 of cerebral ischemia and blood vessel imaging;
Fig. 4 uses the present invention laboratory animal is caused remaining cell in the ischemia striatum (hematoxylin-Yin's red colouring, * 200 times) microphotograph;
Fig. 5 apparatus of the present invention, microsphere ductus bursae form sketch map (the most advanced and sophisticated sacculus that forms of inflation rear tube);
Fig. 6 apparatus of the present invention, the simulation sketch map of microsphere ductus bursae blocking blood flow (inflation back formation in the conduit reeve pipe is blocked).
The specific embodiment
Fig. 1 shows: middle cerebral artery occlusion/irritate the again typical function of nervous system defective that is caused is similar with the mankind's apoplexy.Main focal sign is that hemiplegia and eyeball are stared to Ipsilateral.And these signs can occur after generally blocking middle cerebral artery, and the beginning ocular bobbing is just fixing then and stare to Ipsilateral.
The result bradykinesia at first occurs after showing all animal right side middle cerebral artery occlusions, drowsiness even stupor; The left limb activity is ineffective, muscular strength reduces so that paralysis.These signs increase the weight of with the prolongation of ischemic time, 31.22 ± 11.09 minutes of function of nervous system scoring being reduced to when irritating 23h again in 38.75 ± 10.31 minutes when irritating 3 h again.And the sham operated rats that does not have occluding vascular is not seen this tangible sign.P＜0.05, statistical procedures has significant difference.
Fig. 2 shows: observe caudatum head local cerebral blood flow and begin rapidly to descend behind middle cerebral artery occlusion in experiment, about 2h touches the bottom; Irritate the back local cerebral blood flow again and go up gradually, but do not go back up to the preceding level of experiment.The local cerebral blood flow of sham operated rats also has certain fluctuation, but the scope that changes is little, generally about ± 15%, and unknown significance difference in the time period of laboratory observation.Cerebral ischemia group and sham-operation are compared, and 1h behind obstruction, 1.5h and 2h have significant difference (p＜0.05).
Fig. 3 shows: middle cerebral artery occlusion 2h, and the iconography that conforming to sign all appears in all animals changes, and tangible cerebral cortex and basal ganglia region infraction appear in " morbidity " 5h.The sign of irritating the 22h cerebral ischemia again is more obvious, the hemisphere swelling of tangible right side occurs, and the brain center line moves to left, even cerebral hernia occurs; The performance degree of MRI is consistent with the behavior of animal performance.Fig. 3 shows the brain nuclear magnetic resonance image and the corresponding brain vessel blood imaging of irritating 3h again.The cerebral infarction that nuclear magnetic resonance, NMR T2 picture shows is a high density area, mainly is distributed in brain cortical areas; The interior blood flow of middle cerebral artery that shows bilateral among the figure is equal substantially, and prompting blood vessel interpolation is gone into foley's tube and kept the limited pressure of certain hour blood vessel itself not to be caused tangible infringement.
Annotate: left side figure is a Magnetic Resonance Angiography, and it is uninfluenced that arrow shows that conduit is pulled out back blood vessel itself, and blood flow is normal.
Fig. 4 shows: middle cerebral artery occlusion/neuro pathology's change of typical cerebral ischemia appears in 24h macaque brain after irritating again.Because the brain specimen needs to carry out the necessary piece of repairing in the process of making section, what obtain is not the serial section of full brain, therefore it may be inappropriate calculating infarct volume according to infarct size, so adopt specific tangent plane to carry out infarct size relatively, can reflect the difference of cerebral ischemia degree as common aspect; Select for use the A17.5 aspect can see structures such as the cortex of ischemia, striatum according to the monkey brain collection of illustrative plates.The result shows that in the A17.5 aspect, cerebral infarct size is 59.197 ± 12.869mm
2Simultaneously can also see the obvious swelling of right side cerebral hemisphere, compare with sham operated rats, obviously increase of swelling coefficient (Ipsilateral hemisphere area/offside hemisphere area) (middle cerebral artery occlusion is irritated with sham operated rats again and is compared: 1.280 ± 0.017 to 1.016 ± 0.019, and p＜0.05).
The cortex and the striatal cellular morphology of microscopically offside (left side) are normal, and in the striatum center of ischemia side (right side) tangible neuron loss are arranged, and its morphosis of remaining cell is unusual; Damaged the obvious of striatum ischemic surrounding zone that be not so good as of the neuron of cortex ischemia surrounding zone.Fig. 4 shows that remaining cell is than the obvious minimizing of its offside (B) in impaired striatum center (A) district; And matched group does not find that the neuro pathology of cerebral ischemia changes.
The specific embodiment
Laboratory animal macaque (Macaca mulatta) is called yellow monkey, Rhesus Macacus, Guangxi monkey, belongs to monkey section; Be divided into experimental group (8 example) and matched group (or claim " sham operated rats ", 5 examples), every group is repeated different number of times according to requirement of experiment.Concrete condition is as follows:
13 experiment macaques, male and female are regardless of, 3～4.5 years old (be equivalent to teenager, jenny there is no menstruation, therefore can get rid of the influence of hormone and sexual cycle), body weight 4.35 ± 0.23Kg.Provide by Shanghai physiology institute of Chinese Academy of Sciences apery, confirm non-communicable disease through quarantine, can be for experiment.
Use the net laws on arrests during seizure, draw back the about 5cm of left and right sides active pulling-door in monkey cage front, with homemade monkey net (network interface 30 * 28cm, mesh 4 * 8cm, decreased food is 95cm) stretch in the monkey cage, cover monkey rapidly from a specific angle, the monkey net that overturns fast then takes it outside cage to.
Experimental animal is fixed in operating-table, the routine disinfection operative site, implement operation according to the following steps:
1. anesthesia: 8% chloral hydrate lumbar injection.Earlier with half amount (150mg/kg) injection; Anaesthetize with 1% procaine the part; Local a little 2% lignocaine that drips when handling carotid sinus.Owing to use local anaesthesia, the amount of general anesthetic is reduced than convention amount, thereby reduce the influence of medicine blood pressure.
2. operation: dorsal position, right shoulder bed hedgehopping.Preserved skin, the other improvement in cervical region right side median incision.Separate subcutaneous tissue, expose carotid sheath.Separate vagus nerve and common carotid artery, internal carotid artery with the glass minute hand) and external carotid artery.Separate ligation occipital artery, superior thyroid artery and arteria mandibularis.On external carotid artery, stamp standby knot.Close internal carotid artery and common carotid artery with the of short duration folder of bulldog clamp.
3. extract the 0.5ml arterial blood and do blood gas analysis.
4. cut off the external carotid artery distal end, insert in advance the microsphere ductus bursae that soaked with heparin (during unaerated from the external carotid artery stump, naturalness, external diameter φ 1.0 ± 0.01mm), insert about 59-62mm from internal carotid artery and external carotid artery crotch, stop the initial part that can block middle cerebral artery this moment when meeting obstructions.The ligation external carotid artery broken ends of fractured bone unclamps the bulldog clamp on common carotid artery and the internal carotid artery, makes middle cerebral artery blood supply district ischemia, at first emits gas behind the 2h, and the pumpback pintle keeps certain negative pressure, extracts the microsphere ductus bursae under the local anaesthesia out, and ischemic region is irritated again.
5. after operation finished, animal was positioned in 25 ℃ of air-conditioned rooms and rests.
Carry out neurological deficits score in the different time as requested, use the digital camera record.The postoperative animal gives different disposal.
" matched group " (or pressing the conventional jargon of medical experiment " sham operated rats "), except that occluding vascular not, all the other operation processs, record, scoring process are the same.
6. local cerebral blood flow monitoring: the experiment that will meet the requirement of macaque brain map body weight is fixed on Narishige SN-3 type stereo brain orienting instrument with monkey, position (the A18.0mm that on the macaque skull, is equivalent to the RCN head end, R8.0mm) with the saturating skull of the vertical brill of miniature electric drill, the trocar sheath of local cerebral blood flow monitoring probe is slowly inserted predetermined position, insert the local cerebral blood flow laser probe.Use PERIFLUX5010 program record, the result deposits computer in.With preceding 2 the local cerebral blood flow numerical value of cerebral ischemia is that benchmark calculates the shared percentage ratio of different local cerebral blood flow numerical value constantly as index.
7. behavior observation and function of nervous system infringement scoring: in monkey test chair, carry out.Lure it to dehisce with Fructus Musae, observation has or not distortion of commissure; Stimulate two upper limb, test muscular strength, muscular tension.Used grade form mainly draws certainly: Spetzler etc. " behavior observation and function of nervous system's infringement grade form " (Spetzler RF, Selman WR, Weinstein P, Townsend J, Mehdorn M, Telles D, Crumrine RC, and Macko R.Chronic reversible cerebral ischemia:Evaluation of a new baboonmodel.Neurosurgery, 1980,7:257-261), wherein animal dead is 0 minute, normally is 100 minutes.Comprise: consciousness situation, motor capacity and eye and cranial nerve function.
8. nuclear magnetic resonance scanning and blood vessel imaging adopt Siemens 63-SP4000 superconduct magnetic resonance device, magnetic field intensity 1.5 spies (tesla), and gradient field intensity 10mT/m uses the standard quadrature coil.Use self-rotary echo-pulse series, bed thickness 3mm obtains crown position, sagittal plain and level position T
1The weighting picture need be known (T
1Weighted imaging, T
1WI), T
2Weighting picture (T
2Weightedimaging, T
2WI), and proton density weighting picture (proton density weighted imaging, PDWI).(magnetic resonance angiography MRA) carries out blood flow imaging to adopt magnetic resonance angiography.All the other animals are in irritating 3h again except that the intact animal, 7h, and 22h scans, and analyzes T
1Weighting picture and T
2Signal intensity on the weighting picture.
9. the section of cerebral tissue and hematoxylin-Yin's red colouring (H﹠amp; E dyeing, hematoxylin and eosin stain): after macaque ischemia 2h irritated 24h again, left ventricular cannulation was got brain after perfusion is fixing, place 4 ℃ to contain 20% sucrose+4% paraformaldehyde solution to sinking, move into then in 4 ℃ of fixatives that contain 30% sucrose again to sinking.The continuous coronal paraffin section, thickness 6 μ choose 1 every 20 sections.Paraffin section scalds sheet through water-bath, is attached on the pretreated microscope slide.56 ℃ of 1h, 37 ℃ of baking ovens spend the night, and 4 ℃ of refrigerators are preserved.H﹠amp; E dyeing.The neutral gum mounting.Optical microscope is observed down.
10. cerebral infarct size and remaining cell rate calculate: according to macaque brain map coronalplane A17.5 plane, calculate cerebral infarct size, full brain sheet area and offside hemisphere area.Respectively the cortex of ischemia side and control sides (ischemia offside) and striatum are carried out the cell numeration under the light microscopic, method is as follows: demarcate institute's favored area, find index point under mirror, three visuals field of picked at random around it, to visual field inner cell counting, get three visual field meansigma methodss.Calculate the method for remaining cell percentage: ischemia side cell counting accounts for the Cytometric percentage rate of its ischemia offside.
11. Data Processing in Experiment and statistical analysis total data represent that with mean ± standard deviation each time point of brain electricity is relatively handled with variance analysis, each organizes cerebral infarct size and the cell survival rate is relatively checked with t.
About self-control microsphere ductus bursae
Because opening cranium middle cerebral artery occlusion/again irritates and is easy to form vasospasm through eye socket method occluding vascular, when the sub-method of vascular peg stay is uncontrollable again irritates again and can irritate on earth again, therefore, we utilize the sub-method of vascular peg stay directly blood vessel not to be caused the advantage of damage, overcome the shortcoming of embolus method, replace embolus with inflatable balloon, can control the time and the degree of cerebral ischemia arbitrarily.
Can't satisfy experimental requirements on the foley's tube specifications and characteristics of Xiao Shouing in the market, so need self-control.This cover foley's tube is made up of a small sacculus and the fine duct and the syringe that are attached thereto.The gas that injects trace by syringe makes sacculus full, thereby blocks endovascular blood flow, takes out sacculus reduction behind the gas, makes in the blood vessel blood flow logical again.Foley's tube technical parameter design according to macaque cerebrovascular anatomy (Macdonald RL, Kowalczuk A, OhnsL.Emboli enter penetrating arteries of monkey brain in relation to their size.Stroke.1995,26 (7): 1247-50) show, the diameter of the monkey brain middle cerebral artery The initial segment of crossing through formalin fixed is 0.7～1.1mm, in order to block the trunk of live body macaque middle cerebral artery, when body weight 2.8～4.7Kg, obturator diameter and artery diameter ratio should be 1.5: 1.(Molinari GF such as Molinari, Mossley JI and Laurent JP.Segmental middle cerebral artery occlusion inprimates:An experimental method requiring minimal surgery and anesthesia.Stroke 1974 is that successfully to block middle cerebral artery be exactly good evidence for the silica gel embolus of 1.6mm with diameter 5:334-339).Postmortem proves that the diameter of fixed preparation middle cerebral artery is 1.0mm.The variation of macaque MCA diameter is very little, mutual no dependence in this body weight and age bracket, and the thickness of internal carotid artery is relevant with body weight and age.Therefore, in selected the weight of animals and the range of age, the conduit of selecting diameter 1.0～1.1mm for use is valid, and the intubate resistance is very little during experiment.
To the about 3.0～3.5mm of the petrous bone of internal carotid artery, and petrous bone is 2.8～3.0mm to the MCA The initial segment from internal carotid artery, external carotid artery branching.Macaque cerebrovascular brain Willis ring has unique distinction: middle cerebral artery and anterior cerebral artery angle are bigger, anterior cerebral artery almost is horizontal traveling, therefore conduit directly enters middle cerebral artery easily, rather than the cerebrovascular of rat, enters anterior cerebral artery during plug wire easily.Therefore the foley's tube of calibrated length is imported and to feel to have resistance when blood vessel arrives corresponding scale.
In addition, " S " type vascular bending often caused difficult intubation after conduit entered petrous bone, and therefore, catheter tip does not have stainless steel wire inner core when making foley's tube, thereby makes catheter tip soft; If action is easy to cause vasospasm roughly during intubate, therefore, should not be slotting firmly repeatedly, intermittently continue again a moment, generally the elasticity by conduit can spring into conduit; If repeated multiple times can not be inserted, but inject small amount of lidocaine in the blood vessel.Foley's tube manufacturing process is summarized as follows:
1. the stainless steel silk that with diameter is 0.21mm inserts internal diameter 0.28mm, external diameter 0.61mm PE10 silica gel tube (commercial polyethylene medical silicone tube, trade mark PE10, factory: U.S. Davis ﹠amp; Geck company, Sherwood, NY, USA) in.
2. syringe is connected with the PE10 seal of tube, an end of stainless steel silk remains in the syringe, and the other end is than the short 7～8mm of PE10 pipe.
3. the tip of PE10 pipe is sealed with special glue, the other end and syringe are tightly connected, and occur siphonage when preventing impregnation.
4. the tip of PE10 pipe is inserted in the silica gel-latex mixed liquor according to the special ratios configuration, take out behind the 2-3sec.
5. 10min. in 37 ℃ of baking ovens
6. inject 0.01～0.02ml gas, form sacculus.
7. the sacculus gluing is reinforced.
8. dry naturally, fully solidify the back and check sacculus, venting back diameter can use at 1.0～1.1mm.
9. reference mark, signalment is gone into cranium place and sella turcica place, is respectively 3.0～3.5mm in this experiment, 6.0～6.5mm
10. sacculus is partly immersed in the heparin sodium aqua and preserve.
Vitro examination shows that sacculus still kept certain firmness at least under the room temperature in 3 months.
Note before using keeping balloon diameter 3.0～3.5mm in distilled water (twice distilled water) build-in test whole system, balloon diameter is constant in 12 hours can use; If gas leakage can not be used, the constant pressure when guaranteeing occluding vascular, otherwise can can't determine time of middle cerebral artery occlusion/irritate again.
Claims (5)
1. make the method for the disease model of focal cerebral ischemia with macaque for one kind, it is characterized in that:
Laboratory animal is chosen primate macaque Macaca mulatta;
Get involved the microsphere ductus bursae in the blood vessel, the initial part of blocking middle cerebral artery makes middle cerebral artery blood supply district ischemia by requirement of experiment, makes the cerebral ischemia disease model.
2. according to the described method of claim 1, it is characterized in that intervention microsphere ductus bursae method is in the described blood vessel with macaque manufacturing focal cerebral ischemia disease model:
(1) operation gets involved sacculus obstruction laboratory animal middle cerebral artery blood supply district
Insert the special microsphere ductus bursae that soaked with heparin in advance from the macaque external carotid artery, insert middle cerebral artery through internal carotid artery, be inflated to the full initial part of blocking middle cerebral artery of microsphere capsule to the microsphere ductus bursae, make middle cerebral artery blood supply district ischemia continue 2h, make the cerebral ischemia disease model;
(2) ischemic region is irritated again
By the interval of requirement of experiment, emit gas from the microsphere ductus bursae, extract the microsphere ductus bursae under the local anaesthesia out, ischemic region is irritated again;
Operation finishes, and after stitching was handled, laboratory animal was positioned in 25 ℃ of air-conditioned rooms and rests;
(3) record, scoring
Carry out neurological deficits score in the different time as requested, use the digital camera record;
(4) experimental animal with equal conditions compares group, and except that occluding vascular not, all the other operation processs, record, scoring process are the same.
3. according to claim 1 or 2 described methods, it is characterized in that described special its structure of microsphere ductus bursae is with macaque manufacturing focal cerebral ischemia disease model:
Main body is one section PE10 silica gel tube that adapts to operation needs length, its internal diameter 0.25 ± 0.05mm, the stainless steel silk of insertion diameter 0.1 ± 0.02mm in the external diameter 0.60 ± 0.05mm, PE10 silica gel tube;
Described PE10 silica gel tube one end and 1 milliliter of syringe needle sealing socket; The other end of PE10 silica gel tube is provided with the silica gel-latex beads capsule of sealing, microsphere capsule naturalness external diameter 1.0 ± 0.01mm;
The total length of described stainless steel silk is less than the PE10 length of tube, and the one end remains in the syringe;
Described PE10 silica gel tube outside tube wall mark length scale; Sacculus is partly immersed preservation, standby in the heparin sodium aqua.
4. according to the described method of claim 1, it is characterized in that described method is only blocked the blood in one-sided middle cerebral artery blood supply district, does not influence anterior cerebral artery, posterior cerebral artery with macaque manufacturing focal cerebral ischemia disease model.
5. the application in human focal cerebral ischemia disease research and treatment in accordance with the method for claim 1, it is characterized in that the cerebral thrombosis that it can simulating human, cerebral infarction, or the sings and symptoms of transient ischemic attack, for scientific research and clinical treatment provide necessary primate cerebral ischemia disease model.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100372523C (en) * | 2006-05-23 | 2008-03-05 | 天津郁美净集团有限公司 | Quickly absorptive skin-care composition |
| CN109172032A (en) * | 2018-10-13 | 2019-01-11 | 刘劼 | A kind of microballoon pipe line pin device of MCAO models in rats |
| CN109549739A (en) * | 2017-12-25 | 2019-04-02 | 温州医科大学附属第医院 | A kind of kit gathering cerebral ischemia animal model for constructing novel magnetic particle |
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2004
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100372523C (en) * | 2006-05-23 | 2008-03-05 | 天津郁美净集团有限公司 | Quickly absorptive skin-care composition |
| CN109549739A (en) * | 2017-12-25 | 2019-04-02 | 温州医科大学附属第医院 | A kind of kit gathering cerebral ischemia animal model for constructing novel magnetic particle |
| CN109172032A (en) * | 2018-10-13 | 2019-01-11 | 刘劼 | A kind of microballoon pipe line pin device of MCAO models in rats |
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