CN110179808A - Application of the Gastrodin in preparation treatment cerebral hemorrhage drug - Google Patents
Application of the Gastrodin in preparation treatment cerebral hemorrhage drug Download PDFInfo
- Publication number
- CN110179808A CN110179808A CN201910418762.6A CN201910418762A CN110179808A CN 110179808 A CN110179808 A CN 110179808A CN 201910418762 A CN201910418762 A CN 201910418762A CN 110179808 A CN110179808 A CN 110179808A
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- gastrodin
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- cerebral
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Classifications
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
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- A61K38/49—Urokinase; Tissue plasminogen activator
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Abstract
The present invention relates to pharmaceutical technology fields, and in particular to purposes of the Gastrodin in preparation cerebral hemorrhage drug especially reduces the purposes after cerebral ischemia rt-PA thromboembolism treatment in caused hemorrhagic conversion drug in preparation.The present invention provides Gastrodins to reduce the purposes after cerebral ischemia in hemorrhagic conversion drug caused by rt-PA thromboembolism treatment in preparation, prompt is clinical as early as possible using Injectio of gastrodine for using rt-PA thromboembolism treatment ischemic cerebral apoplexy and existing hemorrhagic conversion will play certain inhibiting effect, opposite it can extend rt-PA thrombolytic treatment time window, strive for more treatment times for clinical patients with cerebral apoplexy, further decrease the injury of Central nervous system, so that favourable prognosis is reduced sequelae, improves patients ' life quality.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to purposes of the Gastrodin in preparation cerebral hemorrhage drug, especially
The purposes after cerebral ischemia rt-PA thromboembolism treatment in caused hemorrhagic conversion drug is reduced in preparation.
Background technique
It is well known that intravenous thrombolysis be treatment the only effective method of acute ischemic cerebral apoplexy, safety and effectively
Property gained public acceptance, and by multinational guide recommend [Hacke W, Donnan G, Fieschi C, et al.Association of
Outcome with early stroke treatment:pooled analysis of ATLANTIS.ECASS.and
NINDS rt-PA stroketrials [J] .Lancet, 2004,363 (9411): 768-774.].Currently, domestic and international guide one
It causes to recommend rt-PA (rt-PA) intravenous thrombolysis to be to treat the best side of acute ischemic cerebral apoplexy
Case, and treatment is even more thrombolysis safety and the guarantee of validity [Jauch EC, Saver JL, Adams HP in the 4.5h that falls ill
Jr,et al.Guidelines for the early managerment of patients with acute ischemic
stroke:A guideline for healthcare professionals from American Heart
Association/American Stroke Association[J].Stroke,2013,44:870-947.].However vein
Thrombolysis increases the risk of Hemorrhagic Transformation (hemor-rhagic transformation, HT), and HT is cerebral arterial thrombosis
Common severe complication dramatically increases the disability rate and the death rate of stroke especially after thromboembolism treatment, is that thromboembolism treatment is most main
Will, most dangerous complication, conditions of patients can be caused sharply to deteriorate and dead, affect the efficacy and saferry of thrombolysis.Tissue
Plasminogen activator is that is be approved by the FDA in the United States be uniquely used for the thrombolytic drug of cerebral apoplexy, since therapeutic time window is narrow and easy
Cause hemorrhagic conversion, limits its clinical application.Although the incidence of HT is lower, the risk of this complication is likely to become
People make heavy burden when thromboembolism treatment determines, hinder the popularization of thromboembolism treatment.
The pathomechanism of HT and therapy target are unclear at present, it is clinical without effectively prevent drug (Chen Yanxia, Kong Linglei,
Effect [J] the Acta Pharmaceutica Sinica of, king harbour etc. Nuo Sailin in the hemorrhagic conversion caused by t-PA thrombolysis, 2019,54 (3): 448-
453)。
Therefore how to overcome cerebral hemorrhage caused by rt-PA；How the curative effect of acute ischemic cerebral apoplexy thromboembolism treatment is improved
And safety, to reduce headstroke disable and lethality has great importance.
Gastrodin has calm, soporific function, has stronger protective effect to neurotrosis, can significantly reduce nerve cell
The death rate, have relaxation effect to neurasthenia, insomnia, headache syndromes；In addition, also having free radical resisting, anti-inflammatory, anti-oxidant etc.
Pharmacological action.
So far, there is not yet related Gastrodin after reducing acute ischemic cerebral apoplexy rt-PA thromboembolism treatment for causing
Hemorrhagic conversion report.
In view of this present invention is specifically proposed.
Summary of the invention
Go out the technical problem to be solved in the present invention is that overcoming the deficiencies of the prior art and provide a kind of Gastrodin preparing brain
Purposes in blood drug, especially the Gastrodin caused hemorrhagic conversion drug after preparation reduces cerebral ischemia rt-PA thromboembolism treatment
In purposes.
In order to solve the above technical problems, the present invention adopts the following technical scheme:
The present invention provides application of the Gastrodin in preparation treatment cerebral hemorrhage drug, wherein the cerebral hemorrhage lacks for brain
Application after blood rt-PA thromboembolism treatment in caused hemorrhagic conversion drug.
Intravenous thrombolysis is the treatment the only effective method of acute ischemic cerebral apoplexy, and rt-PA intravenous thrombolysis is that treatment is anxious
The preferred plan of property cerebral arterial thrombosis.However intravenous thrombolysis increases Hemorrhagic Transformation (hemor-rhagic
Transformation, HT) risk, HT is the complication that thromboembolism treatment is main, most dangerous, can cause conditions of patients sharply
Deteriorate and dead, affects the efficacy and saferry of thrombolysis.Although the incidence of HT is lower, the risk of this complication can
Heavy burden when people make thromboembolism treatment decision can be become, hinder the popularization of thromboembolism treatment.Therefore how to overcome rt-PA
Caused cerebral hemorrhage；The efficacy and saferry for how improving acute ischemic cerebral apoplexy thromboembolism treatment, the cause to headstroke is reduced
Residual and lethality has great importance.
The present inventor is by a large amount of research discovery Gastrodin and rt-PA Drug combination, it is possible to provide effectively fills again
Note, prevents from intracranialing hemorrhage, reduces disability rate, the lethality of acute ischemic cerebral apoplexy, opposite to extend rt-PA thrombolytic treatment time
Window makes more patients obtain Normalized Treatment.The present invention is further study showed that Gastrodin joint rt-PA treats acute cerebral infarction
The mechanism of hemorrhagic conversion caused by dead may refer to the serum inflammatories such as MMP-9 activation, downward IL-1 β, IL-6, IL-12, TNF α are inhibited
Mark, which is overexpressed, has certain correlation, and Gastrodin raises TJs correlative protein expression level simultaneously, maintains TJs structure and function steady
It is qualitative to play cerebral protection.
Therefore, the present invention provides purposes of the Gastrodin in preparation cerebral hemorrhage drug, and especially Gastrodin is reduced in preparation
Purposes after cerebral ischemia rt-PA thromboembolism treatment in caused hemorrhagic conversion drug.
Further, caused hemorrhagic conversion is that cerebral ischemia rt-PA thrombolysis is controlled after the cerebral ischemia rt-PA thromboembolism treatment
Hemorrhagic conversion caused by MMP-9 increased activity after treatment.
Further, caused hemorrhagic conversion is that cerebral ischemia rt-PA thrombolysis is controlled after the cerebral ischemia rt-PA thromboembolism treatment
Hemorrhagic conversion caused by IL-1 β, IL-6 and/or IL-12 are overexpressed after treatment.
Further, caused hemorrhagic conversion is that cerebral ischemia rt-PA thrombolysis is controlled after the cerebral ischemia rt-PA thromboembolism treatment
Hemorrhagic conversion caused by TNF α expression enhances after treatment.
Further, caused hemorrhagic conversion is that cerebral ischemia rt-PA thrombolysis is controlled after the cerebral ischemia rt-PA thromboembolism treatment
Hemorrhagic conversion caused by TJs correlative protein expression level declines after treatment.
The present invention also provides Gastrodin prepare in MMP-9 activity inhibitor drug application, preparation IL-1 β, IL-6,
In IL-12 expression inhibiting agent drug application and preparing the application in TNF α expression inhibiting agent drug.
The present invention is verified by experiments, and Gastrodin can be overexpressed by lowering MMP-9, while reduce IL-1 β, IL-6, IL-
12, the expression of the inflammatory factors such as TNF α, rt-PA thrombolysis is to cerebrovascular destruction after effectively blocking cerebral ischemia, to reduce
The cerebral hemorrhage that rt-PA thrombolysis induces.
The invention shows pass through close connection (TJs) GAP-associated protein GAP of up-regulation: tight junction protein -5 (Claudin-5) and
Closed protein (Occludin) mRNA transcription and protein expression level, Gastrodin have certain protective effect to TJs and BBB.
Present invention prompt, Gastrodin are reducing brain tissue bleeding wind caused by clinical use rt-PA thromboembolism treatment post-stroke
Survival etc. is improved with good development and application values in danger, it may be possible to prevent and treat the potential of HT caused by rt-PA thrombolysis
Drug.Therefore, Gastrodin is used in combination and rt-PA is a kind of novel countermeasure and means for increasing thrombolysis safety.
Further, drug of the present invention further includes pharmaceutically acceptable auxiliary material.
It is preferred that the dosage form of the drug is injection, tablet, capsule, pulvis, pill or oral solution.
The present invention also provides a kind of combination product containing the first medicament and second medicament, wherein first medicament is rt-
PA, the second medicament are Gastrodin.
In the present invention, the first medicament rt-PA is common rt-PA in the market；Second medicament Gastrodin can be this field
Common includes the medicament of Gastrodin, is also possible to Gastrodin and pharmaceutically acceptable auxiliary material according to pharmaceutically common system
Pharmaceutically acceptable dosage form, optimizing injection, tablet, capsule, pulvis, pill or oral solution is made in Preparation Method.
The present invention furthermore provides the composition product in preparation treatment acute ischemic cerebral apoplexy drug
Using.
Rt-PA is as the thrombolytic drug for being uniquely used for cerebral apoplexy by FDA approval, when clinical application is limited to relatively narrow treatment
Between window and high hemorrhagic conversion (HT) incidence.Therefore, clinical application of the new HT protective agents for expansion rt-PA is found, is changed
The prognosis of kind thrombolytic therapy has great importance.The research of the invention finds that there are reperfusion injuries for thrombolysis process, it is adjoint always
Oxidative stress generates a large amount of inflammatory factor reactions, and it is anti-to generate a series of cascades such as a large amount of free radicals and promotion MMP-9 level raising
It answers, nervus vasculairs unit is caused to damage, destroy blood-brain barrier (BBB) or even hemorrhagic conversion.Gastrodin can reduce scarce to a certain degree
Blood side brain tissue MMP-9 is horizontal, and reduces the serum inflammatory parameters such as IL-1 β, IL-6, IL-12, TNF α, illustrates that Gastrodin may
Brain tissue hemorrhagic conversion is induced to brain tissue impairment caused by ischemia-reperfusion and rt-PA by anti-inflammatory, inhibition oxidative stress
Protective effect is played, the BBB for inhibiting rt-PA to mediate is destroyed and the generation of HT.Inside and outside combination research institute obtain the result shows that, rt-
PA use in conjunction Gastrodin reduces ischemic side brain tissue hemoglobin level in various degree, and mortality of animals is decreased obviously, shows
Gastrodin can protect BBB integrality and reduce the generation of HT；The opposite thromboembolism treatment window for extending rt-PA, for clinical brain soldier
Middle patient strives for more treatment times.These results indicate that Gastrodin and rt-PA combination can fight acute ischemic cerebral apoplexy
The adverse reaction of thromboembolism treatment inhibits the generation of HT, provides new thinking for the treatment of acute ischemic cerebral apoplexy.
Combination product of the invention is when being used to treat acute ischemic cerebral apoplexy, the first medicament rt-PA and second medicament
Gastrodin can be used simultaneously, and first can also reuse Gastrodin using rt-PA, first can also reuse rt-PA using Gastrodin.Make
It is first to reuse rt-PA using Gastrodin for a kind of preferred embodiment.Studies have shown that treating acute ischemic brain using rt-PA
Gastrodin is first used before stroke, can substantially reduce the generation of HT.
After adopting the above technical scheme, compared with the prior art, the invention has the following beneficial effects:
The mechanism of hemorrhagic conversion caused by Gastrodin joint rt-PA treatment acute cerebral infarction of the present invention may be with inhibition MMP-9
Activation is lowered IL-1 β, IL-6, IL-12, the serum inflammatory parameters such as TNF α and is overexpressed and has certain correlation, Gastrodin simultaneously on
It adjusts TJs correlative protein expression horizontal, TJs structure and function stability is maintained to play cerebral protection.
A specific embodiment of the invention is described in further detail with reference to the accompanying drawing.
Detailed description of the invention
Fig. 1 is each group cerebral infarction percentage；
Fig. 2 is each group brain water content percentage；
Fig. 3 is neurological deficit score；
Fig. 4 is the scoring of brain tissue hemoglobin level；
Fig. 5 is groups of animals brain tissue extent of hemorrhage；
Fig. 6 is brain tissue bleeding ratio and animal dead rate dependence；
Fig. 7 is blood coagulation four indices；
Fig. 8 is each group brain tissue MMP-9 content；
Fig. 9 is blood inflammatory cytokines levels in non-diabetic；
Figure 10 is changes of weight percentage；
Figure 11 be OGD induction HBMEC damage different time after cell survival rate (N=6)；
Figure 12 is the Injectio of gastrodine of various concentration to cell survival rate after OGD induction HBMEC damage different time
Influence (N=6)；
Figure 13 be group of cells related gene mRNA transcription situation (N=3)；
Figure 14 is Injectio of gastrodine to OGD induced damage HBMEC Claudin-5, Occludin and ZO-1 protein expression
Horizontal influence；
Figure 15 be group of cells Claudin-5, Occludin and ZO-1 protein expression situation (N=3).
It should be noted that these attached drawings and verbal description are not intended to the design model limiting the invention in any way
It encloses, but illustrates idea of the invention by referring to specific embodiments for those skilled in the art.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, the technical solution in embodiment is clearly and completely described, the following examples are intended to illustrate the invention, but
It is not intended to limit the scope of the invention.
Embodiment 1: Gastrodin causes the research of rats after cerebral ischemic reperfusion hemorrhagic conversion protective effect to rt-PA
1. reagent and animal
1.1 test samples: Injectio of gastrodine, Kun Yao Group Plc, character: colourless transparent liquid；Specification:
2mL:0.2g/ branch；Lot number 17JX06-222；Preservation condition: room temperature is protected from light；Validity period: 2020.09；
1.2 reference substances: title or code name: Alteplase (rt-PA)；German Boehringer Ingelheim company (Germany)；Character:
The loose block of off-white color；Specification: 20mg/ branch (contains dilution)；Lot number: 703948；Preservation condition: it is 2~8 DEG C, closed, it keeps away
Light；Validity period: 2020.02；
1.3 other reagents: quantitative hemoglobin kit (BioAssay Systerms, the U.S., Cat:DIHB-250)；
MMP9 ELISA detection kit (R&D, the U.S., Cat:RMP900)；Rat Cytokine/Chemokine (TNF-α, IL-6,
IL-12 (p70), IL-1beta) liquid-phase chip, manufacturer: Millipore, the U.S., Cat:RECYTNAG-65K.
1.4 experimental animal
SD rat: SPF grades, male, 180, weight: 235.58~329.46g, weight individual values are in ± 20% model of mean
In enclosing；Beijing Vital River Experimental Animals Technology Co., Ltd., production licence number: SCXK (capital) 2016-0011.Experimental animal
20~26 DEG C of room temperature (temperature difference per day≤4 DEG C), humidity 40%~70%, illumination 12h:12h light and shade alternating, illumination 150~
300lx, noise≤60Db；Stocking density :≤5/cage, feeding environment condition standard: People's Republic of China's national standard
GB14925-2010；Feeding environment control system: Honeywell company EBI400 automatically controls all-fresh air central air conditioner system；
Experimental animal uses licensing: SYXK (river) 2013-009.Daily feeding mouse mixed feed, free water are optionally replaced
Cage tool and padding；Feed resource examines the quality certification: Shanghai in Shanghai Slac Experimental Animal Co., Ltd., Feedstuff Enterprises
Raise card (2014) 04001.
2. experiment content
2.1 animal packet
Group design: sham-operation group, solvent control group, rt-PA group, Gastrodin group, Gastrodin (- 60min)+rt-PA group,
Rt-PA+ Gastrodin (0h) group, rt-PA+ Gastrodin (60min)；
Group technology: it after rat MCAO modeling, is randomly assigned according to group district's groups.
2.2 dose design
It is shown in Table 1-1.
Table 1-1. dose design table
2.3 administration
Administration route: tail vein injection injects time about 2min；Administration frequency: single-dose；Administered volume: 5mL/kg
2.4 test method
Intraluminal middle cerebral artery occlusion in rats is caused to occlude (MCAO) Focal Cerebral Ischemia Reperfusion by line brush, after ischemic two hours
Pull out Outlet bolt realize physics fill (solvent control group) again, separately take one group before multiple filling 10min give rt-PA analog drug physical property thrombolysis
Lead to (rt-PA group) again.
Surgical procedure is as follows:
Rat is maintained to anaesthetize with isoflurane, is lain on the back, after neck median incision, blunt separation muscle layer, exposure right side
Arteria carotis communis successively isolates outer neck, internal carotid and other relevant blood vessel branches.By external carotid artery from the twice silk thread of ligation
Centre is cut, and holding stump makes external carotid artery and internal carotid in a straight line, makees notch in external carotid artery, by what is be marked
Line bolt (model: 2838-A4) is slowly promoted to internal carotid, until slightly (fishing line front end is from intersection inside and outside neck for sense resistance
About 18mm).After ischemic 2 hours, suture is unlocked, line bolt is extracted completely, tightens artery stump, realizes Reperfu- sion, rt-PA
Then 10min gives rt-PA (5mg/kg) to group before pulling out line bolt.Cut off extra silk thread, skin suture.Rat is maintained in surgical procedure
35-37 DEG C of body temperature.For rats in sham-operated group in addition to being not inserted into line bolt, other operate same solvent control group.
2.5 observation index
2.5.1 general state is observed
Observing time and frequency: 24 hours each 1 time after the administration same day and Reperfu- sion；
Observe result: administration phase animal subject integrality, including but not limited to administration local reaction.
2.5.2 neurological deficit score
It scores the time: nervous function damage neurological deficit score being carried out to modeling animal before administration；
Methods of marking: (0 point: impassivity injury symptoms of 5 point-scores；1 point: being unable to full extension opposite side fore paw；2 points: outward
It turn-takes；3 points: being tilted to opposite side；4 points: it spontaneous cannot walk, the loss of consciousness), scoring divides rat to be determined as modeling qualification in 1-3,
It is administered.
2.5.3 24 hours mortality of animals of MCAO Reperfu- sion
24 hours groups of animals death conditions after MCAO Reperfu- sion are observed and recorded, the death rate is calculated.
The death rate=each group dead animal quantity/each group experimental animal quantity × 100%.
2.5.4 24 hours brain tissue Bleeding rates of MCAO Reperfu- sion
24 hours after MCAO Reperfu- sion, each group survival rats are dissected, take brain tissue after heart perfusion using PBS.Observation
And groups of animals brain tissue ischemia side bleeding is recorded, calculate brain tissue Bleeding rate.
Brain tissue Bleeding rate=brain tissue bleeding size of animal/surviving animals quantity × 100%.
2.5.5 brain tissue content of hemoglobin
24 hours after MCAO Reperfu- sion, cerebral tissue on the right side of each group partial rat is taken respectively, total protein is extracted, according to blood red
Protein quantification kit specification detects content of hemoglobin.
2.5.6 cerebral infarction volume
24 hours after MCAO Reperfu- sion, each group survival rats are dissected, every group selection part surviving animals take brain tissue, use
1% red tetrazolium (TTC) is dyed, and Infarction volume percentage is calculated.
Cerebral infarction volume percentage=right side Infarction volume/big brain volume × 100% in right side.
2.5.7 coagulation indexes
24 hours after MCAO Reperfu- sion, each group survival rats are dissected, take blood about 2.0mL, sodium citrate is anticoagulant.4 DEG C,
3000rpm is centrifuged 10min, separated plasma.Detect coagulation indexes: PT, APTT, TT, FIB.
2.5.8 brain tissue MMP9 content
24 hours after MCAO Reperfu- sion, cerebral tissue on the right side of each group partial rat is taken respectively, extracts total protein.According to explanation
Book detects MMP9 content.
2.5.9 serum inflammatory factors of senile
24 hours after MCAO Reperfu- sion, each group survival rats are dissected, take blood about 3.0mL, sodium citrate is anticoagulant.4 DEG C,
3000rpm is centrifuged 10min, separates serum, and Luminex detects TNF-α, IL-1 β, IL-6, IL-12 content.
2.5.10 weight
Operation consent and a weight of animals is respectively weighed after Reperfu- sion 24 hours.
3. statistical analysis
Test data is indicated with Mean ± SEM, examines (M-W method) to compare group difference using Mann-Whitney U
Compared with.Results of statistical analysis P < 0.05 thinks there is significant difference, and P < 0.01 thinks there is extremely significant sex differernce.
4. test result
4.1 cerebral infarction volume
After MCAO Reperfu- sion 24 hours, Gastrodin is applied alone compared with solvent control group, and cerebral infarct volume mean value is substantially reduced.
Gastrodin group is substantially reduced earlier than rt-PA 60min administration cerebral infarct volume mean value compared with rt-PA group, sees Fig. 1.
4.2 brain water content
After MCAO Reperfu- sion 24 hours, solvent control group and the extremely significant property of rt-PA group brain water content are higher than sham-operation group
(**P < 0.01).For Gastrodin group compared with solvent control group, brain water content is on a declining curve.Compared with rt-PA group, Gastrodin exists
60min is administered before rt-PA, and brain water content has downward trend to see Fig. 2.
4.3 neurological deficit score
Before MCAO administration, neurological deficit score is carried out, each modeling group rat behavior scoring mean value is suitable, without significance difference between group
It is different.24 hours after multiple filling, Gastrodin (120mg/kg) is applied alone and different time is administered before and after giving rt-PA, neurological deficit score
There is different degrees of decreasing trend, wherein 60min gives Gastrodin animal movement function ratio rt-PA group and is obviously improved before rt-PA
(aP < 0.05), see Fig. 3.
4.4 brain tissue content of hemoglobin
After MCAO Reperfu- sion 24 hours, rt-PA (5mg/kg) organizes the apparent increase of rat cerebral tissue's hemoglobin level, significantly
Higher than solvent control group (*P < 0.05).Compared with rt-PA (5mg/kg) is applied alone, day is given giving the forward and backward 60min of rt-PA
Numb element (120mg/kg) reduces ischemic side brain tissue hemoglobin level in various degree and sees Fig. 4.
The 4.5 brain tissue Bleeding rates of MCAO Reperfu- sion 24 hours
After 24 hours, rt-PA (5mg/kg) administration group rat cerebral tissue bleeding ratio is apparently higher than rat MCAO Reperfu- sion
Solvent control group, and severity obviously increases.Compared with rt-PA is applied alone, Gastrodin is given giving rt-PA different time points
(120mg/kg), rat cerebral tissue's bleeding ratio is on a declining curve, and bleeding severity is mitigated, and sees Fig. 5.Correlation analysis
It has been shown that, 24 hours mortality of animals of MCAO Reperfu- sion and brain tissue bleeding ratio are in strong correlation (r=0.92, P=0.0033), such as
Fig. 6, prompting bleeding after rt-PA induced rat MCAO ischemia-reperfusion is to lead to the principal element of animal dead.
4.6 mortality of animals of MCAO Reperfu- sion 24 hours
For rat MCAO Reperfu- sion after 24 hours, the solvent control group death rate is 30.4% (7/23), rt-PA (5mg/kg) group
The death rate is 52% (13/25), and the rt-PA group death rate is apparently higher than solvent control group.Compared with solvent control group, individually give
Gastrodin (120mg/kg) death rate (12%) improves significant；Compared with rt-PA is applied alone, 60min gives day before giving rt-PA
Numb element (120mg/kg), mortality of animals is decreased obviously；Gastrodin, animal dead are given simultaneously and after 60min giving rt-PA
Rate is on close level with rt-PA group, is shown in Table 1-2.
24 hours death rates after table 1-2 MCAO Reperfu- sion
4.7 coagulation indexes
After MCAO Reperfu- sion 24 hours, solvent control group and rt-PA group APTT dramatically increase than sham-operation group (*P <
0.05), rt-PA group APTT is higher than solvent control group, such as Fig. 7 A.Gastrodin is in the administration of rt-PA different time and rt-PA group ratio
Compared with APTT, PT and TT level are without significant difference (P > 0.05), such as Fig. 7 B, C).Solvent control group and rt-PA group FIB are extremely aobvious
Write property be higher than sham-operation group (**P < 0.01), compared with solvent control group, after individually giving Gastrodin (120mg/kg), FIB is omited
There is decline.Gastrodin is administered compared with rt-PA group in rt-PA different time, and serum FIB is on close level, such as Fig. 7 D.
4.8 brain tissue MMP-9 contents
24 hours after MCAO Reperfu- sion, the extremely significant property raising of solvent control group and rt-PA group brain tissue MMP-9 content (**
P < 0.01), and after giving rt-PA, brain tissue MMP-9 has further raised trend.Gastrodin group and solvent control group ratio
Compared with brain tissue MMP-9 changes of contents is unobvious.Compared with rt-PA group, Gastrodin is administered in rt-PA different time, brain tissue
MMP-9 level is on a declining curve, sees Fig. 8.
4.9 serum inflammatory factors of senile
24 hours after MCAO Reperfu- sion, compared with sham-operation group, the horizontal extremely significant property of -1 β of solvent control group serum IL is increased
(**P < 0.01), rt-PA group serum IL -1 β is horizontal and TNF α conspicuousness increase (*P < 0.05), such as Fig. 9 A, D；With solvent control
Group is compared, Gastrodin be applied alone the horizontal conspicuousness of group IL-1 β reduce (#P < 0.05), such as Fig. 9 A, give blood serum IL-6 after rt-PA,
IL-12 and TNF α have further raising.Gastrodin is applied alone or is administered in rt-PA different time, serum IL -1 β, IL-6, IL-
12 and TNF α have decreasing trend, see Fig. 9.
4.10 general state is observed
After MCAO modeling, rat autonomic activities, which are showed, to be rotated or is tilted to left side of body, and normal walking function is by difference
The influence of degree.Medicine-feeding part has no adverse reaction.After 2 hours Reperfu- sions of ischemic, there is the animal of different number in modeling each group
Death, compared with solvent control group, the groups of animals death time for giving rt-PA is shifted to an earlier date.Rat performance is apathetic,
Activity is reduced, and feed drinking-water activity is reduced.After Reperfu- sion 24 hours, groups of animals The dead quantity be increased.
4.11 weight
24 hours after MCAO Reperfu- sion, solvent control group and the extremely significant property of rt-PA group weight loss percentage are higher than vacation
Operation group (**P < 0.01), see Figure 10.Gastrodin be applied alone or rt-PA different time be administered, weight loss trend under
Drop.
5 conclusions
This test blocks intraluminal middle cerebral artery occlusion in rats using line brush, and Reperfu- sion gives clinical agent simultaneously after ischemic 2 hours
It measures rt-PA (5mg/kg) and induces bleeding, simulate hemorrhagic conversion after cerebral apoplexy thrombolysis caused by clinically rt-PA, study Gastrodin
(120mg/kg) causes bleeding the protective effect and mechanism of conversion to rt-PA.
After ischemic Reperfu- sion 24 hours 2 hours, the rt-PA group death rate and brain tissue hemoglobin level be not obviously higher than
The solvent control group of rt-PA is given, hemorrhagic conversion after cerebral apoplexy thrombolysis caused by clinically rt-PA is successfully simulated.In this test
In, vein gives Gastrodin (120mg/kg), can reduce ischemic side cerebral infarction volume to a certain degree；The 60min before giving rt-PA
Gastrodin (120mg/kg) is given, has different degrees of relaxation effect to rt-PA induction brain tissue bleeding.
In this research, Gastrodin can reduce to a certain degree ischemic side brain tissue MMP-9 level, and reduce IL-1 β, IL-6,
The serum inflammatory parameters such as IL-12, TNF α illustrate that Gastrodin may lead ischemia-reperfusion by anti-inflammatory, inhibition oxidative stress
Brain tissue impairment and rt-PA the induction brain tissue hemorrhagic conversion of cause play protective effect.
Embodiment 2: Gastrodin is to the HBMEC protective effect of OGD induced damage and Mechanism Study
1. experimental material
1.1 test samples: Injectio of gastrodine, Kun Yao Group Plc, character: colourless transparent liquid；Specification:
2mL:0.2g/ branch；Lot number 17JX06-222；Preservation condition: room temperature is protected from light；Validity period: 2020.09；
1.2 reference substances: title or code name: nimotop vial, lot number: 180320, specification: 20mL:4mg, sub- treasured medicine
Industry.
1.3 experimental cells: Human Brain Microvascular Endothelial strain (HBMEC) is purchased from the limited public affairs of Guangzhou Ji Niou biotechnology
Department.
1.4 other reagents: Endothelial cell culture base (ECM), ScienCell；DMEM in high glucose basal medium, sugar-free DMEM
Basal medium, DPBS Du Shi phosphate buffer, PBS phosphate buffer, fetal calf serum (FBS), 0.25%Trypsin-EDTA
(1X) is purchased from Gibco；Blueness-streptomysin (dual anti-), BI；Dimethyl sulfoxide (DMSO), Sigma；CellTiterAQueous
One Solution Cell Proliferation Assay (MTS), Promega；PrimeScriptTMRT reagent Kit
With gDNA Eraser (Perfect Real Time), TB GreenTMPremix Ex TaqTMII(Tli RNaseH
Plus), TaKaRa；Protein Extraction Reagent kit, BestBio；BCA method measures protein concentration, Boster；5 × albumen loading buffer
Liquid (contains DTT), 5%BSA confining liquid, 20 × TBST buffer, 5 × Tris- glycine running buffer, and 10 × electrophoretic transfer is slow
Fliud flushing, Solarbio；TGXTMFastCastTMAcrylamide Kit, BIO-RAD；TRNzol A+Total RNA extraction reagent box, mistake
Ammonium sulfate (AP), Aladdin；RNase-Free ddH2O, TIENGEN；DEPC water, ECL chemiluminescent substance, Biosharp；
Chloroform (AR), isopropanol (AR), dehydrated alcohol (AR), anhydrous methanol (AR) are domestic.
Main primer:
| Gene Name | Upstream primer sequence | Downstream primer sequence |
| Claudin-5 | GAGGCGTGCTCTACCTGTTT | AGCTCGTACTTCTGCGACAC |
| Occludin | ACTTCAGGCAGCCTCGTTAC | CCTGATCCAGTCCTCCTCCA |
| ZO-1 | AGCCATTCCCGAAGGAGTTG | ATCACAGTGTGGTAAGCGCA |
| GAPDH (internal reference) | GAGAAGGCTGGGGCTCATTT | AGTGATGGCATGGACTGTGG |
Note: experiment the primer is humanized
Main antibody:
| Antibody | Extension rate | Manufacturer |
| Claudin5 antibody (rabbit-anti) | 1:1000 | Sigma |
| Occludin antibody (rabbit-anti) | 1:1000 | Sigma |
| ZO-1 antibody (rabbit-anti) | 1:1000 | Cell Signaling Technology |
| GAPDH antibody (rabbit-anti) | 1:1000 | Cell Signaling Technology |
| Goat-anti rabbit secondary antibody | 1:10000 | Sigma |
Note: testing antibody used is humanized
1.5 experimental facilities and consumptive material: superclean bench, SERIES II WATER JACKET carbon dioxide incubator surpass
Low temperature refrigerator, Biofuge high-speed refrigerated centrifuge, Thermo Forma company, the U.S.；XD-202 inverted microscope, Nanjing Jiangnan
Optical Co., Ltd, Yongxin；The U.S. MIC101 billups-rothenberg cell hypoxia research device, U.S. Chauncey Billups sieve
Sen Bao company；BioTek Synergy HT multi-function microplate reader, BioTek Instruments company, the U.S.；StepOne
Plus real-time fluorescence quantitative PCR instrument, VeritiTM PCR instrument, Applied Biosystems company, the U.S.；Vertical electrophoresis-transferring film
System, Western blot transferring film filter paper, BIO-RAD company, the U.S.；ELITE 300PLUS electrophoresis apparatus, U.S. Wealtec are public
Department；GelDoc-It2 gel imager, UVP company, the U.S.；STS-3 decolorization swinging table, Shanghai Qi Te Analytical Instrument Co., Ltd；
XW-80A vortex mixer, Instrument Factory of Shanghai Medical Univ.；Haier's Medical refrigerator, biologic medical limited liability company, Haier；HH-
420-2B electric heat constant temperature water tank, Shanghai Zuo Le Instrument Ltd.；METTLER-TOLEDO MS semimicro-analy-tical balance, Switzerland Mei Te
Le-support benefit company；Program temperature reduction box, Nalgene company, the U.S.；MiniSpin centrifuge, micro sample adding appliance, manual liquid relief
Rifle, German Eppendorf company；Milli-Q Reference ultrapure water machine, pvdf membrane, Millipore company, the U.S.；Snow section
Ice machine, Xue Ke Electrical Appliances Co., Ltd, Changshu City；Tissue Culture Dish, centrifuge tube, EP pipe, tissue culture plate, ELISA Plate, the U.S.
Corning company.
2. experiment content
2.1 cell culture: being removed from liquid nitrogen the HBMEC cell frozen, and the fresh ECM culture medium of people is added to train
It supports, is placed in containing 5%CO237 DEG C of incubators, 2-3 days cells change liquid 1 time.It is thin up to 90% or more, DPBS cleaning to cell density
Born of the same parents, pancreatin digestion after cell is resuspended with fresh culture, passes 2 ratio according to 1, cell suspension are added separately to different
In culture dish, addition culture medium continues to cultivate.
The investigation of 2.2 OGD induction HBMEC damage model modeling conditions: experiment uses the U.S. MIC101 billups-
Rothenberg cell hypoxia research device provides anaerobic environment, i.e. Anoxia model (Oxygen glucose for cell
deprivation,OGD).Determination of probing into according to laboratory early period to HBMEC growth cycle and growth curve, HBMEC with 1 ×
104A/hole cell density is inoculated in 96 orifice plates, and when 36h enters cell log growth period, and cell density is up to 90% or so when 60h.
Multi-function microplate reader detects the absorbance value of cell at 490nm, and computation model group cell OGD handles different time, and cell is living
The variation of power.Calculation formula are as follows:(blank
Control group is to contain only ECM culture medium and detection reagent)
HBMEC protective effect of 2.3 Injectio of gastrodine to OGD induced damage
2.3.1 inoculating cell: by HBMEC cell inoculation in 96 porocyte culture plates, every group of 6 multiple holes, every 100 μ L of hole is thin
Born of the same parents' suspension, covers plate lid, after label, 37 DEG C, and 5%CO2Culture is incubated in incubator.
2.3.2 experimental group: being divided into normal group, model group, Nimodipine group and Gastrodin group, every group and each drug it is dense
Degree group is 6 multiple holes.After kind plate culture 36h, culture plate is taken out, slowly exhausts former culture medium, normal group and mould in cell hole
Type group cell hole is directly added into fresh ECM culture medium, and medicine group is separately added into containing final concentration of nimotop vial stoste
1/40,1/100,1/500, the 1/2500 of 1/200,1/1000 (11.950,2.390,0.478 μM) and Injectio of gastrodine stoste
The fresh ECM culture medium of (3.386,0.677,0.135mM), 100 μ L of every hole, 37 DEG C, 5%CO2Continue to cultivate in incubator.
2.3.3 OGD MODEL DAMAGE: after dosing culture for 24 hours (i.e. kind plate 60h), the DPBS after being preheated with 37 DEG C of water-baths embathes
Twice, every hole 100 μ L every time exhausts DPBS to cell, and fresh ECM culture medium is added in normal cell hole of organizing, and is put into 37 DEG C, and 5%
CO2Continue to cultivate in incubator.Sugar-free DMEM culture solution is added in model group cell hole.Medicine group is added with drug containing final concentration
Sugar-free DMEM culture solution, every 100 μ L of hole, model group and medicine group are put into together and are pre-filled with 95%N2+ 5%CO2Gaseous mixture
In the hypoxia device of body, then mixed gas 2-4min is continually fed into 20L/min flow velocity, guarantees O in device2Content is lower than 2%,
Gas exchanges terminate, and seal hypoxia device, alcohol disinfecting, put it into 37 DEG C of incubators continue to cultivate respectively cell 1,3,4,
9h。
2.3.4 MTS method detects cell viability: after cell culture to stipulated time, taking out cell respectively in multifunctional enzyme mark
The absorbance value that cell is measured under instrument 490nm calculates nimotop vial and each concentration group of Injectio of gastrodine and damages to OGD
Under HBMEC vigor influence.Calculation formula are as follows: (blank control group is to contain only ECM culture medium
With detection reagent)
Influence of 2.4 Injectio of gastrodine to HBMECClaudin-5, Occludin and ZO-1 of OGD induced damage
2.4.1 qPCR detects Injectio of gastrodine to OGD induced damage HBMEC Claudin-5, Occludin and ZO-1
The influence of mRNA transcriptional level
After HBMEC cell culture to stipulated time, Total RNAs extraction is carried out, and be measured with purity to its concentration.It presses
PrimeScriptTMRT reagent Kit with gDNA Eraser kit specification removes gRNA, and says by kit
Bright book, by following ingredient in preparing reaction mixture on ice, in the reaction tube after then dispensing 10 μ L to each removal gDNA again.
Reverse transcription reaction is carried out by such as following table 2-1 program immediately after soft mixing.
Table 2-1 RNA reverse transcription cDNA reaction system
| Reagent | Usage amount |
| Reaction solution after removing gDNA | 10μL |
| PrimeScript RT Enzyme Mix I | 1μL |
| RT Primer Mix*4 | 1μL |
| 5×PrimeScript Buffer 2(for Real Time) | 4μL |
| RNase Free dH2O | 4μL |
| Total | 20μL |
3 multiple holes are arranged in each gene of each sample, and the negative control of each gene is also provided with 3 multiple holes, with GAPDH
For internal reference, by TB GreenTMPremix Ex TaqTMII kit specification, will be if following table 2-2 ingredient is in preparation reaction on ice
Mixed liquor after soft mixing, is carried out by setting experimental arrangement, linear regression equation is obtained by standard curve, according to 2-△△CtIt calculates
Method obtains target gene mRNA relative expression quantity.
Table 2-2 polymerase chain reaction system
| Reagent | Usage amount | Total concentration |
| TB Green Premix Ex Taq II(2×) | 10μL | 1× |
| PCR Forward Primer(10μM) | 0.8μL | 0.4μM |
| PCR Reverse Primer(10μM) | 0.8μL | 0.4μM |
| ROX Reference Dye(50×) | 0.4μL | 1× |
| DNA profiling | 2 μ L (μ of <=2 L) | — |
| Aqua sterilisa | 6 μ L (are mended to 20 μ L) | — |
| Total | 20μL | — |
2.4.2 Westernblot detects Injectio of gastrodine to OGD induced damage HBMECClaudin-5, Occludin
With the influence of ZO-1 protein expression level
Illustrate to prepare protein extract (inner protein Protease Inhibitor Cocktail and phosphatase inhibition by Protein Extraction Reagent kit
Agent composition), measurement adjustment protein concentration, by the way that denaturation, glue, loading, electrophoresis, revolving die, closing, primary antibody are incubated for, secondary antibody is incubated
Educate, develop gel image analysis, take pictures, using Image J software to result carry out gray analysis, the relative expression of destination protein
Amount=destination protein gray value/internal reference albumen gray value.
2.6 statistical analysis
For statistical analysis with 23.0 statistical software of IBM SPSS, each group experimental data is all made of " mean ± standard
Difference "It indicates, multiple groups sample room mean compares using one-way analysis of variance (one-wayANOVA), compares adopt two-by-two
It is examined with LSD.With α=0.05 be inspection level P < 0.05 when difference it is statistically significant.
3. test result
HBMEC protective effect of 3.1 Injectio of gastrodine to OGD induced damage
3.1.1 OGD induces the investigation of HBMEC damage model modeling conditions
As shown in table 2-3 and Figure 11, HBMEC density is incubated for 9h, MTS method measures up to after 90% under Anoxia state
Cell survival rate be reduced to 50% or so, cellular damage degree is moderate at this time.And after cell OGD damage 12h, cell survival rate is not
To 40%, cell has formed irreversible damage at this time, is unsuitable for later experiments research.Therefore use OGD damage 1,3,6,9h for
The modeling time, to investigate influence of the drug to cell survival rate.
Table 2-3 OGD induce HBMEC damage different time after cell survival rate (N=6)
Compared with other times:*P<0.01
3.1.2 HBMEC protective effect of the Injectio of gastrodine to OGD induced damage
As shown in Table 2-4, HBMEC density is incubated for 1h, each group is poor without statistics up to after 90% under Anoxia state
It is different；Cell OGD damages 3h, and model group cell survival rate is substantially less than normal group, nimotop vial stoste dilution 200,
Three concentration groups of 1000 times of groups and Injectio of gastrodine play the role of protecting cell compared with model group；Cell OGD damage
Hurt 6h, nimotop vial stoste dilution three concentration groups of 1000 times of groups and Injectio of gastrodine can still reduce Anoxia pair
The degree of injury of cell；Cell OGD damages 9h, and each medicine group does not play cytoprotection.
As shown in table 2-4 and Figure 12, by comparing " medicine group cell survival rate (%)-OGD organizes cell survival rate (%) "
As a result, can when cell hypoxia lacks sugar 3h, Injectio of gastrodine stoste 500 times of administration groups of dilution to the protective effect of cell most
By force, at this point, nimotop vial stoste dilutes the cell survival rate of 1000 times of groups compared with other two concentration group height.Therefore, originally
Experimental selection OGD MODEL DAMAGE 3h probes into the modeling condition of mechanism of drug action as the later period, and nimotop vial stoste is dilute
1000 times are released, Injectio of gastrodine stoste dilutes 500 times as later experiments drug and uses concentration.
The above experimental result prompt, when cell hypoxia sugar deficiency injury degree is slight and serious, drug fails to embody damage-retardation
The effect of wound, especially high concentration medicine.But cell has damaged, and when belonging to early stage reversible lesion, suitable concentration
Drug shows stronger cytoprotection.Therefore, when oxygen-glucose deprived injury occurs, the medicine of suitable concentration should be used as early as possible
Object reduces cellular damage, improves survival rate.
Shadow of the Injectio of gastrodine of table 2-5 various concentration to cell survival rate after OGD induction HBMEC damage different time
Ring (N=6)
Influence of 3.2 Injectio of gastrodine to OGD induced damage HBMEC Claudin-5, Occludin and ZO-1
3.2.1 Injectio of gastrodine transcribes water to OGD induced damage HBMECClaudin-5, Occludin and ZO-1mRNA
Flat influence
As shown in table 2-6 and Figure 13 result, HBMEC is incubated for 3h under Anoxia state, model group Claudin-5,
The relatively normal group of Occludin and ZO-1 mRNA transcriptional level is significant to increase (P < 0.01), three bases of nimotop vial group
Because transcriptional level significantly increases (P < 0.01) compared with model group, Injectio of gastrodine group also significantly raises Claudin-5 compared with model group
With the transcriptional level (P < 0.01) of OccludinmRNA.In the case where oxygen-glucose deprived injury, cell can raise itself for prompt
Claudin-5, Occludin and ZO-1 mRNA transcriptional level, while Injectio of gastrodine is to Claudin-5 and Occludin
MRNA also has significant up-regulation to act on.
Table 2-6 group of cells related gene mRNA transcription situation (N=3)
Compared with normal group:*P < 0.05,**P<0.01；Compared with model group:#P < 0.05,##P<0.01；
N: normal group；M: model group；Nimodipine: nimotop vial stoste dilutes 1000 times；Gastrodin:
Injectio of gastrodine stoste dilutes 500 times.Compared with normal group:**P<0.01；Compared with model group:##P<0.01；
3..2.2 Injectio of gastrodine is to OGD induced damage HBMEC Claudin-5, Occludin and ZO-1 protein expression
Horizontal influence
As shown in table 2-7 and Figure 14, Figure 15, HBMEC is incubated for 3h, model group Occludin closure under oxygen-glucose deprived injury
Protein level more normally organizes reduction (P < 0.05)；Compared with model group: nimotop vial group be remarkably improved Claudin-5 and
The expression of Occludin albumen, and have the tendency that raising the band-like Occludin expression of ZO-1, but without statistically indifference；
Injectio of gastrodine also can significantly raise the effect of Claudin-5 and Occludin protein expression level, but to ZO-1 albumen without
It influences.Prompt Injectio of gastrodine that there is certain promotion to Claudin-5 and Occludin protein expression.
Table 2-7 group of cells Claudin-5, Occludin and ZO-1 protein expression situation (N=3)
Compared with normal group:*P<0.05；Compared with model group:#P<0.05；
The present invention detects day with qPCR and Western blot technology by external OGD induced damage single layer HBMEC
Numb element injection is transcribed to intracellular Claudin-5, Occludin and ZO-1mRNA and the influence of protein expression level, prompts
OGD induces HBMEC damage early stage, and Injectio of gastrodine, which mainly passes through, adjusts Claudin-5 and Occludin mRNA transcription and egg
White expression, to maintain TJs structure and function stability.
Research institute obtains as a result, oxygen-glucose deprived injury is prompted to give the Gastrodin of suitable concentration in advance in early days inside and outside combination
Injection has certain protective effect to TJs and BBB.It is treated clinical use plasminogen activator (rt-PA) and lacks
Hemorrhagic brain soldier and existing hemorrhagic conversion can also play certain inhibiting effect, the opposite thromboembolism treatment window for extending rt-PA,
Strive for more treatment times for clinical patients with cerebral apoplexy, the injury of disease Central nervous system is further decreased, after making more
Sequelae is well reduced, patients ' life quality is improved.
The above is only presently preferred embodiments of the present invention, is not intended to limit the present invention in any form, though
So the present invention has been disclosed as a preferred embodiment, and however, it is not intended to limit the invention, any to be familiar with technology people of the invention
Member without departing from the scope of the present invention, when the technology contents using above-mentioned prompt make it is a little variation or be modified to
The equivalent embodiment of equivalent variations, but anything that does not depart from the technical scheme of the invention content, it is right according to the technical essence of the invention
Any simple modification, equivalent change and modification made by above embodiments, in the range of still falling within the present invention program.
Claims (10)
1. application of the Gastrodin in preparation treatment cerebral hemorrhage drug, which is characterized in that the cerebral hemorrhage is cerebral ischemia rt-PA
Caused hemorrhagic conversion after thromboembolism treatment.
2. application according to claim 1, which is characterized in that go out caused by after the cerebral ischemia rt-PA thromboembolism treatment
Blood is converted into hemorrhagic conversion caused by MMP-9 increased activity after cerebral ischemia rt-PA thromboembolism treatment.
3. application according to claim 1 or 2, which is characterized in that caused by after the cerebral ischemia rt-PA thromboembolism treatment
Hemorrhagic conversion is IL-1 β, IL-6 and/or the caused hemorrhagic conversion of IL-12 overexpression after cerebral ischemia rt-PA thromboembolism treatment.
4. application according to claim 1 to 3, which is characterized in that the cerebral ischemia rt-PA thromboembolism treatment
Caused hemorrhagic conversion is the caused hemorrhagic conversion of TNF α expression enhancing after cerebral ischemia rt-PA thromboembolism treatment afterwards.
5. application according to any one of claims 1-4, which is characterized in that the cerebral ischemia rt-PA thromboembolism treatment
Caused hemorrhagic conversion is the caused hemorrhagic conversion of TJs correlative protein expression level decline after cerebral ischemia rt-PA thromboembolism treatment afterwards.
6. Gastrodin is preparing the application in MMP-9 activity inhibitor drug or TNF α expression inhibiting agent drug.
7. Gastrodin prepare IL-1 β, the application in IL-6, IL-12 expression inhibiting agent drug.
8. application described in -7 any one according to claim 1, which is characterized in that the drug further includes that can pharmaceutically connect
The auxiliary material received；It is preferred that the dosage form of the drug is injection, tablet, capsule, pulvis, pill or oral solution.
9. a kind of combination product containing the first medicament and second medicament, which is characterized in that first medicament is rt-PA, described
Second medicament is Gastrodin.
10. application of the composition product as claimed in claim 9 in preparation treatment acute ischemic cerebral apoplexy drug.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111150737A (en) * | 2020-01-17 | 2020-05-15 | 昆明医科大学 | New application of gastrodin |
| CN117838803A (en) * | 2024-02-01 | 2024-04-09 | 浙江大学 | Application of gastrodia elata-derived nano extracellular vesicles in preparation of drugs for preventing and/or treating subarachnoid hemorrhage |
-
2019
- 2019-05-20 CN CN201910418762.6A patent/CN110179808A/en active Pending
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| SHAOQING WANG等: "Gastrodin improves the neurological score in MCAO rats by inhibiting inflammation and apoptosis, promoting revascularization", 《INT J CLIN EXP PATHOL》 * |
| SHIPENG LI等: "Gastrodin pretreatment alleviates rat brain injury caused by cerebral ischemic-reperfusion", 《BRAIN RESEARCH》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111150737A (en) * | 2020-01-17 | 2020-05-15 | 昆明医科大学 | New application of gastrodin |
| CN117838803A (en) * | 2024-02-01 | 2024-04-09 | 浙江大学 | Application of gastrodia elata-derived nano extracellular vesicles in preparation of drugs for preventing and/or treating subarachnoid hemorrhage |
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