CN105203774B - Application of disheveled protein as stroke biomarker - Google Patents
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention belongs to the fields of medicine and biology, and relates to application of disheveled protein (DVL) as a cerebral apoplexy biomarker. In particular, the invention relates to the use of disheveled protein as a biomarker for stroke diagnosis, risk assessment and prognosis judgment. The invention also relates to application of the disheveled protein serving as a target spot of drug screening to screening drugs for treating cerebral apoplexy.
Description
Technical field
The invention belongs to medical science, biological field, and in particular to disheveled protein as apoplexy diagnose, risk assessment and
The purposes of the biomarker of Index for diagnosis.The invention further relates to disheveled protein is used to screen treatment as the target spot of drug screening
The purposes of the medicine of apoplexy.
Background technology
In the narrow sense, biomarker refer to detect blood or urine by chemistry or biological method predict physiology or
One class indicator of pathological state and disease risk degree.Biomarker is the effective tool in drug development process, and it is carried
The relevant information of pharmaceutical properties and disease process has been supplied, specific medication effect has been reflected.Diabetes and immune disease
All treated according to biomarker etc. various diseases.However, biomarker is still relative in cerebrovascular lacking.
And apoplexy is the third-largest high fatality rate in the whole world, high disability rate disease[1], cerebral infarction is the common type of apoplexy, sternly
Human health is threatened again[2].Therefore, the biomarker for exploring cerebral infarction has great significance.
Disheveled protein (disheveled, DVL) participates in intracellular signal transduction and right as the regulatory protein of Wnt paths
Downstream the transcription of the target factor, at aspects such as cell propagation, tumor formation, fetal development and Neural Differentiation important work is played
With[3,4].1 kind of signal transducer that DVL is made up of 670 amino acid residues, the wide expression in body tissue cell.
DVL mainly includes 3 conserved domains：DIX (Dis/Axin homologous domain), PDZ (PSD-95 and ZO-
1domain) and DEP, and between DIX and PDZ domains rich in serine/threonine area, under PDZ domains
The proline rich area of trip and the highly conserved C- end regions positioned at DEP domains downstream[5], different structure territory respectively with not
Same signal protein combines to play a part of to adjust cell.But at present still not using DVL as apoplexy biomarker
Report.
The content of the invention
The present inventor through groping and many experiments repeatedly, it was thus unexpectedly found that disheveled protein can be used as brain
The biomarker of apoplexy, for the judgement etc. of the diagnosis of apoplexy, the assessment of risk and prognosis, this completes this
It is bright.
A first aspect of the present invention is related to disheveled protein or its active fragment or detection disheveled protein or its active fragment
Purposes of the material in reagent preparation box, the test kit is used to diagnose apoplexy and/or the risk of apoplexy is suffered from assessment.
In one embodiment of the invention, wherein described test kit be according to the content of disheveled protein in sample or
The variation tendency of content suffers from the risk of apoplexy diagnosing apoplexy and/or assessment.
In one embodiment of the invention, wherein described sample is blood, serum, blood plasma and complete are selected from
Blood.
In one embodiment of the invention, described sample source is in mammal, such as people, mice, rat etc..
In embodiments of the invention, when the content of disheveled protein in sample is reduced or its content is less than normal value,
Show experimenter with apoplexy, or suffer from the risk height of apoplexy.
In one embodiment of the invention, described apoplexy is cerebral infarction.
A second aspect of the present invention is related to disheveled protein or its active fragment or detection disheveled protein or its active fragment
Purposes of the material in reagent preparation box, the test kit is used to assess the therapeutic effect of apoplexy or judges apoplexy
Prognosis.
In one embodiment of the invention, wherein described test kit be according to the content of disheveled protein in sample or
The variation tendency of content is assessing the therapeutic effect of apoplexy or judge the prognosis of apoplexy.
In one embodiment of the invention, wherein described sample is blood, serum, blood plasma and complete are selected from
Blood.
In one embodiment of the invention, described sample source is in mammal, such as people, mice, rat etc..
In one embodiment of the invention, when the content of disheveled protein in sample is raised, controlling for apoplexy is shown
Treat effectively or effect is preferable, or show that the prognosis of apoplexy is preferable.
In one embodiment of the invention, described apoplexy is cerebral infarction.
A third aspect of the present invention is related to disheveled protein or its active fragment or detection disheveled protein or its active fragment
Material be used for screening prevention or treat apoplexy medicine purposes or the purposes in reagent preparation box, wherein described
Test kit is used for screening prevention or treats the medicine of apoplexy.
In one embodiment of the invention, it passes through the content of disheveled protein or the change of content in detection sample and becomes
Gesture treats the medicine of apoplexy to screen.
In one embodiment of the invention, wherein described sample is blood, serum, blood plasma and complete are selected from
Blood.
In one embodiment of the invention, described sample source is in mammal, such as people, mice, rat etc..
In one embodiment of the invention, disheveled protein can be with the target spot for drug screening, for screening prevention
Or the medicine for the treatment of apoplexy.When the level of disheveled protein in sample after applying medicine is raised, show that the medicine can conduct
Prevention or the medicine for the treatment of apoplexy.
In one embodiment of the invention, described apoplexy is cerebral infarction.
A fourth aspect of the present invention relates to improve the material of disheveled protein content in blood to be used to prepare prevention or controls
Treat the purposes of the medicine of apoplexy.
In one embodiment of the invention, wherein the blood is selected from serum, blood plasma and whole blood.
In one embodiment of the invention, described sample source is in mammal, such as people, mice, rat etc..
In embodiments of the invention, it is possible to increase the material of disheveled protein content can be any material in blood,
Such as micromolecular compound, biomacromolecule such as protein (such as antibody, part), polypeptide or nucleic acid, the material can lead to
The content of disheveled protein during direct effect or indirect action with disheveled protein are crossed to improve blood.
In one embodiment of the invention, the material that can improve disheveled protein content in blood is Monot
Glycosides.
In one embodiment of the invention, described apoplexy is cerebral infarction.
In the present invention, the apoplexy is referred to because cerebral vessels rupture suddenly or because angiemphraxises cause blood circulation
Obstacle and cause one group of disease of brain tissue impairment.In embodiments of the invention, the apoplexy is cerebral infarction,
The cerebral infarction is referred to because brain blood circulation obstacle (is such as moved in internal carotid artery, external carotid artery, vertebral artery, brain
The circulatory disturbance that arteries and veins causes because of narrow or obturation), the focal brain tissue ischemia caused by ischemia, anoxia is downright bad or softening total
Claim, it is the main Types of apoplexy, accounts for the 60%～70% of apoplexy.
In the present invention, the disheveled protein derives from mammal, particularly people, and its sequence is, for example, AAH17225.1
It is shown.The active fragment of the disheveled protein is referred to can be in blood after the fragment with disheveled protein function, or degraded
The fragment for detecting, it can be a part for disheveled protein, or the aminoacid sequence of disheveled protein is through lacking, adding
Plus or the fragment that obtains after replacing；Such as active fragment is the piece of the part comprising disheveled protein and part or receptor binding
Section, or still retain the fragment of disheveled protein function through the disappearance of aminoacid, addition or after replacing.
In the present invention, the material of the detection disheveled protein or its active fragment is well known in the art, or its system
Preparation Method or preparation method are well known in the art, it is generally the case that it can be with disheveled protein or its active fragment specificity
With reference to by detecting that the material can detect disheveled protein or presence or the content of its active fragment, such as it can be the food in one's mouth
Naturally occurring part, receptor or the antibody that can be combined with disheveled protein or its active fragment in newborn animal body, or
Prepared by those skilled in the art can be with the antibody of disheveled protein specific binding, for example, monoclonal antibody or Anti-TNF-α
Body.In embodiments of the invention, the material of the detection disheveled protein or its active fragment is polyclonal antibody, its combination
Site is located between the 1-100 amino acids of people's Dvl albumen.
The beneficial effect of the invention
The present invention has been experimentally confirmed disheveled protein can be used as the biomarker of apoplexy, examining for apoplexy
The disconnected, assessment of risk and the judgement of prognosis；Meanwhile, disheveled protein is also used as the target spot of screening of medicaments, for screening
Prevention or the medicine for the treatment of apoplexy.
Description of the drawings
Fig. 1. the expression of different time points each group rat blood serum DVL albumen, n=4 after modeling；*p<0.05,*p<
0.01, compared with sham operated rats.
Fig. 2. the expression of the 7th day each group rat blood serum DVL albumen, n=4 after modeling；* *p<0.001, with sham-operation
Group is compared；###p<0.001, compared with model group.
12 points of point system results of Fig. 3 .Ludmila Belayev,* *p<0.001 compared with sham operated rats；###p<
0.001, compared with model group.
Fig. 4. morroniside to models of cerebral ischemia-reperfusion injury rat cerebral infarction volume MIR testing result,*p<0.001 with
Sham operated rats are compared；##p<0.01, compared with model group.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.It is unreceipted concrete in embodiment
Condition person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or the unreceipted production firm person of instrument, are
Can pass through city available from conventional products.
Embodiment 1DVL can be used as the biomarker of apoplexy
1 materials and methods
1.1 laboratory animal
SPF levels male Sprague-Dawley rat 20,7-8 week old, quality is 260-280g.Test purchased from Si Beifu
Animal Science and Technology Ltd., quality certification numbering：SCKK (capital) 2011-0004.Conventinal breeding, 24 ± 1 DEG C of ambient temperature, humidity 55
± 5%, water is can't help in preoperative 12h fasting.
1.2 medicine
Morroniside is voluntarily extracted from Fructus Corni by medicine research department of Xuan Wu hospital and prepared, efficient liquid phase chromatographic analysis purity
98.5%, the medicinal liquid of desired concn is dissolved into distilled water before use for experiment.
Chinese medicine Fructus Corni is the dry of Cornaceae plant Fructus Corni (Cornus officinalis Sieb.et Zucc.)
Dry mature fruit.Medicine research department of Xuan Wu hospital is further separated to its effective ingredient cornel iridoid glycoside, is obtained
Monomeric compound morroniside.Our early-stage Study finds that morroniside can reduce brain of rat with focal cerebral ischemia reperfusion infarction
Volume, strengthens cortex antioxidation, antiinflammatory and anti-apoptotic ability[6-8], and promote endogenic cell proliferation of nerve cord[9], improve god
Jing functions.Additionally, permeability of the morroniside in ischemia initial adjustment blood brain barrier, promotes blood brain barrier reconstruct, in the ischemia later stage
Strengthen endothelial progenitor cells propagation, promote angiogenesiss, finally improve microvascular function globality[10].The structure of morroniside is such as
Shown in lower：
1.3 reagents and instrument
Rabbit source property Dvl polyclonal antibodies (ab106844), mouse Tubulin monoclonal antibody (ab56676):The U.S.
Abcam companies；Horseradish peroxidase-labeled goat antirabbit two resists (ZB-2301), horseradish peroxidase-labeled mountain sheep anti mouse
Two anti-(ZB-2010)：Bioisystech Co., Ltd of Zhong Shan Golden Bridge of Beijing；RIPA lysates (P0013B):Green skies biology skill
Art institute；BCA methods protein quantification test kit (P1511):Beijing Puli's lema gene Technology Co., Ltd.；ECL Western
Blotting Kit(32109)：THERMO-PIERCE companies of the U.S..
Desk-top micro refrigerated centrifuger (Beckman Coulter companies, the U.S.)；(Bio-Rad is public for Powerpac Basic
Department, the U.S.)；Chemiluminescence gel imaging system (ProteinSimple companies, the U.S.)；All-wave length microplate reader (Thermo
Fisher companies, the U.S.).
1.4 experimental technique
1.4.1 it is prepared by Focal Cerebral Ischemia-Reperfusion in Rats model
With reference to Longa line brush[11]Prepare intraluminal middle cerebral artery occlusion in rats obstruction (middle cerebral artery
Occlusion, MCAO) model.Sprague-Dawley male rat operation consents fasting 12 hours.10% chloral hydrate 4ml/kg
Carry out intraperitoneal injection of anesthesia.Postanesthetic rat is lain on the back and is placed in surgical console, carried out being sterilized around cervical region with ANER DIAN.
In rat neck median incision about 2cm, muscle and fascia are separated, note avoiding thyroid.When trachea right side and breastbone
After trigonum exposure between musculus hyoideus and sternocleidomastoid, continue blunt separation and go out right carotid (Common
Carotid artery, CCA), internal carotid artery (Internal carotid artery, ICA) and external carotid artery (External
Carotid artery, ECA).The close crotches of ligation ECA and CCA proximal parts, close ICA, in CCA ophthalmology with bulldog clamp folder
The fine incision of a diagonal is cut, bolt line is carefully inserted, slowly touched bolt line and enter ICA, until producing sense, explanation are stopped
Bolt line passes through CCA crotches, by ICA enter cranium enter middle cerebral artery (middle cerebral artery,
MCA), anterior cerebral artery (anterior cerebral artery, ACA) initial part is reached, all blood supplies of MCA are blocked,
Cause middle cerebral artery occlusion.Bolt line is now no longer pushed, in case puncturing blood vessel causes subarachnoid hemorrhage.By bolt line with
CCA is ligatured in the lump, unclamps bulldog clamp, records the thromboembolism time started.Suture muscles, and a small amount of penicillin is applied at wound, with pre-
Anti- wound infection.After thromboembolism 30min, bolt line is slowly outwards extracted with ophthalmic tweezers, realize Reperfu- sion.Carry out after sham operated rats anesthesia
Identical operation technique, but not plug line.Notice that insulation, rat anesthesia are placed in temperature control blanket before not waking up after MCAO operations.
1.4.2 packet and administration
After rat is clear-headed, with reference to the point-scores of Zealonga 5 nervous function damage scoring is carried out[12], 1 point：Put forward rat-tail and leave ground
Face about 33cm, it is impossible to be fully extended offside forelimb；2 points：Put forward rat-tail and leave ground about 33cm, offside forelimb flexing；3 points：Will be big
Mus are placed in ground, to offside walking；4 points：Rat is placed in into ground, is turn-taked to offside, in the shape that knocks into the back；5 points：Damage serious, it is right
Side quadriplegia.1 point and 5 points of rat is rejected, the animal that nervous function damage scores and meets requirement of experiment is completed, according to commenting
Divide height, be randomly divided into sham operated rats, 1d groups after model, 3d groups after model, 7d groups after model, morroniside (270mg/kg) group,
4 per group.After modeling 3h, morroniside group continuous gavage administration 7d is administered once daily, and sham operated rats and model group give equivalent
Normal saline.
1.4.3 Western blotting analyze DVL protein expressions in serum
Before each group is drawn materials, by rat with 10% chloral hydrate 3.5ml/kg intraperitoneal anesthesias, from abdominal aortic blood 5-6mL,
4 DEG C of Refrigerator store 2h are placed in, 15min is centrifuged with 4 DEG C of 4000r/min, take supernatant, in -20 DEG C of preservations.Take the blood for having prepared
Clearly, after freeze thawing, 200 times of distilled water diluting is used afterwards 3 times with 0.22 μm of membrane filtration.Take the determination of serum albumen for having diluted dense
Degree, total protein and 5X sample-loading buffers 1:4 dilutions, 95 DEG C of degeneration 10min.
DVL adopts 10%SDS-PAGE separation gels, concentrates glue 5%SDS-PAGE.Electrophoresis：Concentrate glue voltage 60V,
40min, separation gel voltage 90V, 90min separates albumen.Transferring film is carried out after electrophoresis with 0.45 μm of NC film；Defat of the band 5%
2h is closed in milk powder；Rabbit polyclonal DVL antibody (1 is used respectively:1000), mouse monoclonal Tubulin antibody (1:2000) with 4
DEG C refrigerator overnight incubation.TBST buffer solutions 3 times, each 10min.Next with corresponding horseradish peroxidase (HRP)
With reference to two anti-incubation at room temperature 2h, TBST buffer solutions 3 times, each 10min；With ECL colour developing 1min, nitrite ion is filtered off, coagulated
Glue imager is made film.Recovery analysis is carried out to protein band with Quanity One softwares.
1.5 statistical analysis
Experimental data carries out statistical disposition using the statistics softwares of SPSS 13.0, is as a result represented with Mean ± SEM.Between group
Sample average compares with one factor analysis of variance (one-way analysis of variance, ANOVA), with p<0.05 represents
It is statistically significant.
2. result
Different time points DVL protein expression change after 2.1 Western blotting observation cerebral ischemic reperfusion in rats
As shown in figure 1,1d after modeling, model group serum DVL expressing quantities substantially reduce (P<0.05)；After modeling 3d and
7d model group serum DVL expressing quantities significantly reduce (P<0.01).
2.2 Western blotting observe impact of the morroniside to DVL expression after ischemic brain injury
As shown in Fig. 2 after modeling 7d, model group serum DVL albumen significantly reduces (P<0.001), compared with model group, not
Promise glycosides (270mg/kg) group DVL expressions significantly raise (P<0.001).
3. morroniside promotes neurological functional recovery to reduce infarct size
3.1 materials and methods
3.1.1 laboratory animal
SPF level male Sprague-Dawley rats, 7-8 week old, quality is 260-280g.Purchased from Beijing, dimension tonneau China is real
Test zoo technical company limited, quality certification numbering：SCKK (capital) 2007-0001.Conventinal breeding, 24 ± 1 DEG C of ambient temperature is wet
Degree 55 ± 5%, water is can't help in preoperative 12h fasting.
3.1.2 medicine
Morroniside is voluntarily extracted from Fructus Corni by medicine research department of Xuan Wu hospital and prepared, efficient liquid phase chromatographic analysis purity
98.5%, the medicinal liquid of desired concn is dissolved into distilled water before use for experiment.
3.1.3 reagent and instrument
10% chloral hydrate, Xuan Wu Preparation Room in Hospital.
Nylon embolus line (lot number：2632-100, d=0.26mm, Shadong Biological Technology Co., Ltd., Beijing)；Nuclear magnetic resonance, NMR is swept
Retouch instrument 3.0Tesla imaging systems (Philip).
3.2 experimental technique
3.2.1 it is prepared by Focal Cerebral Ischemia-Reperfusion in Rats model
With reference to Longa line brush[11]Prepare intraluminal middle cerebral artery occlusion in rats obstruction (middle cerebral artery
Occlusion, MCAO) model.Sprague-Dawley male rat operation consents fasting 12 hours.10% chloral hydrate 4ml/kg
Carry out intraperitoneal injection of anesthesia.Postanesthetic rat is lain on the back and is placed in surgical console, carried out being sterilized around cervical region with ANER DIAN.
In rat neck median incision about 2cm, muscle and fascia are separated, note avoiding thyroid.When trachea right side and breastbone
After trigonum exposure between musculus hyoideus and sternocleidomastoid, continue blunt separation and go out right carotid (Common
Carotid artery, CCA), internal carotid artery (Internal carotid artery, ICA) and external carotid artery (External
Carotid artery, ECA).The close crotches of ligation ECA and CCA proximal parts, close ICA, in CCA ophthalmology with bulldog clamp folder
The fine incision of a diagonal is cut, bolt line is carefully inserted, slowly touched bolt line and enter ICA, until producing sense, explanation are stopped
Bolt line passes through CCA crotches, by ICA enter cranium enter middle cerebral artery (middle cerebral artery,
MCA), anterior cerebral artery (anterior cerebral artery, ACA) initial part is reached, all blood supplies of MCA are blocked,
Cause middle cerebral artery occlusion.Bolt line is now no longer pushed, in case puncturing blood vessel causes subarachnoid hemorrhage.By bolt line with
CCA is ligatured in the lump, unclamps bulldog clamp, records the thromboembolism time started.Suture muscles, and a small amount of penicillin is applied at wound, with pre-
Anti- wound infection.After thromboembolism 30min, bolt line is slowly outwards extracted with ophthalmic tweezers, realize Reperfu- sion.Carry out after sham operated rats anesthesia
Identical operation technique, but not plug line.Notice that insulation, rat anesthesia are placed in temperature control blanket before not waking up after MCAO operations.
3.2.2 rat function assessment scores
After rat is clear-headed, with reference to Ludmila 12 points of point systems of Belayev (Ludmila Belayev ' s method
)[13,14], carry 0 point of tail hanging (2 points)：Without obvious neurological deficit；1 point：Flexing that infarction contralateral limbs are slight；2 points：Infarction
Contralateral limbs flexing is obvious.Limbs are placed, and are divided into the sub- test (front, side) of vision and sub- test (front, the side) (8 of tactile
Point) 0 point：Animal limb placing response is normal；1 point：Response delay but less than 2s；2 points：Response delay and>2s.Proprioceptive sensation is sub-
0 point of test (2 points)：It is more same than limbs with offside strong；1 point：With offside than limbs slight force；2 points：With offside than limbs without
Power.The animal that nervous function damage scores and meets requirement of experiment is completed, according to scoring height, sham operated rats, mould is randomly divided into
Type group, morroniside (270mg/kg) group, 14 per group.After postoperative 3h, morroniside group continuous gavage administration 7d is administered once daily,
Sham operated rats and model group give the normal saline of equivalent.
3.2.3 impact of the MRI detections morroniside to rat ischemia cerebral infarction volume
7 days after ischemia, each group animal carries out magnetic resonance examination, using Philips Gyroscan Intera 3.0T superconductions
Magnetic resonance device.After rat anesthesia, take supination position head part and be fixed on self-control poly (methyl methacrylate) plate, Mus head is put in C3 annular surface coils
Centre, the scope of examination includes routine T1WI, T2WI and DWI.MRI inspection parameters are that T1WI adopts SE sequences, T R500ms, T E
20ms.T2WI adopts T SE sequences, T R1624ms, TE 100ms, FOV 120mm, RFOV 75%.DWI scannings adopt EPI sequences
Row, T R 1000ms, TE 74ms, FOV120mm, RFOV 55%, NSA12, dispersion coefficient b values are respectively 0,1000s/mm2.
Using EPI sequences, T R248ms, TE 30ms, per layer scans 40 times PWI, FOV 120mm, RFOV 70%, NSA 2, angle of twist
40 degree, sweep time 47s.All of above sequence is Coronal and is imaged, thickness 2mm, and layer is away from 0.2mm.After completing scanning, measurement
The ADC values of region of interest, select the obvious 2 aspects measurement DWI abnormal signals area of ischemia and filling defect area to account for the full brain of same floor
The percentage ratio of area.
3.3 result
3.3.1 impact of the morroniside to Level In Rats With Focal Cerebral Ischemia neurological functional recovery
Continuous gavage is given after morroniside 7d, is measured 12 points of Ludmila Belayer and is scored, model group (5.36 ±
0.61) comparing with sham operated rats (0.00 ± 0.00) has significant difference (P<0.001, n=11), show that model group rats have
Significantly nervous function damage.Administration group (1.36 ± 0.49) compares with model group, there is significant difference (P<0.001, n=
14), show that morroniside administration group rat function has significant recovery (Fig. 3).
3.3.2 ischemical reperfusion injury 7d cerebral infarctions volume MIR testing result
MCAO ischemia-reperfusions carry out MRI detections after being administered 7 days.MRI is 3.0 Tesla imaging systems, coronal to brain
Face and sagittal plane T1 and T2 weighting picture and Diffusion weighted imaging are scanned.By contrast, t2 weighted image is finally selected to evaluate not
With the volume (Fig. 4) of matched group Cerebral Region, it is seen that sham operated rats T2W2 are showed as no abnormality seen.Model group occurs in T1W1 pictures
High RST, and clear-cut margin.Morroniside intervention group high earth core dam range shorter, and edge blurry.Statistical result showed (Fig. 4) is used
After medicine, ischemic region volume is compared compared with model group and significantly reduces (P<0.01), with significant difference.
4 conclusions
We have found that 1-7d serum DVL protein expressions are persistently reduced after infarction area after cerebral middle artery occlusion in rats at experimental result, illustrate big
Brain is in persistence faulted condition, and giving can significantly improve the expression of DVL while function of nervous system is improved after morroniside
Level.Above test result indicate that DVL can be used as apoplexy diagnosis and a kind of biomarker of prognosis instruction.
Although the specific embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.Root
According to disclosed all teachings, various modifications and replacement can be carried out to those details, these change in the guarantor of the present invention
Within the scope of shield.The four corner of the present invention is given by claims and its any equivalent.
List of references
[1]Navarro-Sobrino M,Rosell A,Hernández-Guillamon M,et al.A large
screening of angiogenesis biomarkers and their association with neurological
outcome after ischemic stroke[J].Atherosclerosis,2011,216(1):205-211.
[2]Lo EH,Dalkara T,Moskowitz MA.Mechanisms,challenges and
opportunities in stroke[J].Nature Rev Neurosci,2003,4(5):399-415.
[3] Ai Houxi, Li Lei, Xu Dongming, etc. morroniside is to Focal Cerebral Ischemia-Reperfusion in Rats cortex total antioxidation energy
Power affects [J]. China Rehabilitation theory and practice, 2009,15 (9)：833-834.
[4]Martin BL,Kimelman D.Canonical Wnt signaling dynamically controls
multiple stem cell fate decisions during vertebrate body formation[J].Dev
Cell,2012,22(1):223-232.
[5]Huang MY,Yen LC,Liu HC,et al.Significant overexpression of DVL1in
Taiwanese colorectal cancer patients with liver metastasis[J].Int J Mol Sci,
2013,14(10):20492-20507.
[6]Choi SH,Choi KM,Ahn HJ.Coexpression and protein-protein complexing
of DIX domains of human Dvl1and Axin1protein[J].BMB Reports,2010,43(9):609-
613.
[7] Ai Houxi, Wang Ying, Xu Dongming, etc. shadow of the morroniside to Focal Cerebral Ischemia-Reperfusion in Rats cortex IL-I β
Ring [J]. China Rehabilitation theory and practice, 2010,16 (10)：928-930.
[8] Wang Ying, high Dongming, Xu Dongming, etc. morroniside is activated to Focal Cerebral Ischemia-Reperfusion in Rats caspase-3
The impact [J] of degree. China Rehabilitation theory and practice, 2010,16 (9)：801-802.
[9]Sun FL,Wang W,Zuo W,et al.Promoting neurogenesis via Wnt/β-catenin
signaling pathway accounts for the neurorestorative effects of morroniside
against cerebral ischemia injury[J].Eur J Pharmacol.2014,738:214-221.
[10]Sun FL,Wang W,Cheng H,et al.Morroniside improves microvascular
functional integrity of the neurovascular unit after cerebral ischemia[J]
.PLoS One,2014,9(6):e101194.
[11]Cao G,Minami M,Pei W,et al.Intracellular bax translocation after
transient cerebral ischemia:Implications for a role of the mitochondrial
apoptotic signaling pathway in ischemic neuronal death[J].J Cereb Blood Flow
Metab,2001,21:321-333.
[12]Lo EH.A new penumbra:transitioning from injury into repair after
stroke[J].Nat Med,2008,14(5):497-500.
[13]Dieter-Chichung Lie.Wnt signalling regulates adult hippocampal
neurogenesis[J].Nature,2005,437(27):1370-1375.
[14]Maretto,S.et al.Mapping Wnt/β-catenin signalling during mouse
development and in colorectal tumors[J].Proc Natl Acad Sci USA,2003,100(6):
3299–3304。
Claims (9)
1. the material of disheveled protein or its active fragment or detection disheveled protein or its active fragment is in reagent preparation box
Purposes, the test kit is used for diagnosing ischemia apoplexy and/or the risk of cerebral infarction is suffered from assessment, also, described
Test kit be come diagnosing ischemia apoplexy and/or assessment according to the content of disheveled protein in sample or the variation tendency of content
The risk of cerebral infarction is suffered from, described sample is blood.
2. the purposes of claim 1, wherein described sample is selected from serum, blood plasma and whole blood.
3. the material of disheveled protein or its active fragment or detection disheveled protein or its active fragment is in reagent preparation box
Purposes, the test kit is used to assess the therapeutic effect of cerebral infarction or judge the prognosis of cerebral infarction, also,
Described test kit is assessing controlling for cerebral infarction according to the content of disheveled protein in sample or the variation tendency of content
Therapeutic effect judges the prognosis of cerebral infarction, and described sample is blood.
4. the purposes of claim 3, wherein described sample is selected from serum, blood plasma and whole blood.
5. the material of disheveled protein or its active fragment or detection disheveled protein or its active fragment is used for screening prevention or controls
The purposes or the purposes in reagent preparation box of the medicine of cerebral infarction are treated, wherein described test kit is used to screen pre-
Medicine that is anti-or treating cerebral infarction；Also, content or content that the test kit passes through disheveled protein in detection sample
Variation tendency screening the medicine for the treatment of cerebral infarction, described sample is blood.
6. the purposes of claim 5, wherein described sample is selected from serum, blood plasma and whole blood.
7. the material of disheveled protein content in blood can be improved to be used to prepare the medicine of prevention or treatment cerebral infarction
Purposes.
8. the purposes of claim 7, wherein the blood is selected from serum, blood plasma and whole blood.
9. the purposes of claim 7, wherein the material that can improve disheveled protein content in blood is morroniside.
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