CN105998276A - Application of total flavones of oldenlandia diffusa willd in preparing drugs for treating meningeal microcirculation disturbance and cerebral ischemia by serving as only active site - Google Patents
Application of total flavones of oldenlandia diffusa willd in preparing drugs for treating meningeal microcirculation disturbance and cerebral ischemia by serving as only active site Download PDFInfo
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Abstract
The invention relates to application of total flavones of oldenlandia diffusa willd in preparing drugs for treating meningeal microcirculation disturbance and cerebral ischemia by serving as the only active site. The total flavones of oldenlandia diffusa willd are obtained through the steps that oldenlandia diffusa willd is smashed into crude powder, reflux extraction is conducted with petroleum ether, and degreasing is conducted; residues with petroleum ether volatilized completely are soaked with ethyl alcohol, reflux extraction is conducted twice, filtrate for the two times are combined, ethyl alcohol is subjected to vacuum recycling till no alcohol taste exists, distilled water is added, the treated filtrated is dispersed into a sample solution, AB-8 type macroporous adsorption resin is applied to the sample solution, elution is conducted with the distilled water, and water liquid is discarded; elution is conducted with 10% ethyl alcohol and then with 80% ethyl alcohol, 80% ethyl alcohol eluant is collected, ethyl alcohol is subjected to vacuum recycling till no alcohol taste exists, freeze drying and smashing are conducted, and the total flavones of oldenlandia diffusa willd are obtained. According to the application of the total flavones of oldenlandia diffusa willd in preparing the drugs for treating the meningeal microcirculation disturbance and cerebral ischemia by serving as the only active site, the effect of improving the meningeal microcirculation disturbance and cerebral ischemia is achieved, the total flavones of oldenlandia diffusa willd serving as the only active site are effectively used for preparing the drugs for treating the meningeal microcirculation disturbance and cerebral ischemia, and innovation on treating the meningeal microcirculation disturbance and cerebral ischemia is achieved.
Description
Technical field
The present invention relates to medicine, particularly total flavones of oldenlandia diffusa willd is sole active position meninges is micro-follows in preparation treatment
Application in ring obstacle, brain ischemia medicament.
Background technology
Microcirculation, directly to cell blood supply, oxygen supply, energizing quantity and relevant nutrient substance, is the most also discharged harmful
Metabolite, be the interior environment of human body, be the most basic guarantee of life.Any organ of human body, any position (includes the heart
Including dirty) all it is necessary to have the microcirculation of a normal health, otherwise arise that the pathological changes of corresponding organ.
Under normal circumstances, microcirculatory blood flow adapts with tissue, organ metaboilic level, makes each organ of inside of human body
Physiological function is normally carried out.If microcirculation is obstructed, just looking like the water channel blocking in one piece of rice seedling bed, the seedlings of cereal crops just can not get moisture
As meeting is withered, also can there is disease and aging etc. because metabolism is abnormal in human internal organ.When barrier occurs in meninges microcirculation
When hindering, arise that the symptoms such as transient ischemic attack.
Transient ischemic attack (TIA) is carotid artery or Vertebro-basilar System generation transience blood supply deficiency,
Focal cerebral ischemia is caused to cause burst, transience, reversibility delayed ischemic neurological deficits.Outbreak last for several minutes, generally 30
Recover completely in minute, little more than 2 often leave over slight neurologic impairment performance, or CT and MRI display brain tissue ischemia levy
As, male is more than women.How morbidity suddenly, is sent out when Body Position Change, hyperkinesia, cervical region rotate suddenly or bend and stretch
Sick.Morbidity absence of aura, has transient focal neurological signs, general unconscious obstacle, lasts 5～20 minutes, can be repeatedly
Outbreak, but typically recovered completely in 24 hours, no sequel, but to the healthy harmful effect having in various degree.Existing
Though having treatment meninges microcirculation, the multi-medicament of cerebral ischemia, but for various reasons, its use and curative effect are the most unsatisfactory, because of
This, develop and develop new medicine imperative.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the purpose of the present invention is just to provide a kind of Herba Hedyotidis Diffusae
Total flavones is the application in preparation treatment meninges microcirculation disturbance, brain ischemia medicament of the sole active position, can effectively solve white
The preparation of Serpentis SHECAO total flavones, and the application problem in preparation treatment meninges microcirculation disturbance, brain ischemia medicament.
The technical scheme that the present invention solves is, a kind of total flavones of oldenlandia diffusa willd is that sole active position is at preparation treatment brain
Application in film microcirculation disturbance, brain ischemia medicament, described total flavones of oldenlandia diffusa willd is to be ground into slightly by Herba Hedyotidis Diffusae
Powder, first petroleum ether reflux, extract, defat；Medicinal residues volatilize petroleum ether, with soak with ethanol, reflux, extract, 2 times, merging filtrate, decompression
Recovery ethanol, to without alcohol taste, adds distilled water and is dispersed into sample liquid, upper AB-8 type macroporous adsorbent resin, first distilled water eluting, discard
Water liquid；Use 10% ethanol elution, then use 80% ethanol elution, collect 80% ethanol elution, decompression recycling ethanol to without alcohol taste,
Lyophilization, pulverizes, obtains total flavones of oldenlandia diffusa willd.
Total flavones of the present invention is the active site extracted from Herba Hedyotidis Diffusae, has and improves meninges microcirculation disturbance, brain
Effect of ischemia, is effective to preparation treatment meninges microcirculation disturbance, brain ischemia medicament as sole active position, has opened up white
The new application of Serpentis SHECAO, is the innovation on treatment meninges microcirculation disturbance, brain ischemia medicament, has the most economic and society's effect
Benefit.
Detailed description of the invention
Below in conjunction with concrete condition, the detailed description of the invention of the present invention is elaborated.
A kind of total flavones of oldenlandia diffusa willd is that sole active position is at preparation treatment meninges microcirculation disturbance, brain ischemia medicament
In application, described total flavones of oldenlandia diffusa willd is, Herba Hedyotidis Diffusae was ground into the coarse powder of 30 mesh sieves, first adds coarse powder 10
Times weight petroleum ether reflux, extract, 1h, defat, discard petroleum ether, obtain medicinal residues；Medicinal residues volatilize petroleum ether, add every time 10 times of weight,
Mass concentration 80% soak with ethanol 0.5h, reflux, extract, 2 times, each 1h, filter, merging filtrate, decompression recycling ethanol is to without alcohol
Taste, adds distilled water and is dispersed into and is equivalent to, containing crude drug amount 0.3g/ml, obtain sample liquid；AB-8 type macroporous adsorbent resin in sample liquid, first
With 2 times of column volume distilled water eluting, discard water liquid；Continue with 4 times of column volumes, mass concentration 10% ethanol elution, discard eluent；
Again with 6 times of column volumes, mass concentration 80% ethanol elution, collecting 80% ethanol elution, decompression recycling ethanol is to without alcohol taste, cold
Freeze and dry, pulverize, obtain total flavones of oldenlandia diffusa willd.
Total flavones of oldenlandia diffusa willd prepared by the present invention, as sole active position, has and improves meninges microcirculation, anti-brain
Effect of ischemia, and obtained fully proving through experiment, relevant testing data is as follows:
Test total flavones of oldenlandia diffusa willd impact microcirculatory on mice meninges
1 experiment material
1.1 Experimental agents
Total flavones of oldenlandia diffusa willd, is provided by Henan College Of Traditional Chinese Medicine chemistry room, and content is 62%；Nimodipine tablet, sub-precious
Pharmaceutical Group Plc produces (every contains 20mg)；NAOLUOTONG JIAONANG, Jilin Jinbao Pharmaceutical Co., Ltd. produces
(every 0.5g).
1.2 instrument
Speckle full frame real time scan imaging system, PIM3 manufacturer: handkerchief auspicious medical science and technology (Beijing) company limited.
1.4 laboratory animal
KM mice 70, SPF, male and female half and half, body weight 18-22g, purchased from Henan Province's Experimental Animal Center, this batch of mice is closed
Lattice card number: NO.41003100001113.
2 experimental techniques
2.1 modelings and administration
Taking 18-22g healthy mice 70, male and female half and half, weigh, be divided at random: blank group, model group, brain network leads to group,
Nimodipine group, total flavones of oldenlandia diffusa willd large, medium and small dosage group, totally 7 groups, often group 12.In addition to blank group, make for remaining 6 groups
Mice bilateral ligation model.Nimodipine group (positive drug, is made into the suspension of 3mg/ml by 0.1ml/10g,
30mg/kg, 15 times of quantity)；NAOLUOTONG JIAONANG group (positive drug, is made into the suspension of 75mg/ml by 0.1ml/10g,
750mg/kg, 15 times of quantity)；Total flavones of oldenlandia diffusa willd large, medium and small dosage group (is made into 40mg/ by 0.1ml/10g
The suspension of ml, 20mg/ml, 10mg/ml, 400mg/kg, 200mg/kg, 100mg/kg, 15 times of quantity)；Preoperative, give
Give each corresponding group mice nimodipine, brain network leads to, the suspension of the large, medium and small dosage of total flavones of oldenlandia diffusa willd gavages, blank
Group, model group only gavage commensurability sodium carboxymethyl cellulose (0.5%CMC), 1 1d, serve on 7d.Press 6d evening 8 and batch prohibit
Water be can't help by food, after 7d 8 a.m. is by criticizing the administration 1h that weighs, starts test.First computer and other peripheral device are started, so
Rear startup Perimed instrument, preheats 5 minutes, can start monitoring when the LED of instrument stops flicker, double-click on desktop
PIMsoft software, selects PeriCam PSI system.Click tools, arranging operating distance is 10cm, the width of monitored area and
Being highly 2cm, during step-length is set to, frequency is set to 23/s.Mice is with 5% chloral hydrate 0.06ml/10g intraperitoneal injection of anesthesia
(righting reflex loss of mice), ventricumbent position is fixed, and skull hits exactly skin sagittal and cuts, drawn by both sides skin with mosquito forceps
Open, make a labelling with marker pen at mouse brain sagittal suture and coronal suture infall, mice is placed in the recommended distance model of instrument
In enclosing, and making instrument parallel with monitoring position, light beam impinges upon mark, and vertical with detection position, adjusts instrument distance and sets
Set a distance is consistent, and delineation scan area is clicked on record button and started record, and record mice is blood perfusion in tranquil latter 2 minutes
Amount.Unclamping mice, dorsal position is fixed, every side common carotid artery occlusion (blank group i.e. sham operated rats only separates bilateral common carotid arteries,
Do not do ligation to process), then to unclamp mice, and ventricumbent position is fixed, record mice is blood perfusion amount in tranquil latter 2 minutes.
2.2 Testing index
Take cut prick front 1 point 50 seconds--the blood perfusion amount meansigma methods of 2 minutes as cut prick before average irrigation amount, take 3 points
50 seconds--the blood perfusion amount meansigma methods of 4 minutes, as the average irrigation amount cut after pricking, deducts by the blood perfusion amount after ligation
Blood perfusion amount before ligation measures out the rate of descent of cerebral blood flow divided by the blood perfusion before ligation.
2.2.1 assay method
In addition to blank group, remaining mice is all cooked Bilateral Cervical and always ligatures, and then starts examination after 1 hour in preoperative administered in portions
Test, start computer and other peripheral device, then start Perimed instrument, preheat 5 minutes, when the LED of instrument stops dodging
Monitoring can be started time bright, double-click the PIMsoft software on desktop, select PeriCam PSI system.Click tools, arranges behaviour
Making distance for 10cm, the width of monitored area and height are 2cm, and during step-length is set to, frequency is set to 23/s.Mice 5% water
Closing chloral 0.06ml/10g intraperitoneal injection of anesthesia (righting reflex loss of mice), ventricumbent position is fixed, and skull hits exactly skin and vows
Shape cuts, and is pulled open by both sides skin with mosquito forceps, makees a labelling with marker pen at mouse brain sagittal suture and coronal suture infall,
In the range of mice is placed in the recommended distance of instrument, and making instrument parallel with monitoring position, light beam impinges upon mark, and with detection
Position is vertical, adjusts instrument distance consistent with setpoint distance, and delineation scan area is clicked on record button and started record, records little
Mus is blood perfusion amount in tranquil latter 2 minutes.Unclamp mice, and dorsal position is fixed, (the blank group i.e. vacation of every side common carotid artery occlusion
Operation group only separates bilateral common carotid arteries, does not do ligation and processes), then to unclamp mice, and ventricumbent position is fixed, record mice is tranquil
Blood perfusion amount in latter 2 minutes, derives report, is analyzed.
3. statistical procedures method
Data analysis SPSS 17.0 statistical package carries out the statistical procedures of data information, measurement data: average
± standard deviationComparing between each group: one factor analysis of variance, variance is together with least significant difference (LSD) method, heterogeneity of variance
Check by Games-Howell method.
4 experimental results
The impact microcirculatory on mice meninges of 4.1 total flavones of oldenlandia diffusa willds
The impact microcirculatory on mice meninges of table 1 total flavones of oldenlandia diffusa willd
Note: compare model group, medication group has significant difference (* * P < 0.01), hence it is evident that sex differernce (* P < 0.05)
As shown in Table 1: it is inconspicuous that neck respectively organizes mice meninges microcirculatory blood flow difference before always ligaturing, illustrate that packet is uniformly.
With sham operated rats ratio, model group meninges microcirculatory blood flow substantially reduces (P < 0.01), and modeling success is described, but because of collateral circulation
And local blood circulation, though ligation bilateral common carotid arteries, cerebral blood flow blocks the most completely；With model group ratio, large, medium and small dosage
Total flavones of oldenlandia diffusa willd group, nimodipine group and brain network lead to group all can significantly improve the mice caused by bilateral ligation
Cerebral blood flow reduces (P < 0.01)；Always ligature front ratio with neck, after common carotid artery occlusion, each group mice meninges microcirculatory blood flow is equal
Substantially reduce (P < 0.01).
5 conclusions
Always ligaturing before and after's cerebral blood flow change in two minutes by observing mouse carotid, model group cerebral blood flow is remarkably decreased,
Nimodipine group, brain network lead to group, total flavones of oldenlandia diffusa willd large, medium and small dosage group all can significantly improve mouse brain blood flow
Reduce, the results show total flavones of oldenlandia diffusa willd each administration group Bilateral Cervical can be promoted always to ligature after mice Doppler flow mapping drawn game
Portion's blood circulation, can significantly improve mouse brain microcirculation disturbance and brain circular blood stream state, and then cause after improving acute ischemia
Brain tissue impairment.Optimal with total flavones of oldenlandia diffusa willd heavy dose group.
Test the impact on mice repeated cerebral ischemia-reperfusion model of two total flavones of oldenlandia diffusa willds
1 experiment material
1.1 Experimental agents
Total flavones of oldenlandia diffusa willd, is provided by Henan College Of Traditional Chinese Medicine chemistry room, and general flavone content is 62%；Nimodipine
Sheet, Shanxi Yabao Pharmaceutical Group Corp. produces；NAOLUOTONG JIAONANG, Jilin Jinbao Pharmaceutical Co., Ltd. produces.
1.2 experiment reagent
Sodium carboxymethyl cellulose, Tianjin Heng Xing chemical reagent company limited produces；LDH test kit, producer: Nanjing is built up
Bioengineering Research Institute produces；Penicillin, North China Pharmaceutical Group Company Ltd produces；Chloral hydrate, Tianjin causes remoteization
Learn reagent company limited to produce；Normal saline, Cologne, Henan pharmaceutcal corporation, Ltd produces；Formalin, analytical pure, Yantai City is double
Double Chemical Co., Ltd.s produce；Medical ethanol, big preparation company limited of Xinxiang City three produces；MDA test kit, biology is built up in Nanjing
Graduate School of Engineering produces；NSE test kit, Calvin bio tech ltd, Jiangsu produces；Coomassie brilliant blue measures test kit,
Nanjing is built up Bioengineering Research Institute and is produced；LD test kit, Bioengineering Research Institute is built up in Nanjing；SOD test kit, Nanjing is built up
Bioengineering Research Institute；S-100 β test kit, Calvin bio tech ltd, Jiangsu；ATP test kit, biology is built up in Nanjing
Graduate School of Engineering produces.
1.3 instrument
Ultraviolet-uisible spectrophotometer, Shanghai Techcomp Instrument Ltd., model UV1000；Electronic balance, Shanghai
Precision scientific instrument company limited, model FA2204B；Adjustable pipette, Shanghai Lei Bo Analytical Instrument Co., Ltd；Ultrasound wave
Washer, Shanghai High Kudos Science Instrument Co., Ltd.；Glass homogenizer, Zhengzhou Glassware Factory；Table-type low-speed autobalance from
Scheming, is provided by Changsha Xiang Zhi centrifuge instrument company, model TDL-40B；Microplate reader, BIO-RAD company of the U.S., model
BIORAD-680。
1.4 laboratory animal
KM kind mice 105, SPF, male, body weight 28-31g, Henan Province's Experimental Animal Center provide, this batch of mice is closed
Lattice card number: 41003100001167.
2 experimental techniques
2.1 modelings and administration
Take healthy mice 105, male, body weight 28-31g, weigh, divide equally 7 groups at random, often group 15, blank group model
Group, brain network lead to group, nimodipine group, total flavones of oldenlandia diffusa willd large, medium and small dosage group.In addition to sham operated rats, remaining 6 groups equal
Make mice repeated cerebral ischemia-reperfusion model.Nimodipine group (positive drug, is made into the suspension of 3mg/ml by 0.1ml/10g,
30mg/kg, 15 times of quantity)；NAOLUOTONG JIAONANG group (positive drug, is made into the suspension of 75mg/ml by 0.1ml/10g,
750mg/kg, 15 times of quantity)；Total flavones of oldenlandia diffusa willd large, medium and small dosage group (is made into 40mg/ by 0.1ml/10g
The suspension of ml, 20mg/ml, 10mg/ml, 400mg/kg, 200mg/kg, 100mg/kg, 15 times of quantity)；Preoperative, give
Give each corresponding group mice nimodipine, brain network leads to, the suspension of the large, medium and small dosage of total flavones of oldenlandia diffusa willd gavages, blank
Group, model group only gavage commensurability 0.5%CMC, 1 1d, serve on 7d.
At 8 in 6d evening, water is can't help in fasting in batches, after 7d 8 a.m. batch weighing is administered 1h, by 0.03ml/10g
Ip10% chloral hydrate (righting reflex loss of mice), sterilizes with medical ethanol cervical region, then cuts cervical region center with scalpel
Position to the left, separates Surrounding muscles, peels off side nerve with elbow precision tweezer, separates CCA, every side common carotid artery acupuncture needle
Hook (diameter 0.3mm), angle about 90, ischemia 10min, perfusion 10min, then ischemia 10min, remove acupuncture needle, at operation
Apply and spread penicillin powder, sew up cervical region wound.Blank group only peels off blood vessel, and other is with modeling group.
2.2 Testing index and assay method
2.2.1 Testing index
After all reper-fusion 24h, take a serum and survey the content of NSE, S-100 β.Then take off cervical vertebra to put to death, put rapidly
Taking brain on ice chest, sagittal cuts half and puts in 10% formalin solution, fixes one week, paraffin embedding, makees HE dyeing；Second half is used
Normal saline flushing bloodstain, then exhaust normal saline with filter paper, weigh, the ice saline volume that should add according to this re-computation,
With normal saline (ml): cerebral tissue (g)=9: the ratio of 1, in the large beaker fill ice cube, it is made into the brain of 10% by homogenizer
Homogenate, 3000r/min 4 DEG C, centrifugal 10min, take supernatant in less than-20 DEG C of preservation, press test kit description mensuration brain respectively
Albumen, the content of LD, MDA and LDH, SOD, the vigor of ATP enzyme in homogenate.
2.2.2 the assay method of Coomassie brilliant blue albumen
Coomassie brilliant blue protein determination operation sequence
Shake up, wait 10min, 595nm, 1cm optical path, zeroing, survey absorbance.
Note: A, B, C are respectively standard blank tube, standard pipe, measure pipe
Protein concentration (gprot/l)=(C value A value)/(B value-A value) * 0.563 (standard concentration gprot/l)
2.2.3 the assay method of LD in brain homogenate
LD measures Operations Sequence Chart
Shake up, 530nm, 1cm optical path, zeroing, survey absorbance.
Note: A, B, C are respectively standard blank tube, standard pipe, measure pipe
LD content (mmol/gprot)=(C value-A value) * normal concentration (3mmol/L)/(B value-A value)/sample in tissue
Protein content (gprot/L) in Ben
2.2.4 the assay method of LDH in brain homogenate
The operational approach of cerebral tissue LDH
Shake up, put room temperature and put 3 minutes, 440nm, zeroing, 1cm optical path, survey absorbance.
Note: A: standard pipe B: control tube C: measure pipe D: measure control tube
LDH vigor (U/gprot)=(C value-D value)/(A value-B value) * normal concentration (2mmol/L)/sample egg in tissue
Bai Hanliang (gprot/ml)
2.2.5 the assay method of ATP enzyme
Atpase assay method in brain homogenate
Shake up, centrifugal (4000rpm) 10 minutes, take supernatant 0.2ml and determine phosphorus
Determine phosphorus
Shake up, 37 degree of water-baths 30 minutes, be cooled to room temperature, 660nm, 1cm optical path, distilled water zeroing colorimetric
Note: A, B, C, D pipe is respectively control tube, Na+K+-ATP enzyme pipe, Mg++-ATP enzyme pipe, Ca++-ATP enzyme pipe；
Na in tissue+K+-ATP enzyme, Mg++-ATP enzyme and Ca++-ATP enzyme vigor (μm olPi/mgprot/hour)=(B, C
And D value-A value) diluted sample multiple (2.5) × 6/ sample in/standard value * standard pipe concentration (0.5 μm ol/ml) * reaction system
Protein content (mgprot/ml)
2.2.6 the assay method of total T-SOD in brain homogenate
In brain homogenate, total T-SOD measures sequence list
Shaking up, room temperature 10min, at 550nm, 1cm optical path, distilled water returns to zero, colorimetric.
Note: A: measure pipe B: control tube
Total SOD vigor (U/mgprot)=(B value-A value)/B value/50%* reactant liquor cumulative volume/sampling amount/sample to be tested
Protein concentration (mgprot/ml)
2.2.7 the assay method of MDA in brain homogenate
The operational approach of MDA in brain homogenate
Mixing, 95 DEG C of water-baths 40 minutes, take out flowing water cooling, with the rotating speed of 4000 revs/min, centrifugal 10min, take supernatant
At 532nm, 1cm optical path, distilled water returns to zero, and surveys each pipe OD value.
Note: A: blank tube B: standard pipe C: measure pipe D: control tube
MDA content (nmol/mgprot)=(C value-D value) * standard concentration (10nmol/ml)/(B value-A in tissue
Value)/protein content (mgprot/ml)
2.2.8 the assay method of NSE in serum
2.2.9 the assay method of S-100 β in serum
Cleaning Principle, operating procedure: same to NSE.
3 statistical procedures methods
Data analysis SPSS 17.0 processes, measurement data: average ± standard deviationCompare between each group:
One factor analysis of variance, variance is neat: least significant difference (LSD) method, heterogeneity of variance: Games-Howell method is checked, and grade provides
Material: Ridit checks.
4 experimental results
4.1 the results are shown in Table 1 to the impact of mice repeated cerebral ischemia-reperfusion mortality rate.
The table 1 impact on Cerebral Ischemia-reperfusion in Mice model mortality rate
As seen from the above table, model group mortality rate is the highest, is administered the reduction the most in various degree of each group of mortality rate, and explanation is spent in vain
Herba Hedyotidis Diffusae total flavones can reduce mouse death rate, reduces ischemic region brain tissue impairment, protects cerebral tissue.
4.2 the results are shown in Table 2 to the impact of Cerebral Ischemia-reperfusion in Mice brain homogenate LD, MDA
The table 2 impact on mice repeated cerebral ischemia-reperfusion model tissue LD, MDA content
Note: compare model group, medication group has significant difference (* * P < 0.01), hence it is evident that sex differernce (* P < 0.05)
As seen from the above table, compared with sham operated rats, in model group brain homogenate the content of LD, MDA improve all notable (P <
0.01), modeling success is described；Compared with model group, it is large, medium and small that nimodipine group, brain network lead to group, total flavones of oldenlandia diffusa willd
Dosage group all can significantly reduce the content (P < 0.01) of cerebral tissue LD, MDA, shows that each group of total flavones of oldenlandia diffusa willd is to mouse brain
Reperfusion brain LD, MDA content has significant regulation effect, alleviates the level of lipid peroxidation of cerebral tissue, alleviates breast
The accumulation of acid, especially total flavones of oldenlandia diffusa willd are heavy dose of optimal.
The result that affects of ATP enzyme vigor in repeated cerebral ischemia-reperfusion model mice brain homogenate is shown in 3 by 4.3.
The table 3 impact on Cerebral Ischemia-reperfusion in Mice model cerebral tissue ATP enzyme vigor
Note: compare model group, medication group has significant difference (* * P < 0.01), hence it is evident that sex differernce (* P < 0.05)
As seen from the above table: the Na in the Mice brain tissues of model group+K+-ATP enzyme, Mg++-ATP enzyme, Ca++-ATP enzyme is lived
Force rate sham operated rats significantly reduces (P < 0.01), and the success of cerebral ischemia re-pouring model is described；Compared with model group, nimodipine
Group, brain network lead to group, total flavones of oldenlandia diffusa willd big or middle dosage group significantly raises Na in cerebral tissue+K+-ATP enzyme, Mg++-ATP enzyme,
Ca++-ATP enzyme vigor (P < 0.01)；Na in the significantly raised cerebral tissue of total flavones of oldenlandia diffusa willd small dose group+K+The work of-ATP enzyme
Power (P < 0.05), Mg in notable rising cerebral tissue++-ATP enzyme, Ca++-ATP enzyme vigor (P < 0.01).Experimental result confirms: spend in vain
Herba Hedyotidis Diffusae total flavones is by improving cerebral tissue Na+K+-ATP enzyme, Mg++-ATP enzyme, Ca++The vigor of-ATP enzyme, and then suppression ischemia
The energy metabolism impairment of cerebral tissue, thus improve Cerebral Ischemia-reperfusion in Mice damage.Especially big dose of total flavones of oldenlandia diffusa willd
Amount is optimal.
4.4 are shown in Table 4 to the impact of LDH and SOD vigor in cerebral ischemic model Mice Homogenate repeatedly.
Table 4 is on the impact of LDH and SOD vigor in mice repeatedly cerebral ischemic model brain homogenate
Note: compare model group, medication group has significant difference (* * P < 0.01), hence it is evident that sex differernce (* P < 0.05)
As seen from the above table, compared with sham operated rats, LDH activity and SOD level in the cerebral tissue of model group mice ischemia side
All significantly reduce (P < 0.01), mice repeated cerebral ischemia-reperfusion model modeling success is described；Compare with model group, nimodipine
Group, brain network lead to group, total flavones of oldenlandia diffusa willd big or middle dosage group all can significantly raise activity and the SOD level of LDH in cerebral tissue
(P<0.01)；Total flavones of oldenlandia diffusa willd small dose group can the activity of LDH in significantly raised cerebral tissue；And in Mice brain tissues
The vigor of SOD has the trend (P ＞ 0.05) of substantially rising.Show that total flavones of oldenlandia diffusa willd can significantly raise cerebral ischemia repeatedly
The activity of model mice cerebral tissue LDH, total flavones of oldenlandia diffusa willd big-and-middle dosage group significantly improves the work of cerebral tissue Superoxide dismutase
Property, and then protection cerebral tissue, especially total flavones of oldenlandia diffusa willd heavy dose best results.
4.5 are shown in Table 5 to the impact of NSE, S-100 β content in cerebral ischemic model mice serum repeatedly.
Table 5 is on the impact of NSE, S-100 β content in mice repeatedly cerebral ischemic model serum
Note: compare model group, medication group has significant difference (* * P < 0.01), hence it is evident that sex differernce (* P < 0.05)
Can be seen that from upper table and compare with sham operated rats, in model group mice serum NSE, S-100 β content significantly raise (P <
0.01), repeated cerebral ischemia-reperfusion model modeling success is described；With model group ratio, nimodipine group, brain network lead to group, Agkistrodon
SHECAO total flavones large, medium and small dosage group all can significantly reduce NSE content in serum (P < 0.01)；Nimodipine group, brain network are logical
Group, total flavones of oldenlandia diffusa willd heavy dose group all can significantly reduce the content of S-100 β in serum；In total flavones of oldenlandia diffusa willd
Dosage group substantially reduces the content of S-100 β in serum；Total flavones of oldenlandia diffusa willd small dose group has S-in obvious reduction serum
The trend (P ＞ 0.05) of the content of 100 β.Show total flavones of oldenlandia diffusa willd can alleviate the NSE in ischemic injuries mice serum,
S-100 β content, so protection ischemia cause Mouse Neuron damage and neuronal necrosis, especially total flavones of oldenlandia diffusa willd
Heavy dose of best results.
4.6 are shown in Table 6 to the impact of repeated cerebral ischemia-reperfusion Mice brain tissues hippocampus pathological change.
The table 6 impact on repeated cerebral ischemia-reperfusion Mice brain tissues hippocampus pathological change
" " neurocyte not damaged；"+" the cerebral hippocampus district slight edema of neurocyte, the indivedual degeneration of neuron, structure
Fuzzy, the light dye of endochylema；" ++ " cerebral hippocampus district neurocyte intermediate edema, neuron majority degeneration is the most downright bad, structure mould
Stick with paste, the light dye of endochylema；" +++ " cerebral hippocampus district neurocyte severe edema, neuronal necrosis.
Can be seen by upper table and know: blank group cerebral hippocampus neurocyte not damaged；Model group cerebral neuron is downright bad, Hippocampus god
Through cell severe edema；Nimodipine group cerebral hippocampus minority neuronal degeneration, the slight edema of neurocyte；Brain network logical group brain
The indivedual degeneration of hippocampal neuron, the slight edema of neurocyte；Heavy dose of total flavones of oldenlandia diffusa willd group hippocampal neurons in mouse
Other degeneration, the slight edema of neurocyte；The middle dosage total flavones of oldenlandia diffusa willd indivedual degeneration of group hippocampal neurons in mouse, part god
Through cellular edema, structural fuzzy, the light dye of endochylema；Low dose of total flavones of oldenlandia diffusa willd group hippocampal neurons in mouse large area becomes
Property, the light dye of neurocyte edema, karyopyknosis, endochylema.
Checking through Ridit, model group compares sham operated rats significant statistical significance (P < 0.01), and ischemia-reperfusion is described
Model success.Compared with model group, total flavones of oldenlandia diffusa willd large, medium and small dosage group have obvious statistical significance (P <
0.05)；Result point out: total flavones of oldenlandia diffusa willd can improve ischemia-reperfusion injury model Mice brain tissues hippocampus from damage from
And play the effect of protection cerebral tissue.
4.7 are shown in Table 8 to the impact of repeated cerebral ischemia-reperfusion mouse brain cortical areas pathological change
The table 8 impact on repeated cerebral ischemia-reperfusion Mice brain tissues cortical areas pathological change
" " cerebral cortex district not damaged；"+" cerebral cortex district neuron downright bad, neurocyte edema individually；" ++ " is big
The neuron major part necrosis of Cerebral cortex district and neurocyte edema；" +++ " cerebral cortex district severe edema, neuronal necrosis.
Blank group Neurons of Cerebral Cortex not damaged is can be seen that from upper table；Model group cerebral cortex neurons major part
Downright bad, neurocyte severe edema；Nimodipine group cerebral cortex neurons small part degeneration, the slight edema of neurocyte, knot
Structure obscures, the light dye of endochylema；Brain network logical group cerebral cortex neurons small part degeneration, the slight edema of neurocyte, structural fuzzy, born of the same parents
Starch light dye；Heavy dose of total flavones of oldenlandia diffusa willd group cerebral cortex minority neuronal degeneration, neurocyte edema, structural fuzzy,
The light dye of endochylema；Middle dosage total flavones of oldenlandia diffusa willd group cerebral cortex extremely indivedual neuronal degeneration, neurocyte edema, structure mould
Paste, the light dye of endochylema；Low dose of total flavones of oldenlandia diffusa willd cerebral cortex neurons major part degeneration is even downright bad, neurocyte is tight
Heavy water is swollen, the light dye of endochylema, structural fuzzy.
Analyzing through Ridit, model group compares sham operated rats significant statistical significance (P < 0.01), illustrates that modeling becomes
Merit；Brain network leads to group, nimodipine group, large, medium and small dosage total flavones of oldenlandia diffusa willd group have significantly statistics compared with model group
Learn meaning (P < 0.01), illustrate each medication group all can significantly protect ischemia-reperfusion Mice brain tissues cortical areas from damage, because of
This, do pathological observation and have diagnostic significance intuitively experiment mice.
5 conclusions
Changed by the pathology damage observing modeling Mice brain tissues hippocampus and cortical areas, mortality rate, NSE in serum,
The changes of contents of S-100 β, the ATP enzyme vigor in brain homogenate, the activity of SOD, LDH, prompting total flavones of oldenlandia diffusa willd can reduce
The mortality rate of animal；Reduce the content of LD, MDA, NSE, S-100 β；Improve ATP enzyme, the vigor of SOD, LDH；Alleviate Ischemia Reperfusion
The free radical caused after note generates and increases；Alleviate because of the too high damage to cerebral tissue of acidity；Improve energy metabolism impairment, reduce by
The brain tissue impairment cascade reaction that this causes, improves the neural cell injury of cerebral tissue cortical areas and hippocampus, with Herba Hedyotidis Diffusae
Grass total flavones heavy dose group is optimal.
Claims (1)
1. a total flavones of oldenlandia diffusa willd is that sole active agent is in preparation treatment meninges microcirculation disturbance, brain ischemia medicament
Application, described total flavones of oldenlandia diffusa willd is, Herba Hedyotidis Diffusae was ground into the coarse powder of 30 mesh sieves, first adds coarse powder 10 times
Weight petroleum ether reflux, extract, 1h, defat, discard petroleum ether, obtain medicinal residues；Medicinal residues volatilize petroleum ether, add 10 times of weight, matter every time
Amount concentration 80% soak with ethanol 0.5h, reflux, extract, 2 times, each 1h, filter, merging filtrate, decompression recycling ethanol to without alcohol taste,
Add distilled water to be dispersed into and be equivalent to, containing crude drug amount 0.3g/ml, obtain sample liquid；AB-8 type macroporous adsorbent resin in sample liquid, first with 2
Times column volume distilled water eluting, discards water liquid；Continue with 4 times of column volumes, mass concentration 10% ethanol elution, discard eluent；Use again
6 times of column volumes, mass concentration 80% ethanol elutions, collect 80% ethanol elution, and decompression recycling ethanol is to without alcohol taste, freezing dry
Dry, pulverize, obtain total flavones of oldenlandia diffusa willd.
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Application publication date: 20161012 |