CN103784974A - Application of interferon regulatory factor 8 (IRF8) in cerebral apoplexy disease - Google Patents

Abstract

The invention discloses function and application of an IRF8 gene in a cerebral apoplexy disease, belonging to the field of the function and the application of a gene. According to the invention, IRF8 gene knockout mice and neuron-specific IRF8 transgenic mice are taken as experimental subjects, a brain middle artery ischemia reperfusion model is adopted, and the results show that the cerebral infarction volume of the IRF8 gene knockout mice is obviously increased and the neurological function is obviously worsened as well in comparison with that of the wild C57 mice, the infarction volume of the IRF8 transgenic mice is obviously reduced, and the neurological function obviously gets better. The invention discloses the function of the IRF8 gene in the cerebral apoplexy disease, which mainly means that the IRF8 gene has an effect of protecting the function of a nervous system, especially an effect that the IRF8 gene can protect the cerebral apoplexy disease. According to the abovementioned function of the IRF8, the invention provides the application of the IRF8 in preparation of a drug for treating the cerebral apoplexy disease.

Description

The application of interferon regulatory factor 8 (IRF8) in apoplexy disease

Technical field

The invention belongs to function and the application of gene, particularly a kind of interferon regulatory factor 8(interferon regulatory factor 8, IRF8) application in apoplexy disease.

Background technology

Cerebral infarction is the main lethal disease disabling in the whole world at present, (tissue-the typeplasminogenactivator of tissue plasminogen activator at present, tPA) fibrinolytic is still the primary treatment method for the treatment of ischemic cerebrovascular, but the ischemia-reperfusion of following can further increase the weight of ischemia neural cell injury.Research shows, neuro-protective strategy can improve brain function after cerebral ischemia in the long period, reduces neuronal cell loss.Apoptosis is one of fundamental mechanism of cell death in cerebrum ischemia/refilling process, but its regulatory mechanism is illustrated not yet completely.Therefore, when research cerebrum ischemia/pour into again, the molecular mechanism of neuronal cell apoptosis existence, provides new therapeutic strategy and method by contributing to for neuro-protective.

Cerebrum ischemia reperfusion injury can cause multiple difference but overlapped signal path, modulating apoptosis in platelets or death.When neuron focal cerebral ischemia, infarction core space overwhelming majority cells all necrose (necrosis), and feature is that energy supply reduces sharply and causes cellular edema, organelle to break and the irreversible death of cell.The low perfusion cerebral tissue of infarction core periphery is called " ischemia half blanking bar ", and because it still has metabolic activity, if cerebral blood perfusion improves, this region cytoactive still can be recovered.Therefore save ischemia half blanking bar for apoplexy after treatment there is important value.Although 1996 Nian Qi tissue plasminogen activators (tPA) ratify for cerebral infarction treatment, it remains the thrombolytic drug that the unique audit of Bureau of Drugs Supervision of the U.S. (FDA) is passed through up to now.Increase because extend in time bleeding risk, the therapeutic time window of tPA is only 4.5 hours; Consider the difficult discriminating of iconography in early days of ischemic and hemorrhagic apoplexy, further incured loss through delay patient and accepted the chance that tPA treats.Only be less than at present 5% ischemic cerebral stroke patients use tPA thromboembolism treatment.In addition, if the long-time severe ischemic anoxia of cerebral tissue is found in research, still can cause irreversible damage to cerebral tissue even recover cerebral blood flow in the later stage, therefore current still in the urgent need to studying the therapeutic strategy of the pathophysiology event for hypoxic-ischemic and (or) due to pouring into again.Since the nineties in 20th century, the therapeutic strategy of research neuroprotective and cerebral tissue is the focus of Treatment of Cerebral Stroke always, and these strategies not only can extend the therapeutic time window of tPA, also can alleviate the brain tissue impairment of ischemia-reperfusion induction.Multiple nerve protection medicine has been obtained stem-winding result in zoopery; but enter after apoplexy 3 phase clinical experiment; overwhelming majority medicine does not all attain the results expected; one of its primary failed reason is that most of known Neuroprotective Mechanisms acted on after apoplexy in 4-6 hour; and in clinical practice, be difficult to so of short duration be in time window, to implement treatment, after therefore further illustrating apoplexy and occurring, in longer a period of time, promote or the molecular mechanism of protection brain tissue impairment significant for the effective Treatment of Stroke target spot of research or strategy.

Interferon regulatory factor (interferon regulatory factor, IRF) family has had now found that 10 members, and it consists of IRF1～IRF10.Existing research prompting, IRF family member has participated in biological process widely, relates generally to the natural immunity and the acquired immune response, and regulating cell growth and existence, apoptosis and propagation participate in hemopoietic, antitumor formation etc.IRF8 be the common recognition of interferon not in conjunction with albumen (interferon consensus sequence-binding protein, ICSBP), be a member of IRF family, as a transcription factor, thereby can play a role by transcriptional control DNA.IRF8 is found to be in myelocyte and lymphocyte first, there are some researches show that IRF8 plays central role in immunomodulating and myelocyte differentiation.Multinomial research shows, IRF8 may participate in multiple sclerosis, central nervous system's inflammatory demyelination, the diseases such as peripheral nerve injury.

Summary of the invention

For solving defect and the deficiency of above-mentioned prior art, primary and foremost purpose of the present invention is to provide the application of a kind of IRF8 in preparation treatment nervous system disease medicine.

Another object of the present invention is to provide the application of a kind of IRF8 in preparation treatment apoplexy disease medicament.

Object of the present invention is achieved through the following technical solutions:

The present invention is take IRF8 knock out mice and Neuron-specific IRF8 transgenic mice as experimental subject, by Cell transplantation model, result shows to contrast with wild type C57 mice, IRF8 knock out mice head infarction obviously increases the weight of, function of nervous system also obviously worsens, the Infarction volume of Neuron-specific IRF8 transgenic mice is suppressed, and function of nervous system also takes a turn for the better.This prompting IRF8 gene has the effect of neuroprotective systemic-function, and the nerve injury that can protect cerebral ischemia to cause, for new drug and the New Policy of research control cerebral ischemia provide theoretical foundation and Clinical Basis.

The function of IRF8 gene in apoplexy disease, is mainly reflected in IRF8 gene and has the effect of neuroprotective systemic-function, and particularly IRF8 gene can be protected the effect of Imaging in Patients with Cerebral Ischemia Disease.

For the above-mentioned functions of IRF8 gene, provide IRF8 to apply in the medicine of preparation treatment nervous system disease.

A medicine for the treatment of nervous system disease, comprises IRF8.

For the above-mentioned functions of IRF8 gene, provide IRF8 to apply in preparation treatment apoplexy disease medicament.

A medicine for the treatment of apoplexy disease, comprises IRF8.

Achievement in research of the present invention shows, in the damage that IRF8-KO mice causes at Cell transplantation, mice Infarction volume obviously increases the weight of, and function of nervous system obviously worsens, and neuronal apoptosis also obviously increases.Prove that IRF8 gene has important protective effect in apoplexy disease model.

The present invention has following advantage and effect with respect to prior art:

1. the present invention finds the new function of IRF8 gene, and IRF8 gene can be protected the effect of apoplexy disease.

2. the effect of IRF8 in protection apoplexy disease, for the preparation for the treatment of apoplexy disease medicament.

Accompanying drawing explanation

Fig. 1 is structure and the qualification result figure of nerve-specific IRF8 transgenic mice

A is the design of graphics of nerve-specific IRF8 transgenic mice;

B is the qualification result figure of nerve-specific IRF8 transgenic mice;

Fig. 2 is the TTC coloration result figure of WT and IRF8-KO mice.

A is TTC coloration result figure;

B is cerebral infarction volume statistics block diagram;

C is function of nervous system's scoring statistics block diagram;

Fig. 3 is the TTC coloration result figure of IRF8-TG and NTG mice.

A is TTC coloration result figure;

B is cerebral infarction volume statistics block diagram;

C is function of nervous system's scoring statistics block diagram;

Fig. 4 is the cerebral tissue infarction surrounding zone neuronal cell apoptosis situation measurement result figure of WT and IRF8-KO mice.

Figure A is Fluoro Jade B detection display figure and statistical result figure;

Figure B is TUNEL apoptosis figure and statistical result figure;

Fig. 5 is the cerebral tissue infarction surrounding zone neuronal cell apoptosis situation measurement result figure of IRF8-TG and NTG mice.

Figure A is Fluoro Jade B detection display figure and statistical result figure;

Figure B is TUNEL apoptosis figure and statistical result figure;

The specific embodiment

Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited to this.

Animal for research and raising

Laboratory animal: select age in 11-12 week, body weight at 25-30g; background is the wild-type mice (WT of male C57BL/6 strain; purchased from Fukang bio tech ltd of Beijing China), IRF8 knock out mice (IRF8-KO; C57BL/6J background; purchased from EMMA; article No. EM:02414), non-transgenic mice (NTG, Neuron-specific Cre transgenic mice (CaMKII α-Cre; Purchased from Jackson Laboratory, Stock No. 005359)) and nerve-specific IRF8 transgenic mice (IRF8-TG, obtained by IRF8-flox transgenic mice and the hybridization of CaMKII α-Cre mice, IRF8-flox transgenic mice is built by this laboratory oneself, and the building process of IRF8-flox transgenic mice as described hereinafter).

Feeding environment: all experiment mices are all raised in the SPF of angiocardiopathy institute of Wuhan University level Experimental Animal Center.Mice special feed is provided by Chinese military medicine academy of science animal center.Raising condition: room temperature is between 22-24 ℃, and humidity is between 40-70%, and it is 12h that light and shade replaces lighting hours, freely drinks water and ingests.

The structure of [embodiment 1] nerve-specific IRF8 transgenic mice

IRF8-flox transgenic mice builds information:

Transgene carrier builds information: with forward primer, i.e. 5 '-CCAGATTACGCTGATTGTGACCGGAACGGCGGGCG-3 ' (SEQ ID NO. 1); Downstream primer, 5 '-AGGGAAGATCTTGATTTAGACGGTGATCTGTTGAT-3 ' (SEQ ID NO. 2), amplification mice IRF8 full-length gene (NCBI, Gene ID:15900, NM_008320.3), cDNA is inserted to pCAG-CAT-LacZ carrier, β-actin gene (CAG that this carrier comprises a cmv enhancer and a chicken, chicken β-actin gene) promoter, and be connected to chloramphenicol acetyl transferasegene (CAT, chloramphenicol acetyltransferase), loxP site is positioned at CAT both sides.The expression of neurocyte IRF8 obtains (Figure 1A) by CAG promoters driven.IRF8-floxed mouse: the pCAG-IRF8-CAT-LacZ carrier of structure, by the microinjection embryo's (C57BL/6J background) that is configured to be fertilized, is obtained to IRF8-floxed transgenic mice.Neuronal specificity IRF8 transgenic mice obtains by IRF8-flox mice and CaMKII α-Cre mice outbreeding.

Transgenic mice is got genomic DNA by cutting tail, use PCR to identify, PCR identifies that primer information is: detect forward primer 5 '-CCAGATTACGCTGATTGTGACCGGAACGGCGGGCG-3 ' (SEQ ID NO. 3), detect reverse primer 5 '-AGGGAAGATCTTGATTTAGACGGTGATCTGTTGAT-3 ' (SEQ ID NO. 4).Expression by IRF8 albumen in the different transgenic mouse heads of western blotting (Western Blot) experimental identification: extract different transgenic mouse cerebral tissue albumen, by polyacrylamide gel electrophoresis (SDS-PAGE), checking IRF8 crosses expression (Figure 1B)

We have built a few strain nerve-specific IRF8 transgenic mices (IRF8-TG).In order to reflect the change of IRF8 under pathological and physiological condition, we have selected IRF8-TG5 mice, Western Blot and quantitative analysis demonstration, and in its cerebral tissue, IRF8 expression is about 10.5 times of normal structures.

[embodiment 2] mouse brain Infarction Model (I/R) obtains

1. laboratory animal grouping: male C57BL/6 strain wild-type mice, IRF8 knock out mice and head specificity IRF8 transgenic mice and non-transgenic mice, set up Cerebral Infarction Model (I/R) by Cell transplantation.Be divided at random 8 groups, every group of 10 mices: C57BL/6 strain wild-type mice sham operated rats (WT SHAM) and I/R art group (WT I/R), IRF8 knock out mice sham operated rats (KO SHAM) and I/R art group (KO I/R), non-transgenic mice sham operated rats (NTG SHAM) and I/R art group (NTG I/R), neuronal specificity IRF8 transgenic mice sham operated rats (TG SHAM) and I/R art group (TG I/R).

2. line bolt method cerebral infarction I/R operation adopts mouse brain medium-sized artery ischemia-reperfusion (middle cerebral artery Ischemia Reperfusion) model manipulation flow process:

(1) capture mice, use 3% isoflurane anesthesia mice, 8% sodium sulfide is sloughed the Mus hair of cervical region, and calvarium Mus hair is cut rapidly with operating scissors, 3% povidone iodine sterilization cervical region and calvarium skin 2 times, the de-iodine of 75% ethanol 1 time;

(2) at the calvarium position of mice cross sections, expose skull, peel off gently the connective tissue of skull surface with tweezers.The fibre-optical probe of laser Doppler flowmetry is fixed on to bregma rear 2mm, the position of left side 5mm with biogum;

(3) mice is lain on the back fixing, neck median line otch, along sternocleidomastoid inner edge separating muscle and fascia, separates left carotid (CCA), external carotid artery (ECA) and internal carotid artery (ICA).Prick with 8-0 toe-in at ECA distal end, ECA proximal part place hanging wire is for subsequent use.Press from both sides temporary transient folder with arteriole and close ICA, CCA; Then in the middle of the ligation of ECA distal end and proximal part hanging wire, cut an osculum, line bolt is sent to CCA by clip, and the hanging wire of ECA proximal part is made a call to a slip-knot at clip, elasticity can free in and out with line bolt but the sense that slightly rubs is advisable, loose ICA bulldog clamp again, line bolt is sent into ICA, start to calculate distance from vascular bifurcation, stop to the decline power that is hampered of blood flow in about 9-11mm when insertion depth.At this moment will fasten gently and fasten line around ECA proximal part place slip-knot, whole process must maintain the anus temperature of mice at 37 ± 0.5 ℃;

(4) enter decline power stopping time that is hampered of cerebrovascular to blood flow from line bolt and start timing, after 45min, first unclamp ECA proximal part place slip-knot, line bolt is extracted, and ECA proximal part place slip-knot is tightened, unclamp rapidly CCA place bulldog clamp, and by the ligation of ECA proximal part (Sham group enter from line bolt cerebrovascular to blood flow decline take out Outlet bolt while being hampered power).Note observing restoration of blood flow situation, select blood flow to decline more than 75%, the mice that restoration of blood flow reaches more than 70% is included experiment in;

(5) sew up mice cervical region and skin of head, and with the povidone iodine wound of sterilizing.After operation finishes, mice is placed in incubator, case temperature maintains 28 ℃, and feedwater and feedstuff are to drawing materials.

[embodiment 3] Cerebral Infarction Model (I/R) mouse brain Infarction volume is measured

The evaluation index of the cerebral ischemia/reperfusion injury order of severity mainly comprises the scoring of infarction of brain volume and function of nervous system, these indexs all with ischemia/reperfusion injury order of severity positive correlation.

(1) 24h after operation respectively, carries out function of nervous system and shape for learning scoring before 72h draws materials;

Based on Berderson function of nervous system scoring improve one's methods (9 points of systems):

0 point: the symptom that impassivity is impaired;

1 point: while carrying tail, offside forelimb is curled, or can not arrive Ipsilateral forelimb completely;

2 points: while carrying tail, in offside shoulder, receive;

3 points: horizontal sliding: while promotion to offside, resistance declines;

4 points: can be spontaneous to all directions motions, but only turn to offside in the time of de-tail;

5 points: when autonomic movement, turn-take or only to turning;

6 points: without autonomic movement, only motion in the time stimulating;

7 points: without autonomic movement, when stimulation also without motion;

8 points: the death relevant with cerebral ischemia.

(2) capture mice, lumbar injection 3% pentobarbital sodium anesthetized mice, cuts off mice thoracic cavity, breaks heart blood-letting;

(3) skin of neck after volume fraction 75% alcohol disinfecting, cut off rear skin of neck, expose head and cervical region, cut off neck marrow from cervical vertebra, rear musculi colli is removed in separation, and eye scissors is longitudinally cut off the outer skull of brain stem cerebellum, with stricture of vagina tooth pincers strip off skull, separate brain surface's cerebral dura mater, avoid cerebral dura mater to scratch cerebral tissue.While getting brain, from oblongata, careful separation basis cranii tissue, avoids damaging brain;

(4) cerebral tissue taking off is put into the culture dish rinse that PBS is housed, blotted PBS with gauze, cerebral tissue is put into 1mm mouse brain mould, be placed in-20 ℃ of refrigerators frozen (being no more than 4h);

(5) cerebral tissue 2,3, 5-Triphenyltertrazoliumchloride (2,3,5-Triphenyltetrazolium chloricej, TTC) dyeing: take out cerebral tissue from-20 ℃ of refrigerators, be cut into immediately 1mm slab, comprise that bregma front cuts 4, rear is cut 3, cuts altogether 7.Section is placed in immediately to the serum bottle of 10mL2% TTC solution, 37 ℃ of constant-temperature incubation 10min.Frequently stir section, make even tissue dyeing.After normal cerebral tissue's dyeing, be cerise, and infarcted region is pale asphyxia;

(6) cerebral tissue is fixed: the cerebral tissue in beaker and solution are together proceeded in the cup of carrying out labelling, discard TTC solution, with the fixing brain tissue slice of 10% neutral formalin solution, take pictures and use IPP software analysis after 24h;

(7) cerebral infarction volume calculates: Infarction volume %=(not Infarction volume of offside cerebral hemisphere volume-infarction side)/(offside cerebral hemisphere volume × 2) × 100%;

Total Infarction volume is 7 brain sheet result data sums separately.

As shown in Figure 2, pour into IRF8-KO mice Infarction volume after 24 hours through I/R ischemia 45min increases compared with wild-type mice TTC coloration result again; And this deterioration acts on I/R and still continues for postoperative 72 hours, and function of nervous system's scoring all increases the weight of at I/R for postoperative 24 hours, 72 hours.

As shown in Figure 3, pouring into IRF8-TG mice Infarction volume after 24 hours through I/R ischemia 45min obviously alleviates compared with wild-type mice again; And this protective effect still continues at I/R for postoperative 72 hours, and function of nervous system's scoring all alleviates at I/R for postoperative 24 hours and 72 hours.

Embodiment 4. cerebral tissue infarction surrounding zone neuronal cell apoptosis situations are measured

1. cerebral tissue frozen section preparation

1) experiment mice is pressed the anesthesia of 50mg/kg dosage lumbar injection pentobarbital sodium;

2) open breast and expose heart, puncture as left ventricle with injection needle, cut off right atrium simultaneously;

3) use 0.1mol/l PBS(pH7.4) after 100mmHg pressure perfusion loses color to liver, with 4% paraformaldehyde perfusion 15min;

4) open cranium and take out rapidly mouse brain, after room temperature 4% paraformaldehyde, fix 6-8h;

5) olfactory bulb and the cerebellum of excision cerebral tissue, then prolong median line brain is divided into first latter two part, fixes 15min again with previous fixative;

6) be immersed in subsequently containing in the phosphate buffer of 30% sucrose, 4 ℃ of refrigerators sink to the bottom and spend the night;

7) after 30% sucrose mixes by 1:1 with OCT, in right amount in embedding frame, the tissue of back is taken out, suck liquid on gauze after, in this embedding frame, soak a little while, be transferred to again and first added in another embedding frame of 2 OCT, the position of adjusting tissue, makes it just in time be positioned at the center of embedding frame;

8) will contain the embedding frame of tissue, and move in dry ice, and make it in horizontal position as far as possible, and slightly, after a little while, continue to add OCT, certain height is organized in submergence, after OCT solidifies, is stored in the refrigerator of-80 ℃;

9) cut the frozen section of 5 μ m by the standardization program of freezing microtome for subsequent use.

2.TUNEL test kit staining examine apoptosis.

With TUNEL test kit staining examine apoptosis.(TUNEL test kit: ApopTag Plus In Situ Apoptosis Fluorescein Detection Kit (S7111, Chemicon)):

1) ice is cut to tissue slice and is placed in the paraformaldehyde of (pH 7.4) 1%, the fixing hydrolysis of room temperature 10 minutes;

2) PBS washes twice, each 5 min;

3) be placed in the ethanol of pre-cooling: acetic acid (2:1) solution ,-20 ℃ are soaked 5 minutes, remove unnecessary liquid, but note dry;

4) PBS washes twice, each 5 min;

5) filter paper carefully sucks unnecessary liquid, presses immediately 75 μ L/5 cm in section ²directly add level pad, incubated at room 1-5 min;

6) filter paper carefully sucks unnecessary liquid, presses immediately 55 μ l/5 cm in section ²directly add TdT enzyme reaction solution, be placed in lucifuge moisture preservation box effect 1 h(negative control and add the not reactant liquor containing TdT enzyme);

7) section is placed in to termination/lavation buffer solution, shakes gently 15 sec, incubated at room 10 min; Now prepare appropriate anti digoxin antibody, be preheated to room temperature, note lucifuge;

8) PBS washes three times, each 1 min;

9) filter paper carefully sucks unnecessary liquid, directly in section, presses 65 μ L/5 cm ²add anti digoxin antibody, under room temperature, in the wet box of lucifuge insulation, act on 1 h;

10) PBS washes four times, each 2 min;

11) SlowFade Gold antifade reagent with DAPI(Invitrogen, S36939) mounting;

12) fluorescence Microscopic observation, takes pictures.Preserve 4 ℃ of preservations in dark wet box if need.At fluorescence microscopy Microscopic observation, take pictures, counting Apoptotic neuron cell.(if needing to preserve 4 ℃ of preservations in dark wet box)

3. FJB(Fluoro Jade B) dyeing

1) ice is cut to tissue slice dries 1 hour in baking oven;

2) 1% NaOH+80% dehydrated alcohol 5min;

3) 70% dehydrated alcohol 2min;

4）dd H ₂O 2min；

5) Flouro jade B diluent (AG310, Millipore, Billerica, MA), room temperature lucifuge 20min;

6) dd H ₂o 1min washes 3 times;

7) in baking oven, dry sheet 5-10min;

8) dimethylbenzene is processed 2-3min;

9) mounting, takes pictures.

Cerebral tissue infarction surrounding zone neuronal cell apoptosis situation measurement result is shown in Fig. 4, Fig. 5.Fig. 4 is postoperative 24 hours cerebral tissue infarction surrounding zone neuronal cell apoptosis situations of IRF8-KO mice and wild-type mice I/R.Fluoro Jade B dyeing (A) and TUNEL dyeing (B) detect apoptosis, and result shows that IRF8-KO Mouse Neuron apoptosis rate all increases than wild-type mice, dead relevant while further pointing out IRF8 to neuronal cell ischemia/reperfusion.Same Fig. 5 is postoperative 24 hours cerebral tissue infarction surrounding zone neuronal cell apoptosis situations of IRF8-TG mice and NTG mice I/R, and Fluoro Jade B dyeing (A) and TUNEL dyeing (B) result show that IRF8-TG Mouse Neuron apoptosis rate all reduces than NTG mice.These results show, promote IRF8 to express and can improve cerebral tissue ischemia/reperfusion injury, and may be closely related with neuronal cell apoptosis.

Our achievement in research shows, in the damage that IRF8 KO mice causes at Cell transplantation, after we find that IRF8 knocks out, mice Infarction volume significantly increases, and function of nervous system obviously worsens, and neuronal apoptosis is showed increased also.Prove that IRF8 gene has important protective effect in apoplexy disease model.

Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.

SEQUENCE LISTING

<110> Wuhan University

<120> interferon regulatory factor 8(IRF8) application in apoplexy disease

<130> 1

<160> 4

<170> PatentIn version 3.3

<210> 1

<211> 35

<212> DNA

<213> Artificial

<223> IRF8 forward primer

<400> 1

ccagattacg ctgattgtga ccggaacggc gggcg 35

<210> 2

<211> 35

<212> DNA

<213> Artificial

<223> IRF8 downstream primer

<400> 2

agggaagatc ttgatttaga cggtgatctg ttgat 35

<210> 3

<211> 35

<212> DNA

<213> Artificial

<223> detects forward primer

<400> 3

ccagattacg ctgattgtga ccggaacggc gggcg 35

<210> 4

<211> 35

<212> DNA

<213> Artificial

<223> detects reverse primer

<400> 4

agggaagatc ttgatttaga cggtgatctg ttgat 35

Claims (4)

1. The application of 1.IRF8 in preparation treatment nervous system disease medicine.
2. 2. a medicine for the treatment of nervous system disease, is characterized in that: comprise IRF8.
3. The application of 3.IRF8 in preparation treatment apoplexy disease medicament.
4. 4. a medicine for the treatment of apoplexy disease, is characterized in that: comprise IRF8.

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