CN1150944C - Application of alkaline fibroblast growth factor in preparing medicine to prevent and cure cerebral injury - Google Patents
Application of alkaline fibroblast growth factor in preparing medicine to prevent and cure cerebral injuryInfo
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- CN1150944C CN1150944C CNB011147563A CN01114756A CN1150944C CN 1150944 C CN1150944 C CN 1150944C CN B011147563 A CNB011147563 A CN B011147563A CN 01114756 A CN01114756 A CN 01114756A CN 1150944 C CN1150944 C CN 1150944C
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Abstract
The present invention discloses the application of a basic fibroblast growth factor to preparing a medicine for preventing and treating cerebral injuries. The a basic fibroblast growth factor delays neuron death and promotes nerve fiber regeneration through protecting the neurons against injuries, and the a basic fibroblast growth factor used as a neuron protection medicament for preventing and treating cerebral injuries is applied for the first time.
Description
The present invention relates to the application of application, the especially basic fibroblast growth factor of a kind of somatomedin in preparation prevention, treatment brain injury medicine in preparation prevention, treatment brain injury medicine.
Basic fibroblast growth factor (basic fibroblast growth factor; bFGF) be a kind of widely distributed in vivo somatomedin; have the cell differentiation of promotion hypertrophy, angiogenesis, wound healing and vasorelaxation action, and damaged cell is had direct protective action.
The object of the present invention is to provide the application of a kind of basic fibroblast growth factor in preparation prevention, treatment brain injury medicine.
The application of basic fibroblast growth factor of the present invention in preparation prevention, treatment brain injury medicine, its midbrain injury is the damage that cerebral trauma, apoplexy, nerve regression disease or brain operation are brought, and apoplexy is cerebral ischemia or cerebral hemorrhage, and nerve regression disease is senile dementia or Parkinson's disease.
The present invention compared with prior art has the following advantages:
The present invention exempts from infringement by neuroprotective unit, delays neuronal death, promotes the mechanism of nerve fiber regeneration, is first as nerve protection medicine prevention, treatment brain injury.
The following examples will help those of ordinary skill in the art further to understand the present invention, but not limit the present invention in any form.
Embodiment 1
The application of basic fibroblast growth factor in preparation prevention, treatment focal cerebral ischemia medicine
1, material and method
1.1 70 of the normal adult SD rats that Zhongshan Medical Univ.'s animal center provides are selected in laboratory animal and grouping for use, body weight 300～400g, male and female are regardless of, be divided into 7 groups at random: cerebral ischemia perfusion or give the basic, normal, high dosage of bFGF, normal saline each group and sham operated rats and normal control group simultaneously again after 3 hours, 10 every group.
1.2 reaching perfusion modelling again, rat cerebral ischemia adopt Nagasawa ' s method to be improved.Separate, expose left carotid, ligation it, and separation ligation left side external carotid artery root, separate the left side internal carotid artery, separate the tie wings arteria palatina, about 2mm cuts an osculum in the place in left carotid apart from end, one sub-thread nylon filament is inserted common carotid artery, enter internal carotid artery through furcation, continuation pushes into cranial cavity gently along internal carotid artery, and insertion depth is about 22～25.0mm altogether, withdraws from filament perfusion more at once when ischemia reaches 3 hours, promptly at perfusion while femoral vein instillation bFGF and normal saline again, bFGF is low for bFGF group and normal saline group, in, high dose is respectively 5,50,250ug/Kg.
The killing live rat 1.3 TTC dyes takes out full brain fast, goes olfactory bulb and cerebellum, the crown section of whole brain tissue is divided into 6 parts (every agreement that contracts a film or TV play to an actor or actress 2mm), the brain sheet is immersed in the 2%TTC solution (normal saline preparation), hatched 30 minutes for 37 ℃, move in the formalin fixed liquid.Then the brain sheet is taken pictures, treat image analysis.
1.4 image analysis adopts the Tiger image analysis system to calculate brain sheet area and brain sheet ischemic region area, according to brain sheet THICKNESS CALCULATION ischemia percent by volume in brain sheet photo input computer.
1.5 statistical procedures adopts the t inspection statistics respectively to organize data.Respectively organize the ischemia percent by volume.
2, result
The image analysis system result shows that the cerebral infarct volume percentage ratio (%) of ischemic reperfusion group, ischemic reperfusion+normal saline group, the basic, normal, high dosage group of ischemic reperfusion+bFGF is respectively 25.66 ± 1.89,23.23 ± 2.64,14.69 ± 2.43,10.98 ± 3.10 and 13.57 ± 3.43.Statistical result shows the high, medium and low dosage group of bFGF and ischemic reperfusion+normal saline group relatively, and all difference is remarkable, and dosage group and low dose group compare among the bFGF, significant difference; Relatively, difference does not have significance (table 1) between bFGF height, the middle dosage group, between the high and low dose group.
Cerebral infarct volume percentage ratio between each group of table 1 (%, x+s) relatively
Group n infarct volume percentage ratio (%)
Cerebral ischemia reperfusion stream organizes 5 25.66 ± 1.89
Ischemic reperfusion+normal saline group 5 23.23 ± 2.64
BFGF low dose group 8 14.69 ± 2.43
Dosage group 5 10.98 ± 3.10 among the bFGF
BFGF high dose group 6 13.57 ± 3.43
The reason that bFGF reduces cerebral infarct volume: 1. bFGF is to neuronic direct protective action.Blazoning the FGF receptor family on neurocyte and the glial cell film, when brain injury took place, bFGF and expression of receptor thereof strengthened in the cerebral tissue; exogenous bFGF by with neuron membrane on receptors bind; the enabling signal pathway, neuroprotective unit exempts from destruction, and stimulating neuronal germinates.BFGF has significant protective effect to external death because of inductive Hippocampus of ischemia and cortical neuron in addition, and bFGF can suppress a kind of nmda receptor correlative protein expression, and suppresses Ca in the born of the same parents
2+Overload and promotion Radical Metabolism, so bFGF keeps Ca in the born of the same parents by antagonism irritability toxic action
2+Stablize and the promotion Radical Metabolism, the neuroprotective cell exempts from destruction.2. bFGF is to the influence of cerebral blood flow.BFGF has the effect of the little blood vessel of expansion pia mater encephali, thus the cerebral blood flow increasing amount, the neurocyte in the protection cerebral ischemia.In addition, bFGF has strong angiogenic growth and newborn effect.3. the high, medium and low dosage group of bFGF dose-effect relationship analysis bFGF cerebral infarct volume percentage number average significantly reduces than ischemic reperfusion+normal saline group, the bFGF that high, medium and low dosage is described all can stop the expansion of cerebral infarction scope, and middle dosage group obviously reduces than the cerebral infarct volume percent of low dose group, and no significant difference between senior middle school's dosage group, between the high low dose group.Illustrate that below middle dosage (50 μ g/kg body weight), there is dosage one effect dependency relation in bFGF, promptly along with bFGF dosage is increased gradually by 5 μ g → 50 μ g/kg body weight, cerebral infarct volume percent reduces gradually.And escalated dose again, as bFGF250 μ g/kg body weight, its effect and 50 μ g/kg body weight do not have notable difference, promptly are increased to when a certain amount of when bFGF concentration, strengthen bFGF dosage again and effect just no longer occurs and strengthen.The result shows three dosage selecting in the experiment, bFGF50 μ g/kg body weight effect the best.
Embodiment 2
The application of basic fibroblast growth factor in preparation prevention, treatment serious symptom craniocerebral trauma medicine
1, material:
Selected object: the volunteer who gets the heavy craniocerebral trauma that the Shenzhen people's hospital neurosurgery accepts for medical treatment between February in January, 2000 to calendar year 2001 (GCS＜8 minute) is totally 12 examples, male's 10 examples wherein, women's 2 examples, 21～57 years old age, GCS6～8 minute 10 examples, GCS4～5 minute (brain stem injury) 2 examples.Diagnosis: 3 examples of one-sided volume temporal lobe brain contusion companion subdural hematoma operative treatment, brain contusion companion intracerebral hematoma 2 examples of performing the operation, 3 examples of contusion and laceration of brain subarachnoid hemorrhage, 2 examples of motion aphasia appear in one-sided volume temporal lobe brain contusion, a volume temporo subdural hematoma brain contusion left side, right side volume temporo epidural hematoma postoperative companion brain stem injury 1 example, 1 example of extensive brain contusion operation of volume temporo top, right side subdural hematoma and brain stem injury.The limbs muscular strength is measured in treatment previous crops GCS scoring, language function, and a CT scan is to determine position, the scope of brain injury kitchen range.TCD measures bilateral entocranial artery blood circumstance.Medication is front-seat except that after tumor tendency and other complication, implements treatment plan.Therapeutic outcome and conventional therapy group are relatively.
2, Therapeutic Method:
Because bFGF can not pass through blood brain barrier, the present invention selects the common carotid artery administration.For behind the always moving drug administration by injection of neck, understand medicine distribution situation in cerebral tissue, when giving patient ECT scanning, through common carotid artery injection isotope, show that Isotopic Distribution is main in the injection side cerebral hemisphere, prompting is after the common carotid artery injection, and drug main will be distributed in the injection side cerebral hemisphere.In hindering back medication in 2 to 15 days.After the local anaesthesia, the side common carotid artery is hindered in puncture, injects with the quick tremulous pulse of 20% mannitol 50ml, in the hope of opening blood brain barrier better, the common carotid artery perfusion of reuse bFGF80 ten thousand units.The damage of one-sided hemisphere make the homonymy puncture and injection of medicine, the patient of brain stem injury makes the bilateral common carotid arteries puncture and injection of medicine, once a day, totally 10 times.5 days check a CT, ECT, TCD, GCS mark after the medication, and limb function and language function recovery situation remake above-mentioned inspection and contrast with front and back in 10 days, 1 month, 3 months.Same patient's different time injects bFGF after common carotid artery injects 20% mannitol, measure cerebral blood flow velocity before and after the injection with TCD, shows that mannitol can one crosses property cerebral blood flow increasing amount (＞20% about 10 seconds).Illustrate that mannitol can not continue to change the intracranial blood flow, but synergism is arranged with bFGF.
3, result
Patient's 10 examples of GCS6～8 minutes before the treatment, 2 examples of companion's motion aphasia; Contralateral limbs hemiplegia muscular strength I～II is all found in health check-up, and the ETC inspection sees that corresponding brain contusion kitchen range position brain blood is low for difference and cell function, and TCD checks that hindering side cerebral hemisphere homonymy intracranial blood flow velocity speeds.After common carotid artery was with mannitol bFGF, 1～2 day limb function began to recover (muscular strength rises to II～III level by I～II level), and GCS rose to 8～10 fens by 6～8 minutes, and a check CT sees cerebral edema, subarachnoid hemorrhage and occupy-place effect improving after 5 days; TCD checks that the intracranial blood flow slows down before the treatment to some extent; ETC shows that cerebral hemisphere contusion position blood lowly makes moderate progress for the obstacle function of brain cell, medication after 10 days GCS rise to 11～13 fens, muscular strength is more than the IV level, and a CT sees that cerebral edema disappears, and residual softening kitchen range ECT sees that the blood of former edema band is active for recovering function of brain cell; TCD sees that the intracranial blood flow velocity is near normal.Medication after 1 month GCS rise to 14～15 fens, flesh affair IV～V level, a CT see the brain contusion edema residual a small amount of cerebral malacia kitchen range that disappears, ECT sees that a small amount of patchy cell function hangs down inferior segment, TCD examines that into the intracranial blood flow velocity is normal.2 examples have language function obstacle person, begin to recover language function in three months in medication.2 routine brain stem injury persons (GCS4～5 minute), wherein 1 example is diagnosed the diffusing big deep coma of the right volume temporo of right volume temporo hematoma brain contusion epidural hematoma postoperative bilateral pupil, the extremity muscular tension raises, left limb muscular strength III level, the wounded of right side II level, conventional therapy 15 days, the state of an illness does not have improvement, in hindering back beginning in the 16th day through the common carotid artery medication, GCS rises to 6 fens after 5 days, rises to 9 fens after 10 days, muscular tension descends, CT examination bilateral volume temporo cerebral edema begins to disappear, and ECT sees that two volume temporo cerebral blood supply make moderate progress, and TCD sees that bilateral intracranial blood flow velocity slows down to some extent; After the medication 1 month, GCS rises to 14 fens, and muscular tension is normal, left limb muscular strength IV level, and right side III level, language function partly recovers, and TC checks the softening on a small quantity kitchen range of right volume temporo, and ECT sees right frontal lobe blood for slightly poor, and TCD shows that the intracranial blood flow velocity is normal.The recovery from illness of row cranioplasty is left hospital after three months.Another routine brain stem injury is the right volume temporo of a right volume temporo subdural hematoma cerebral hernia postoperative top Big Area Cerebral Infarction, inspection is deep coma, decerebrate rigidity, extremity muscular tension height, right side limbs muscular strength II level, 0 grade in left side, ECT checks a large tracts of land ischemia left side, right volume temporo top volume blood for obstacle, TCD sees that bilateral intracranial blood flow velocity obviously speeds.Through bilateral common carotid arteries perfusion 10 days, the GCS4 branch rises to 7 fens, muscular tension obviously improves, left side human body muscular strength I level, right side II level, check CT sees that the volume temporo pushes up the complete obiteration of left volume low-density kitchen range, and TCD sees that cerebral blood flow speed slows down to some extent, and ECT sees that bilateral frontal lobe cerebral tissue blood is for obstacle, after 1 month, the GCS=9 branch is opened eyes automatically, and muscular tension is slightly high, left side limb holomyarian power II level, right side IV level, a CT check cerebral edema disappears fully and sees the cerebral tissue atrophy, and ECT checks and sees that two frontal lobe patch shape ischemic focus function of brain cell are slightly poor.
Model case one:
Old XX, man, 35 years old; Wounded the head stupor 4 hours by the human fragment of brick, CT scan diagnoses right volume temporo subdural hematoma to merge contusion and laceration of brain, left side volume temporo epidural hematoma, bilateral platycoria before the art, neck rigidity, extremity decorticate rigidity, extremity tension force raises, left limb muscular strength II level, right side III level, left side Babinski sign (ten); Clinical diagnosis: right side subdural hematoma brain contusion, left side epidural hematoma, cerebral hernia, Secondary cases brain stem injury; Row bilateral volume temporo is opened the cranium hematoma clearance lobe decompression of boning, the GCS=5 branch, and CT examination bilateral volume temporo top large tracts of land low-density focus before the art, the bilateral tricorn diminishes.ECT sees that the extensive blood supply disorder function of brain cell of bilateral volume temporo is low, and TCD sees that bilateral entocranial artery blood flow velocity significantly raises.Patient is through routine dehydration, hormone, and the nerve metabolism Drug therapy was still failed improvement in four days, the congested eyeball evagination of conjunctiva of both eyes edema.Through with the present invention's treatment, got the right carotid puncture in the wound back on the 6th day and give 20% mannitol 50ml perfusion back injection bFGF80, ten thousand u, once a day, the bilateral common carotid arteries alternatively administered, GCS rises to 9 fens after 10 days, and ECT sees that left side volume temporo blood supply significantly improves, and right side volume temporo brain blood is for improving.Two volume temporo cerebral edemas of seeing CT bilateral tricorn that disappears is opened again.Left limb muscular strength III level, right side IV level; Patient GCS=14 branch after one month, left limb muscular strength IV level, right side V level, language function partly recovers, and ECT checks that left volume temporo cerebral malacia kitchen range blood is for poor; CT only sees the softening kitchen range brain of the left temporo place of withering.Row cranioplasty after three months, healing is left hospital.
Model case two
Yellow XX, man, 27 years old; Traffic accident causes injury of head, be diagnosed as left volume temporo brain contusion and hard hematoma down, the left volume temporo hematoma clearance of the row lobe decompression of boning, postoperative GCS=8 branch, still there is not improvement through treatment in 5 days, right side limbs muscular strength II level, CT examination volume temporo low-density changes, and ECT checks that left volume temporo large tracts of land blood is low for the obstacle function of brain cell; TCD left side entocranial artery blood flow velocity speeds, and adopts the present invention to treat the 4th day, and GCS rises to 11 fens, and left side muscular strength III level rises to 14 fens after 8 days, motor aphasia occurs, and a check CT sees left volume low-density focus, and ECT shows the left frontal lobe ischemia.TCD sees that left side intracranial offshoot blood flow is roughly normal, and language function is most of after 15 days recovers, and right side limbs muscular strength IV level is left hospital and returned local the rest, makes cranioplasty in three months, and postoperative is worked.
4, result:
The pathophysiological mechanism complexity of secondary brain injury behind the cerebral trauma, but mainly show as cerebral circulation and brain dysbolismus, this group severe cerebral trauma patient, CT show cerebral edema or contusion and laceration of brain, ECT see brain injury position blood for and brain function changes and TCD performance cerebral blood flow velocity such as speeds at form and changing function.Observation index of the present invention is on the clinical manifestation basis, and in conjunction with CT, ECT, TCD check result, index is objective reliable, convincing.The present invention chooses severe cerebral trauma patient and hinders the injection of side cerebral hemisphere common carotid artery, and with the open blood brain barrier of mannitol, being intended to maximum possible increases local drug concentration simultaneously.See in cerebral hemisphere damage (GS≤8 minute) medication 2～3 days that from therapeutic effect of the present invention limb functional recover and the vasodilation of bFGF do relevantly, and the patient of brain stem injury is because of blood vessel maincenter functional disorder.So medication can not be observed the situation of improving blood flow in early days, the sign that could recover to some extent after about 10 days; GCS6-8 based on side cerebral hemisphere damage divides patient, medication after 10 days the function improvement relevant with its refreshing nutrition with new vessels.Language function is a high-level center, and its functional rehabilitation needs injured brain tissue regeneration, and it is relevant promptly to break up, grow with the bFGF induce neural tissue.Brain stem injury person (GCS4～5 minute) is longer because of the time of serious new vessels of its brain injury and neuranagenesis, so that the time that clinical sign and iconography improve appears in medication is longer.Through routine treatment extremely tens of skies of a couple of days, the state of an illness does not have obvious improvement, adds with the present invention to obtain clear and definite curative effect this group patient, and clinical manifestation obviously improves, and the change of iconography is arranged.One routine vitamin K deficiency intracerebral hematoma postoperative hemiplegia is arranged, and after 10 days, the hemiplegia hands can be gone up to lift and grab thing with the bFGF injection for 8 months 1 years old invalid infants of integrative therapy, the present invention's list, and is normal fully after 17 days.After the perfusion of common carotid artery puncture mannitol, inject bFGF, can obviously improve cerebral trauma patient's cerebral circulation and brain metabolism, promote the cerebral trauma rehabilitation of patients, reduce sequela.
Embodiment 3
The application of basic fibroblast growth factor in the preparation anti-cerebral ischemia drugs
(1) experiment material
1. be subjected to reagent product and preparation
(1) basic fibroblast growth factor (bFGF): white lyophilized injectable powder, Bio-engineering Institute of Jinan University provides by Guangzhou, and lot number 981226 is made into medicinal liquid with the volume variable concentrations with bFGF with normal saline during experiment, uses for the rat vein transfusion.
(2) nimodipine is penetrated liquid: colourless transparent liquid, 4mg/20ml (0.2mg/ml) Tianjin people pharmaceutical factory of aminoacid company produces, lot number 990203.
(3) red tetrazolium (TTC): reagent three factories in Shanghai produce, lot number 970410.
(4) rose bengal's disodium (rose red b): Switzerland Fluka company produces.
(5) malonaldehyde (MDA) is measured test kit: Nanjing is built up bio-engineering research and is produced.
2. laboratory animal
(1) adult spontaneous hypertensive rat (SHR):, meet one-level clean animal standard (the moving word of doctor 01-1086 number) available from Fu Wai Hospital, Chinese Academy of Medical Sciences zoopery section.
(2) Wistar rat: Tianjin Inst. of Materia Medica laboratory animal room provides, the animal quality certification: accurate No. 001 of the real moving facility in Tianjin.
3. experimental apparatus
(1) SXP-1B5 type operating microscope, Shanghai medical optical instrument factory.
(2) RM-86 type eight is led physiograph, and Japanese photoelectricity Co., Ltd. can make.
(3) SQ-3 type cerebral thrombosis instrument, unming Medical College makes.
(4) XFH-1 high-speed separation refiner, Shanghai gold reach biochemical instrument factory and produce.
(2) to the influence of focal cerebral ischemia due to blocking-up intraluminal middle cerebral artery occlusion in rats (MCAO)
The Wistar male rat, body weight 291 ± 22g, be divided into 5 groups at random by body weight, 10 every group, lumbar injection 12% chloral hydrate 350mg/kg anesthesia, lateral position is fixed, under operating microscope, cut skin in external auditory meatus and eye corner of the eyes line mid point, expose frontal bone, hinge is disconnected, open about 2mm diameter microcephalia window in the skull bottom, expose middle cerebral artery, provoke with pin, break it, fill hemostasis with sthptic sponge immediately, treat blood clotting after, the local 2-3 that drips drips penicillin (160,000 units/ml), in case wound infection, layer-by-layer suture.The different medicinal liquids of operation back 30min microinfusion pump vein constant speed gasing injection; Matched group infusion normal saline, the administration group is infusion bFGF10,20,40ug/kg respectively, and every Mus administration volume is 3ml, and the infusion time is 3h; Positive controls nimodipine 10ug/kg behind MCAO 0.5, each intravenous injection of 2h once, postoperative 24h carries out the animal behavior scoring, with this as the disordered brain function index.Blind method is adopted in scoring, and promptly scoring person does not know to give the situation of medicine.Standards of grading such as table 2:
Table 2 disordered brain function index
Symptom disordered brain function index score
Carry the about chi of Mus tail built on stilts, the wrist flexing appears in operation offside forelimb,
The elbow flexing is takeed on inward turning or wrist is arranged, and the elbow flexing has inward turning, 1-4 again
Animal is placed on the plane earth, push away both shoulders respectively, check resistance to offside.1-3
Animal is placed on the wire netting, observe the tension force of two forelimbs.1-3
Animal is placed on the plane earth, observe to have or not and turn-take.1-2
Full marks 12
Above standard full marks are 12 minutes, and mark is high more, show that the animal behavior obstacle is serious more.
Scoring finishes broken end and gets brain, and the visible control animals focus of perusal side brain surface is pale, unglazed, and the cerebral tissue edema is obvious; The most of animal edema of administration group are light than matched group, and the focus brain is pale unglazed not obvious, then brain is put into normal saline and handle, only keep the bilateral cerebral hemisphere and weigh, be cut into 5 along coronalplane, brain section is put into the 1%TTC dye liquor, 37 ℃ of lucifuge temperature are incubated 30min, and it is rose-colored that normal structure is, and infarct is white in color, blocking part cut weigh, calculate blocking tissue's weight with " weight area method " and account for the heavy percentage ratio of bilateral cerebral hemisphere as infarction size.The results are shown in Table 3,4.
Table 3 bFGF is to the shadow of local rats with cerebral ischemia behavior (X ± SD)
Group dosage (ug/kg) number of animals (only) behavior disorder (24h)
Sham operated rats-10 0 ± 0
Model control group 0.5ml/100g 10 7.9 ± 1.33
bFGF 10 10 4.65±2.16
***
bFGF 20 10 4.55±1.86
***
bFGF 40 10 3.45±2.01
***
Nimodipine 10 * 2 10 2.85 ± 1.45
* *
Compare with model control group:
* *P＜0.001.
As can be seen from Table 3, the sham operated rats animal behavior does not have obvious change, model control group animal and sham operated rats animal are relatively, tangible behavior disorder appears, rat vein infusion bFGF 10,20,40ug/kg, its behavior variation and matched group relatively have clear improvement, and behavior scoring has reduced by 41.1% respectively behind the 24h, 42.4%, 56.3% (p is all＜0.001 〉.The effect of positive drug nimodipine is obvious.
Table 4bFGF is to the influence of local rats with cerebral ischemia cerebral infarct size (X ± SD)
Group dosage (ug/kg) number of animals (only) infarction size (%)
Sham operated rats-10 0.0
Model control group 0.5ml/100g 10 6.92 ± 1.83
bFGF 10 10 4.48±3.14
*
bFGF 20 10 3.98±2.33
**
bFGF 40 10 1.76±0.77
***
Nimodipine 10 * 2 10 1.92 ± 1.18
* *
Compare with model group:
*P＜0.05,
*P＜0.01,
* *P＜0.001.
Table 4 is the result show, relatively infarction size is obvious for model group animal and sham operated rats animal.Behind venoclysis BFGF10,20, the 40ug/kg, can obviously dwindle the rats with cerebral ischemia cerebral infarct size, more on average dwindled 35.3% with matched group (P＜0.05 〉, 42.5% (P＜0.01 〉, 74.6% (P＜0.001 〉, and increase infarction size with dosage and dwindle, the effect of positive drug nimodipine is obvious.
(3) to the influence of ligation bilateral common carotid arteries salt load Hypertensive Rats mortality rate
Select body weight 312 ± 64g original hypertensive rat (SHR) for use, the male and female dual-purpose, indirect method is measured the rat systolic pressure, by body weight, blood pressure is divided into 5 groups at random, 10 every group: matched group, bFGF10,20, the 40ug/kg group, nimodipine 10ug/kg group, each treated animal 1% sodium chloride of feeding begins experiment after 5 days, rat is with 12% chloral hydrate 350mg/kg intraperitoneal injection of anesthesia during experiment, animal faces upward the position and is fixed on the operating-table, separate bilateral common carotid arteries, with No. 4 silk thread ligation, skin suture, 30min difference femoral vein infusion different pharmaceutical after the ligation, the administration volume is every Mus 2ml, and the time is 1h, nimodipine be postoperative 30min intravenous injection once, inject once again behind the 15h, administration finishes the back animal and steams again raising, observes death toll behind the 24h, and experimental result sees Table 5.
Table 5 bFGF is to the influence of the total tremulous pulse salt load of ligation bilateral Hypertensive Rats mortality rate
Group dosage (ug/kg) number of animals (only) death toll X
2Value P value
Matched group-10 9--
bFGF 10 10 6 1.067 ＞0.05
bFGF 20 10 4 3.516 ＞0.05
bFGF 40 10 3 5.208 ＜0.05
Nimodipine 10 * 2 10 3 5.208＜0.05
Above result shows, venoclysis bFGF10,20,40ug/kg all increase ligation bilateral common carotid arteries salt load Hypertensive Rats survival rate, effect of high dosage obviously (P＜0.05 〉.The effect of positive drug nimodipine is obvious.
(4) to the diffusivity imperfection cerebral ischemia reperfusion MDA of rat cerebral tissue content influence
Body weight 313 ± 31gWistar male rat, be divided into 6 groups at random by body weight, every group 10, lumbar injection 20% urethane 1g/kg anesthesia, facing upward the position is fixed on the operating-table, separate the usefulness of bilateral common carotid arteries threading in order to blocking-up, by the external jugular vein intubate to right atrium, reach filling again in order to get blood, the right femoral artery intubate, blood pressure signal is measured blood pressure (MABP) through MPU-0.5 type pressure transducer input RPF-5 carrier amplifier, blood pressure is led the physiograph record by eight, write down normal arterial pressure during the experiment beginning earlier,, make blood pressure reduce to 80mmHG then from the external jugular vein blood-letting, clamp bilateral common carotid arteries with bulldog clamp this moment, continue to make blood pressure to reduce to 50mmHg, and keep 30min, this is an Ischemia Time.Behind the ischemia 15min, beginning femoral vein infusion bFGF20,40,80ug/kg, model control group infusion normal saline, positive drug infusion nimodipine 30ug/kg, the administration volume is every Mus 2ml, time is unclamped the bilateral common carotid arteries clip after being 1h ischemia 30min, and the blood that will take out is irritated in the ex vivo by external jugular vein, 1.5h after with sacrifice of animal, get Hippocampus and cortex, weigh, it is freezing standby to put into refrigerator.
MDA assay: get the freezing brain of refrigerator, make brain homogenate with normal saline and 1: 9 ratio of cerebral tissue on XFH-1 high-speed separation refiner, then 4000 rev/mins of centrifugal 10min get supernatant, adopt the TBA method to measure MDA content, adopt biuret method to carry out quantification of protein and measure.The results are shown in Table 6.
Table 6 bFGF is to the influence of global brain ischemia rat MDA content (X ± SD)
Group dosage (ug/kg) number of animals (only) MDA content (nmol/mgpo)
Normal control group-10 35.95 ± 6.36
*
Model control group 0.5ml/100g 10 45.66 ± 5.25
bFGF 20 10 47.09±2.44
bFGF 40 10 41.54±5.31
bFGF 80 10 38.59±8.34
*
Nimodipine 30 10 36.32 ± 4.55
* *
Compare with model control group:
*P＜0.05,
*P＜0.01,
* *P＜0.001.
Above result shows that model contrast cerebral tissue MDA content is apparently higher than normal control group (P＜0.01).Give global brain ischemia re-perfusion model rat bFGF80ug/kg, can obviously reduce the MDA content (comparing P＜0.05 with model control group) in its cerebral tissue, show that bFGF has obvious protective effect to cerebral ischemia, the positive drug nimodipine also obviously reduces MDA content.
(5) to the influence of photochemical induction rat suppository cerebral ischemia
Select body weight 219 ± 41gWistar male rat for use, body weight 219 ± 41g is divided into 6 groups by body weight, every group 10, lumbar injection 12% chloral hydrate 350mg/kg, the ventricumbent position is fixed, cut left top scalp, expose skull,, animal is put " cerebral thrombosis experimental provision " in the subcutaneous barrier sheet (fenestra that contains 0.5 * 0.6cm) of putting, inject the normal saline solution 0.133ml/100g that contains 0.75% brave red B from sublingual vein, (light intensity is 0.5w/cm to make light source behind the 5min
2) by parietal bone surface, fenestra direct irradiation left side, irradiation time 20min, illumination finishes back 30min, femoral vein infusion bFGF20,40,80ug/kg, matched group infusion normal saline, positive drug group nimodipine 40ug/kg, each medicine volume is every Mus 2ml, the infusion time is 1h, puts to death animal behind the 24h, and it is clean with normal saline flushing to get forebrain, and blot with filter paper, be divided into left and right brain hemisphere, get left hemisphere and put clean vial, 100 ℃ of 24h dry to constant weight.Brain water content calculates by " (wet amount-dry weight)/weight in wet base=water % ", and experimental result sees Table 7.
Table 7bFGF is to the influence of optical induction rat suppository cerebral ischemia brain water content (X ± SD)
Group dosage (ug/kg) number of animals (only) brain water content (%)
Sham operated rats-10 78.79 ± 0.29***
Model control group 0.5ml/100g 10 79.62 ± 0.52
bFGF 20 10 79.34±0.55
bFGF 40 10 79.04±0.24**
bFGF 80 10 78.99±0.36**
Nimodipine 40 10 79.00 ± 0.12**
Compare * * P＜0.01, * * * P＜0.001. with model control group
Above experimental result shows, photochemically-induced rat suppository cerebral ischemia cerebral edema water content is apparently higher than sham operated rats (P＜0.001), and venoclysis bFGF40,80ug/kg can obviously alleviate photochemical induction rat suppository cerebral ischemia cerebral edema (P＜0.01).The effect of positive drug nimodipine is obvious.
(6) conclusion
1, venoclysis bFGF10,20,40ug/kg can obviously improve blocking-up middle cerebral artery focal cerebral ischemia rat nervous symptoms, and can reduce its infarction size.
2, venoclysis bFGF40ug/kg can obviously reduce ligation bilateral common carotid arteries salt load Hypertensive Rats mortality rate.
3, venoclysis bFGF80ug/kg can obviously reduce global brain ischemia and irritates the MDA of rat cerebral tissue content again.
4, venoclysis bFGF40,80ug/kg can obviously alleviate photochemical induction rat suppository cerebral ischemia cerebral edema.
Claims (4)
1, the application of basic fibroblast growth factor in preparation prevention, treatment brain injury medicine.
2, application as claimed in claim 1 is characterized in that brain injury is the damage that cerebral trauma, apoplexy, nerve regression disease or brain operation are brought.
3, application as claimed in claim 2 is characterized in that apoplexy is cerebral ischemia or cerebral hemorrhage.
4, application as claimed in claim 2 is characterized in that nerve regression disease is senile dementia or Parkinson's disease.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB011147563A CN1150944C (en) | 2001-05-31 | 2001-05-31 | Application of alkaline fibroblast growth factor in preparing medicine to prevent and cure cerebral injury |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB011147563A CN1150944C (en) | 2001-05-31 | 2001-05-31 | Application of alkaline fibroblast growth factor in preparing medicine to prevent and cure cerebral injury |
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| CN1325730A CN1325730A (en) | 2001-12-12 |
| CN1150944C true CN1150944C (en) | 2004-05-26 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101575602B (en) * | 2008-05-06 | 2011-06-22 | 天津市第一中心医院 | siRNA sequence for inhibiting human BFGF gene expression amplification and expression carrier and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108421033A (en) * | 2018-03-12 | 2018-08-21 | 杭州荣泽生物科技有限公司 | The reagent and its application method of promotion nerve stem cell proliferation based on ginsenoside Rg1 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101575602B (en) * | 2008-05-06 | 2011-06-22 | 天津市第一中心医院 | siRNA sequence for inhibiting human BFGF gene expression amplification and expression carrier and application thereof |
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