CN101653378A - Method for establishing artery ischemia model of brain of rhesus monkey and evaluation method - Google Patents
Method for establishing artery ischemia model of brain of rhesus monkey and evaluation method Download PDFInfo
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Abstract
The invention relates to a method for establishing an artery ischemia model of a brain of a rhesus monkey and an evaluation method. The constructing method adopts the mode of directly puncturing femoral arteries of an experimental monkey to perform the intravascular interventional operation under the direct view of a digital subtraction angiography, and establishes the chronic cerebral infarctionprimate model by using a micro-catheter to block the arteries in the brain of the experimental monkey and controlling ischemia time. The model is a quite reasonable cerebral ischemia-reperfusion animal model, and the pathological characteristics of the monkey cerebral injury model established by the method are similar to clinical manifestation; and moreover, the model is stable and reliable and issuitable for all kinds of curing therapeutic research and related basic research.
Description
Technical field
The present invention relates to a kind of method that makes up and assess Rhesus Macacus middle cerebral artery ischemia model, belong to field of medicaments.
Background technology
The cerebral infarction animal model plays important effect in the various researchs of ischemic cerebrovascular, especially aspect the assessment of various Therapeutic Method, as new drug evaluation etc.The animal that is used for this respect research at present comprises rat, rabbit, Canis familiaris L., cat and primate such as macaque, baboon etc.Meiofauna such as rat, rabbit uses comparatively extensive because of economy, simple operation and other advantages.But the species variation of these animals and human beingses is big, and the result and the clinical experiment that cause zoopery to obtain are not inconsistent.Monkey is very approaching with the people on kind, and its sensorimotor function all has similarity with the people.The distribution of monkey brain tissue morphology, neuron and neurogliocyte, cerebrovascular anatomical structure etc. all are similar to (Hardev Ramandeep SinghGirn, Reperfusion Syndrome:Cellular Mechanisms of Microvascular Dysfunctionand Potential Therapeutic Strategies.Vasc Endovascular Surg 2007 with the mankind very much; 41; 277.).The physiology and the clinical observation of the long term repeatability that symon etc. carry out show, the single arterial occlusion model of monkey is optimal cerebral ischemic model (Benjamin S.Geisler, et al.Blood OxygenLevel-Dependent MRI Allows Metabolic Description of Tissue at Risk in AcuteStroke Patients.Stroke, 2006,37:1778-1784.).Performance behind the monkey brain ischemia and people have a lot of similarities, myasthenia of limbs on the side can occur, can involve lower limb, hemianopsia etc. in the same way also can occur.Then often be difficult to estimate this class symptom at some meiofaunas.
Stable cerebral ischemic model should possess following characteristics: 1. can optionally inaccessible a certain tremulous pulse, and good reproducibility; 2. arterial occlusion can cause that part or a certain regional cerebral blood flow change, and induces brain essence infarction; 3. Bi Sai tremulous pulse can pass to again and guarantee higher animal dis motility rate.
Though the non-human primate cerebral ischemic model is the cerebrovascular disease of simulating human preferably, but because the source of animal is limited, cost an arm and a leg, use the cerebral ischemia experimental model of high animal structures such as monkey less relatively, still do not have standardized monkey brain ischemia model at present.Therefore, set up simple, good reproducibility, mortality rate is low and the monkey brain ischemia model that is easy to control Ischemia Time is to be worth the problem further explored.Monkey middle cerebral artery cerebral infarction models mainly contains two kinds of construction methods at present: a kind of is the intra-arterial embolization, and another kind is the method for closing the MCA initial part through the operation on orbit folder.As: Tan Longwang etc., the foundation of feeling anxious property cerebral infarction model, has reported that employing gelfoam embolization monkey internal carotid artery duplicates the acute cerebral infarction model at " southwestern medical officer " 2005 75 phases of volume.Concrete grammar is: select 6 of Adult Monkey for use, and the femoral artery puncture intubate, the gelfoam embolization internal carotid artery, line number word subtractive angiography (DSA) shows the vascular occlusion situation then, behind the bolt 3,4,6,8,12,24hCT is unenhanced.Observe cerebral infarction CT performance.As a result behind 6 monkey thromboembolisms DSA show in the neck moving inaccessible, before the brain, medium-sized artery do not develop.The unenhanced visible thromboembolism side cerebral hemisphere lamellar low-density of 6～8hCT dead band behind the ischemia.It uses the gelatin embolism internal carotid artery; And the supervision of no contrast apparatus in the operation.Cause be a side brain before, the permanent ischemia of medium-sized artery, the monkey mortality rate will be very high; And ishemic part may be also non-constant.Wei Riming etc., 2006 23 3 phases of volume of the foundation of local cerebral ischemia machin animal model " Guangxi Medical University's journal ", select the machin of 10 health, adopt blood vessel to get involved method from femoral arteriography, select to middle cerebral artery (middlecerebral artery microtubular is super, MCA) M1 branch injects the lean type thrombosis and stops up its blood vessel, blocks its blood flow.Result: behind the thromboembolism, by digitized video angioradiographic system (DSA) radiography MCAM1 branch vessel poor visualization; The visible acute local cerebral ischemia focus of CT Perfusion Imaging behind the thromboembolism 1h; Carry out the unenhanced visible low-density old local cerebral ischemia focus of CT behind the 48h.2 routine corpse are dissected, take out cerebral tissue TTC dyeing and be positive; All show in various degree symptoms of cerebral ischemia and sign after all the other animals are clear-headed.Conclusion: select MCA lean type white thrombus can set up stable monkey local cerebral ischemia model with super, because of this animal model is at present near the model of clinical case, research, diagnosis and treatment, particularly thromboembolism treatment to the generation development mechanism of apoplexy have practical preclinical phase using value.This animal model is nonvolatil ischemia model by the thromboembolism middle cerebral artery, and animal dead rate height is not easy to carry out drug screening.Feng Feiling etc., blood vessel intervention method imports and sets up monkey local cerebral ischemia model from the body thrombosis, " Chinese Journal of Pathophysiology " 2006 22 10 phases of volume, the document has reported that employing blood vessel intervention method imports from the body thrombosis, set up monkey local cerebral ischemia animal model, for the correlational study of cerebral ischemia provides the basic research that wound is little, application prospect is big platform. method: with 12 machins is experimental subject, adopt blood vessel intervention method microtubular to be surpassed and select branch, import from the body thrombosis and stop up MCA to middle cerebral artery (MCA) M1 from femoral artery.By the thromboembolism middle cerebral artery, be nonvolatil ischemia model, animal dead rate height is not easy to carry out drug screening.
Number of patent application: 200410075763.9, denomination of invention: the methods and applications of making the focal cerebral ischemia disease model with macaque, 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.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.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; This patent is carotid artery place operation separating blood vessel, and the blind method in back gets involved with sacculus and stops up middle cerebral artery, and does not have the contrast apparatus guiding.This class model operation wound is bigger, and thromboembolism can not accurately stop up a certain position of middle cerebral artery entirely with operator's experience but not look at straight under radiography, causes model ischemia effect difference very big probably, is unfavorable for further research.
But these methods all respectively are being weak aspect operability, concordance and the verity of anthropomorphic dummy's cerebral ischemia pathophysiological process.
Summary of the invention
The technical scheme that the invention provides has provided a kind of method that makes up and assess Rhesus Macacus middle cerebral artery ischemia model.
The invention provides a kind of method that makes up Rhesus Macacus middle cerebral artery ischemia model, it is to adopt directly to experimental monkey groups row femoral artery puncture mode, under the direct-view of Digital Subtraction machine, carry out interventional procedure in the blood vessel, block the experimental monkey groups middle cerebral artery and control the method that Ischemia Time is set up chronic brain infraction primates model with microtubular.
It specifically comprises the steps:
The anesthesia of a, animal: animal is prohibited drink fasting 10h before the anesthesia; 30min before the anesthesia gives atropine (0.05mg/Kg) intramuscular injection; With 3% pentobarbital sodium (1ml/Kg) intravenous injection, satisfied until anesthesia;
B, blood vessel and cerebral angiography: animal is got in operating-table and lies on the back, and is fixing, the conventional preserved skin in territory, right inguinal region, sterilization; Choosing the right femoral artery point of maximum intensity of beating is the capable blind method puncture of point of puncture; Insert 5F arteries sheath after the success, insert 4F angiography catheter (Cordis) under seal wire (Cordis) guiding, last blood vessel of row aorta bow and cerebral angiography are to understand entocranial artery situation out of shape;
After c, radiography finished, tail end of conduit connect Y type band valve union, and its side arm band threeway hose connector also links to each other with the pressure transfusion bag, continues to splash into normal saline in case thrombosis (4ml/min); Under DSA (Philip Allura Xper FD20) monitors, angiography catheter is inserted to below the internal carotid artery initial part, band is supported the 1.9F microtubular (Prowler-10 of seal wire; Cordis) send into a straight side MCA and arrive the M1 The initial segment to block middle cerebral artery until its head end; Block successfully and block position MCA far-end branch development situation later on by 4F conduit and the affirmation of 1.9F microtubular radiography respectively;
After d, the affirmation MCA blocking-up, the locking Y-valve, fixedly microtubular is located in this, causes lasting MCA to block; Withdraw from microtubular, conduit behind the thromboembolism 2h, pull out the arteries sheath, make the MCA blood flow logical again, form the Focal Cerebral Ischemia Reperfusion model.
The present invention also provides a kind of Rhesus Macacus middle cerebral artery ischemia model, and it is to be made up by said method to obtain.
The invention provides a kind of method of assessing Rhesus Macacus middle cerebral artery ischemia model, it is to go up the assessment of adopting based on the capable model of magnetic resonance method of dispersion tensor imaging and blood oxygen level dependence at 3.0T magnetic resonance imager (GE Signa-excite).
The present invention adopts the microtubular thromboembolism, carries out the modeling of blood vessel interventional procedure method under the direct-view of Digital Subtraction machine.Therefore this method has more advantage than other modeling methods, comprising: (1) can cause single arterial thrombosis, does not influence the formation of collateral circulation, more near the practical situation of people's cerebral ischemia; (2) the thromboembolism position is constant, and effect of embolization is reliable, thereby the controllability of the ischemia scope of model and degree of ischemia is better; (3) need not to use sacculus/microcoils, reduce cost, simplify the operation, and because do not produce pseudo-shadow, make it possible in cerebral ischemia super acute stage, carry out the assessment of the capable model of magnetic resonance imaging.(4) the animal dead rate is low, is suitable for setting up the chronic model that is used for the medicine assessment.
Femoral artery puncture, Digital Subtraction machine are looked at descending microtubular straight and are blocked primates cerebral infarction model ischemia effect sure, favorable repeatability, well the Pathophysiology situation of simulating human cerebral ischemia that experimental monkey groups middle cerebral artery mode is set up; The functional MRI method is responsive and well relevant with neurological symptom to the detection of cerebral ischemia; Both combinations are for the medicine that screens new treatment acute cerebral infarction provides a kind of new method; As seen this model is a kind of more satisfactory cerebral ischemia re-pouring animal model, and approximate with the lesion characteristic and the clinical manifestation of the duplicated monkey brain damage model of this method, model stability is reliable, is suitable for various therapeutic studies and relevant basic research.
Description of drawings
Fig. 1 is cerebral angiography result before and after the animal MCAO-003 RMCA thromboembolism.Fig. 1, A be before the RMCA thromboembolism through the capable right side of 4F conduit internal carotid artery radiography result, that visible RMCA develops is good (among the A figure shown in the arrow).Fig. 1, B are the RMCA thromboembolism after the capable right side of 4F conduit internal carotid artery radiography result, visible RMCA trunk do not develop (among the B figure shown in the black arrow); MCA M1 section near-end branch develop (among the B figure shown in the white arrow).Fig. 1, C are the RMCA thromboembolism after the capable super radiography result that selects of microtubular, and the head end labelling of visible microtubular is arranged in M1 section far-end (shown in the C figure arrow); Also show M1 section far-end blood simultaneously for situation, visible M2 section and distal vessels thereof are developed good.
Fig. 2. be the MRA image of animal MCAO-003.Fig. 2, A are 1h MRA image behind the ischemia, and visible RMCA blood flow signal disappears; Fig. 2, B and Fig. 2, C are respectively 2h and postoperative 24h MRA result behind the ischemia, and visible RMCA blood flow signal partly recovers (among the figure shown in the arrow), and apparent infarction kitchen range is arranged.
Fig. 3 is 4 the continuous layer images (axle position) of 1h animal MCAO-004 behind the ischemia.Fig. 3, A are ADC figure, and a plurality of aspects are involved in large stretch of low signal district in the visible RMCA blood supply district, are the ischemia focus; Fig. 3, B are T2 FLAIR image, and abnormal signal does not appear in this fashion.
Fig. 4 is 4 continuous sweep layer images (axle position) of perfusion back 2h animal MCAO-004 again.Fig. 4, A are ADC figure, unusual low signal focus in the still visible RMCA blood supply district; Fig. 4, B are T2FLAIR, visible unusual high signal focus, and its distribution is consistent with RMCA blood supply district.
The relative average surface dispersion coefficient of Fig. 5 is change curve in time: the rADC value in visible several zones all reduces behind ischemia, and prolongs in time and descend.Its value of perfusion back prolongs in time and rises again.
The relative average surface dispersion coefficient of Fig. 6 is with the change in location curve: as seen near more low more apart from the ischemia central area in different time point rADC values, and far away more high more apart from the ischemia central area, there is linear relationship roughly.
The relative anisotropic mark of Fig. 7 is with the change in location curve: the trend of having represented the rAF value.The value that is ischemia half blanking bar zone (2 district) significantly raises behind ischemia.And experiencing a normalization process after the perfusion again, then raising once more.
Fig. 8 blood oxygenation level relies on signal with the change in location curve: represented the degree that T2 ' reduces than offside and the relation of infarcted region.The near more T2 ' of visibility infarction kitchen range reduces many more.And infarction core space T2 ' changed in time rise not obviously, and double blanking bar zone and optimum ischemic region T2 ' reduction value have the trend of minimizing gradually.
The specific embodiment
Set up a cover makes up Rhesus Macacus middle cerebral artery ischemia model and functional MRI assessment institute established model by the intra-arterial interventional method rules.
Test material
1, main agents
Anesthetis: 3% pentobarbital 30mg/kg
Atropine sulfate 10mg/2ml
Disinfectant: polyvidone povidone iodine 5.0g/L
Get involved contrast agent: Iopromide injection 15g/50ml
Magnetic resonance developing agent: the gadolinium spray amine 0.5mol/L of Portugal
2, key instrument, apparatus
The key instrument equipment introduction;
Philip Allura Xper FD20 angioradiographic system
Cordis Prowler-10 microtubular, Cordis 4F angiography catheter, Cordis guides seal wire
GE Signa-excite 3.0T superconduct magnetic resonance imaging system
The special-purpose two-tube high pressure injector of Medrad magnetic resonance
RM6240BD animal electrocardiogram monitor (Chengdu Instruement Factory)
Sphygomanometer, ear temperature meter
3, animal and feeding and management
3 Rhesus Macacus, the weight of animals scope 5-9kg, the range of age 5-8 year, male.
Rhesus Macacus is raised in conventional animal room, Chinese medicine safety evaluatio center, national Chengdu, the preceding animal of experiment conforms and carries out the anthelmintic treatment, select healthy (female palpus unpregnancy) animal as animal subject, rustless steel monkey cage sub-cage rearing, the raising condition meets GB GB14925-2001,18 ± 26 ℃ of room temperatures, relative humidity 40 ± 70%.Clear and mischievous person's forage feed, tap water bottling are freely drunk, every day feed fruit.
Animal is hung on neck board and the cage and labels as animal identification marking.
Test example 1 makes up the method for Rhesus Macacus middle cerebral artery ischemia model
1, the anesthesia of animal: animal is prohibited drink fasting 10h before the anesthesia; 30min before the anesthesia gives atropine (0.05mg/Kg) intramuscular injection; With 3% pentobarbital sodium (1ml/Kg) intravenous injection, satisfied until anesthesia;
2, Rhesus Macacus cerebral infarction model modeling operation method
2.1. animal different fasting 12h before the art.
2.2. (the Animal Anesthesia method is with reference to Rhesus Macacus surgery anesthesia SOP) all carried out in all modeling operations under the general anesthesia state.
2.3. animal is got in operating-table and lies on the back, and fixes with the self-control cystosepiment.With the binder fixing limbs.
2.4. the conventional preserved skin of right side inguinal region.
2.5. sterilization art district three times, scope surpasses 5 centimetres of operation puncturing points.Field of operation shop aseptic towel.
2.6. choosing the right femoral artery point of maximum intensity of beating is the capable blind method puncture of point of puncture.
2.7. insert 5F arteries sheath after the success, under seal wire guiding (Cordis), insert 4F angiography catheter (Cordis), the row aorta bow is gone up blood vessel and cerebral angiography to understand entocranial artery situation out of shape.
2.8. after radiography finished, tail end of conduit connect Y type band valve union, its side arm band threeway hose connector also links to each other with the pressure transfusion bag, continues to splash into normal saline in case thrombosis (4ml/min).Under DSA monitors, angiography catheter is inserted to the internal carotid artery initial part, then will be with the 1.9F microtubular (Prowler-10 that supports seal wire; Cordis) send into a straight side MCA and arrive the M1 section until its head end.
2.9. block successfully and block position MCA far-end branch development situation later on by 4F conduit and the affirmation of 1.9F microtubular radiography respectively.
2.10. after confirming the MCA blocking-up, the locking Y-valve, fixedly microtubular is located in this, causes lasting MCA to block.Get the cleaning adhesive plaster conduit and arteries sheath exposed parts are fixed in the animal thigh root, rapidly animal is transferred to the magnetic resonance examination chamber.
2.11. withdraw from microtubular, conduit behind the thromboembolism 2h, pull out the arteries sheath, make the MCA blood flow logical again, form the Focal Cerebral Ischemia Reperfusion model.
2.12. the body temperature of periodic monitor animal, breathing and blood pressure situation in the art.
3, iconography evaluation operation method
3.1. animal different fasting 12h before the iconography assessment.
3.2. (the Animal Anesthesia method is with reference to Rhesus Macacus surgery anesthesia SOP) all carried out in all iconography assessments under the general anesthesia state.
3.3. MRI scans with the eliminating intracranial lesion before the row art, and leaves and takes the baseline data.
3.3.1. animal lies on the back in the MRI scan table, pad with disposable plastic bag keeping clean, and lid with woollen blanket with warming.
3.3.2. head is positioned in the birdcage head coil, its head both sides of cotton pad filling is to guarantee that it is over against instrument.
3.3.3. row 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map, PWI, PWI-T1, MRA scan with the eliminating intracranial lesion, and adjust and set up each sequential parameter of Rhesus Macacus magnetic resonance imaging.
3.4. operation day is sterilized postanesthetic experimental monkey groups one side upper limb preserved skin, and with the special-purpose anti-high pressure needle row vein puncture of PWI and standby with heparin in sealing with the 5ml syringe.
3.5. MRI scanning in 1 hour behind the operation ischemia.
3.5.1. the animal allocation method is seen 3.3.1.
3.5.2. high pressure injector has been aspirated contrast agent, and has been connected with syringe needle.Occur beginning with 5ml/s speed injection contrast agent in first original perfusion image at the beginning sequence scanning.
3.5.3. row 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map, PWI, PWI-T1 scanning.
3.6. later scanning sequence is respectively:
Behind the ischemia 2 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map
Poured into again back 1 hour: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map, MRA
Poured into again back 2 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map
Poured into again back 3 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map, PWI, PWI-T1
Poured into again back 4 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map
Poured into again back 5 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map
Poured into again back 6 hours: 3PL, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map
Postoperative the 1st day: 3PL, DTI, DTI T1, T2
*Map, T2
*-T1, T2map, MRA
Postoperative the 7th day: 3PL, T2-FLAIR, DTI, DTI-T1, T2
*Map, T2
*-T1, T2map, MRA
Its concrete operation method is with 3.5.
4, neuroethology evaluation operation method
4.1 evaluation time
To the Rhesus Macacus of having quarantined before ischemia, behind the ischemia 12 hours, 24 hours, 48 hours, the scoring of the capable function of nervous system of same time point the 3rd, 4,5,6,7 day every day
4.2 evaluation personnel
Undertaken by two experiment members that participate in the concrete operations but the unknown is performed the operation of this experiment
4.3 evaluation methodology and standards of grading
Earlier Rhesus Macacus being carried out the daily life habit before the experiment observes, be familiar with its basic rule of life and behavioural habits, estimate state of consciousness, motor system, skeletal muscle coordination function and sensory system then in the list of references in the neuroethology evaluation methodology at primate or large animal cerebral ischemic model; The scoring form is as follows:
Neurobehavioral marking standard behind the monkey local cerebral ischemia
Annotate: amounting to best result is 100 minutes
Large animal function of nervous system grade form
5, iconography data processing method
5.1 parameter acquisition:
The DIT image reconstruction is gone out average dispersion coefficient (DCvag) figure, part anisotropy (AF) figure.The gained parameter comprises DCvag, AF value.
With T2
*Map and T2map functool2 software rebuild, and line function subtracts each other, and obtains T2 ' figure.
5.2 (ROI) interested distinguishes selection:
Two experimenters select 4 ROI with blind method, are 6.3mm (16pix).1 is the infarction center, and 3 is the marginal zone, and 2 is 1,3 mid point, and 4 is DTI normal signal district.Measure DCvag, AF, T2 ' value.For avoiding of the influence of infarction position difference, adopt the relative value of pathological changes side and normal offside during calculating to experimental result.
5.3 with the capable statistical analysis of spsssl3.0 software
Result of the test:
Rhesus Macacus middle cerebral artery ischemia modeling effect and evaluation
To number 03223 experimental monkey groups is example, and its iconography assessment result is as follows
1, digital subtraction angiography assessment result
Row thromboembolism side internal carotid artery radiography, as seen this side MCA trunk does not develop, and confirms microtubular thromboembolism MCA success, and at this moment, M1 section proximal part branch is still as seen; Through the super radiography of selecting of the capable MCA of microtubular, see the thromboembolism position with blood vessel development far away, and show microtubular head end position (being the thromboembolism position) then.(see figure 1)
2, nuclear magnetic resonance, NMR assessment result
2.1 common sequence MRI image evaluation result
Assessment MRI image, confirmation truly has the obturation of MCA, and the infarction kitchen range due to the ischemia.(see figure 2)
2.2 function sequence MRI image evaluation result
After the MCA obturation, the DTI of 3 animals has shown and the consistent ischemic injuries focus of thromboembolism side MCA blood supply district distribution with T2FLAIR result.1h behind the ischemia, and ADC figure display abnormality low signal focus (Fig. 3, A), its distribution is consistent with RMCA blood supply district; At this moment, the T2FLAIR picture do not occur as yet abnormal signal (Fig. 3, B).Perfusion back 2h again, ADC figure and T2FLAIR picture all show abnormal signal in the RMCA blood supply district (Fig. 4, A-B).
2.3 function sequential parameter assessment result
ADC, AF, 1-VR, RA, T2 ' value result and development law are as follows:
Table 1
The rADC value in the visible several zones of Fig. 5 all reduces behind ischemia, and prolongs in time and descend.Its value of perfusion back prolongs in time and rises again.Fig. 6 is as seen near more low more apart from the ischemia central area in different time point rADC values, and is far away more high more apart from the ischemia central area, has linear relationship roughly.Fig. 7 has represented the trend of rFA value.The value that is ischemia half blanking bar zone (2 district) significantly raises behind ischemia, and is experiencing a normalization process after the perfusion again, then raises once more.Fig. 8 has represented that T2 ' is than the degree of offside reduction and the relation of infarcted region.The near more T2 ' of visibility infarction kitchen range reduces many more.And infarction core space T2 ' changed in time rise not obviously, and double blanking bar zone and optimum ischemic region T2 ' reduction value have the trend of minimizing gradually.
ADC wherein: surface dispersion coefficient (apparent diffusion coefficient)
RADC: relative ADC (apparent surface's dispersion coefficient)
RFA: relative FA (infarcted region fractional anisotropy/offside normal brain activity district fractional anisotropy).
T2 ': the blood oxygenation level relies on signal
3, neuroethology assessment result
To number 03223 experimental monkey groups is example, and visible experimental monkey groups has the process of recovering gradually from function of nervous system's infringement that ischemia causes.
Neurobehavioral marking standard behind the monkey local cerebral ischemia
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Large animal function of nervous system grade form
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Animal model of the present invention can be used for screening treatment cerebral infarction medicine, and is also useful in research cerebral infarction mechanism.
Cerebral infarction sickness rate height, very harmful to human beings'health, because the pathogeny of primary disease is not exclusively definite, so clinical effective medicine and the treatment means of still lacking.And the hysteresis of experimental animal model research is one of bottleneck that the above pathogenic mechanism of restriction is studied and medicine is developed.Therefore, it is particularly important to the correlational study of cerebral infarction to seek suitable animal model.With the front subject that to substitute, repair or reproduce the various regenerative medicines that are organized as characteristic of human body are medical research field, increasing regenerative medicine product has been applied to clinical treatment.The anatomical features of primate, physiological function and very close to the reactivity and the mankind of disease and medicine are the responsive animals that carries out the clinical preceding efficiency evaluation of regenerative medicine product.
The Rhesus Macacus cerebral infarction model scope of application of this research and establishment is given an example:
1. the VEGF of discovered in recent years (vascular endothelial growthfactor, VEGF) be the short vascular endothelial cell proliferation factor of high special, be under the jurisdiction of neurotrophic factor family, participate in generation, the development of whole cerebral ischemia pathological process; Induce new vessels to generate, promote side Zhi Xunhuan to set up; Suppress neuronal apoptosis and the death of tardy property.Can utilize the special biological action of Rhesus Macacus cerebral infarction model research VEGF, to excavate new treatment means, for human treatment's cerebral ischemia diseases provides possible method.
2. scientist has poured into great amount of manpower and material resources in the research of cellular replacement therapy cerebrovascular disease.The correlational study of embryo stem cell transplantation treatment cerebral infarction is limit finally stranded because of ethics reason and rules.Recently (mesenchymal stem cell, study to interstital stem cell by transplanting MSC) with attention diversion for researcher.MSC has multiple differentiation potential, can be divided into osteoblast, chondrocyte, adipose cell, sarcoplast and neuronal cell, and proliferative ability is strong, and plasticity, animal migration and integration performance are good, and can see through blood brain barrier.Someone thinks that MSCs may be that (embryonic stem cells ESCs), has the characteristic of ESCs to reservation (hiding) embryonic stem cell in the adult tissue organ.The discovery of MSC and successfully be divided into neurocyte, for the transplantation treatment of nervous system disease provides " seed cell ", there is research to report that MSC transplantation treatment cerebral infarction can alleviate the rat neurologic impairment in the recent period, and stereotaxis transplanting, intravascular injection transplanting, subarachnoid space are transplanted to transplant all in (pillow Da Chi injection, waist are worn injection), the tricorn and are succeedd, and can utilize the Rhesus Macacus cerebral infarction model to study the mechanism of action and the therapeutical effect of this method.
Claims (5)
1, a kind of method that makes up Rhesus Macacus middle cerebral artery ischemia model, it is characterized in that: it is to adopt directly to experimental monkey groups row femoral artery puncture mode, under the direct-view of Digital Subtraction machine, carry out interventional procedure in the blood vessel, block the experimental monkey groups middle cerebral artery and control the method that Ischemia Time is set up chronic brain infraction primates model with microtubular.
2, the method for structure Rhesus Macacus middle cerebral artery ischemia model according to claim 1, it comprises the steps:
The anesthesia of a, animal: animal is prohibited drink fasting 10h before the anesthesia; 30min before the anesthesia gives atropine 0.05mg/Kg intramuscular injection; With 3% pentobarbital sodium 1ml/Kg intravenous injection, satisfied until anesthesia;
B, blood vessel and cerebral angiography: animal is got in operating-table and lies on the back, and is fixing, the conventional preserved skin in territory, right inguinal region, sterilization; Choosing the right femoral artery point of maximum intensity of beating is the capable blind method puncture of point of puncture; Insert 5F arteries sheath after the success, insert the 4F angiography catheter under the seal wire guiding, last blood vessel of row aorta bow and cerebral angiography are to understand entocranial artery situation out of shape;
After c, radiography finished, tail end of conduit connect Y type band valve union, and its side arm band threeway hose connector also links to each other with the pressure transfusion bag, continued to splash into normal saline in case thrombosis, and a speed is 4ml/min; Under DSA monitors, angiography catheter is inserted to below the internal carotid artery initial part, the 1.9F microtubular of band support seal wire is sent into a straight side MCA arrive the M1 The initial segment to block middle cerebral artery until its head end; Block successfully and block position MCA far-end branch development situation later on by 4F conduit and the affirmation of 1.9F microtubular radiography respectively;
After d, the affirmation MCA blocking-up, the locking Y-valve, fixedly microtubular is located in this, causes lasting MCA to block; Withdraw from microtubular, conduit behind the thromboembolism 2h, pull out the arteries sheath, make the MCA blood flow logical again, form the Focal Cerebral Ischemia Reperfusion model.
3, a kind of Rhesus Macacus middle cerebral artery ischemia model, it is to be made up by claim 1 or 2 described methods to obtain.
4, a kind of method of assessing the described Rhesus Macacus middle cerebral artery of claim 3 ischemia model is characterized in that: it is the assessment of adopting mainly the functional MRI method row model that relies on based on dispersion tensor imaging and blood oxygen level on the 3.0T magnetic resonance imager.
5, the application of the described Rhesus Macacus middle cerebral artery of claim 3 ischemia model in screening treatment cerebral infarction medicine.
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| CN101947223A (en) * | 2010-08-25 | 2011-01-19 | 新疆医科大学第一附属医院 | Composite anesthetic, preparation method thereof, and use thereof in myocardial infarction animal model |
| CN107374767A (en) * | 2017-08-22 | 2017-11-24 | 云南省第人民医院 | A kind of machin cerebral arterial thrombosis disease model and its method for building up |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101947223A (en) * | 2010-08-25 | 2011-01-19 | 新疆医科大学第一附属医院 | Composite anesthetic, preparation method thereof, and use thereof in myocardial infarction animal model |
| CN101947223B (en) * | 2010-08-25 | 2013-05-22 | 新疆医科大学第一附属医院 | Composite anesthetic, preparation method thereof, and use thereof in myocardial infarction animal model |
| CN107374767A (en) * | 2017-08-22 | 2017-11-24 | 云南省第人民医院 | A kind of machin cerebral arterial thrombosis disease model and its method for building up |
| CN109172032A (en) * | 2018-10-13 | 2019-01-11 | 刘劼 | A kind of microballoon pipe line pin device of MCAO models in rats |
| CN112998853A (en) * | 2021-02-25 | 2021-06-22 | 四川大学华西医院 | 2D modeling method, 3D modeling method and detection system for abdominal vascular dynamic angiography |
| CN112998853B (en) * | 2021-02-25 | 2023-05-23 | 四川大学华西医院 | Abdominal angiography 2D modeling method, 3D modeling method and detection system |
| CN115362987A (en) * | 2022-10-26 | 2022-11-22 | 首都医科大学宣武医院 | Preparation method and application of thrombus |
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