CN102068483B - Persimmon leaf extract for improving brain ischemic tolerance - Google Patents
Persimmon leaf extract for improving brain ischemic tolerance Download PDFInfo
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- CN102068483B CN102068483B CN2011100031133A CN201110003113A CN102068483B CN 102068483 B CN102068483 B CN 102068483B CN 2011100031133 A CN2011100031133 A CN 2011100031133A CN 201110003113 A CN201110003113 A CN 201110003113A CN 102068483 B CN102068483 B CN 102068483B
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Abstract
The invention relates to a persimmon leaf extract for improving brain ischemic tolerance, which can effectively solve the problem of improvement in the brain ischemic tolerance. The technical scheme adopted by the invention is that: the persimmon leaf extract is prepared by the following steps of: crushing persimmon leaves into coarse powder, adding ethanol at the mass concentration of 70 percent and in an amount which is 8 to 10 times the weight of the persimmon leaf coarse powder, performing impregnation for 30min, and performing refluxing and extraction for 1h; and performing filtration, adding the ethanol at the mass concentration of 70 percent and in an amount which is 6 to 8 times the weight of the persimmon leaf coarse powder, performing refluxing and extraction for 1h, performing filtration, mixing the filtrate, recycling the ethanol, adjusting the pH to 13 by using NaOH, and performing centrifugation at 3,000 r/min for 15min to obtain the persimmon leaf extract, or weighing the persimmon leaf coarse powder, and performing extraction for 3h by using a supercritical CO2 extraction method under the conditions of extraction pressure of 30.0mPa, extraction temperature of 50 DEG C and CO2 flow rate of 30Kg/h to obtain the persimmon leaf extract. The extract provided by the invention is effectively used for preparing medicaments for improving the brain ischemic tolerance, is prepared from rich raw materials by a simple method, has low cost and good effects, develops the medicinal value of the persimmon leaves and is an innovation in Chinese medicament.
Description
One, technical field
The present invention relates to medicine, improve the extract of cerebral ischemia tolerance effect in particularly a kind of Folium Kaki.
Two, background technology
Diospyros Ebenaceae (Ebenaceae) Diospyros (Diospyros) plant Fructus Kaki (Diospyros kaki L..f) plant is the edible variety of various places, China north and south common cultivated.Fructus Kaki, Fructus Kaki nuclear, Calyx Kaki, Mannosum Kaki, Fructus kaki Immaturus preparatum, Fructus kaki Siccus, persimmon heart wood, Flos Kaki, Folium Kaki, Pericarpium Kaki, Cortex Kaki and Radix Kaki skin etc. all can be used as and medicinally maybe might become medicine.The Folium Kaki resource is abundant especially in China, and is easy to use, and medical determined curative effect receives people's attention just day by day, the fresh or dried leaves of Fructus Kaki, and its beginning of being used as medicine is shown in bright " the southern regions of the Yunnan Province book on Chinese herbal medicine " record and " applies the swelling skin ulcer through red leaves ".Contain rich nutrient substances and functional component in the Folium Kaki; Have plurality of health care functions such as anti-inflammation, promoting the production of body fluid to quench thirst, heat-clearing and toxic substances removing, lung moistening heart tonifying, antitussive hemostasis, anti-cancer and cancer-preventing; And Folium Kaki product edible safety is nontoxic, so the exploitation prospect of Folium Kaki is wide.
Modern study shows: it is the aromatic (naphthoquinone class, aphthols) of skeleton that Folium Kaki contains with the naphthalene; Benzofurantone (flavonoid, anthocyan, Coumarins); Terpenoid (mainly be triterpenoid and all be the pentacyclic triterpene chemical compound), sitosterol class, fatty acid; Tannin contains a large amount of vitamin Cs, E and organic acid etc. in the Folium Kaki.Folium Kaki has the good cardio and vascular function effect that improves.Folium Kaki has anastalsis; The Folium Kaki extracting solution has antibiotic, refrigeration function; Folium Kaki has tangible antioxidation; Folium Kaki has tangible fat-reducing and antihyperglycemic; Folium Kaki has anticancer change and antimutagenesis, regulates immunity function.The Folium Kaki powder can significantly reduce the cholesterol concentration in the edible high sterin food mouse liver, increases the drainage of bile acid.
Folium Kaki has vessel softening and the hardened effect of prevention of arterial clinically, is used to treat cardiovascular and cerebrovascular disease; Use Folium Kaki and can treat multiple hemorrhage; Folium Kaki has anti-inflammation, refrigeration function; Folium Kaki has the effect of prevention anemia, lowering blood-fat and reducing weight and beauty treatment; Have anticancer and antimutagenesis; The Folium Kaki compound recipe is effective to some disease of women such as dysmenorrhea, and persimmon leaf tea has patient's the mental status and appetite.Folium Kaki has the effect of the diuresis of purifying the blood, treatment burn and decubital ulcer.
(Brain ischemic tolerance BIT) is meant that in advance transience ischemia or slight hypoxia excite or mobilize the intrinsic protective capacities of body, make body produce the phenomenon of defence and protective effect to serious subsequently ischemia, anoxia to cerebral ischemia tolerance phenomenon [1].Cerebral ischemia tolerance phenomenon is found when the research myocardial ischemia at first, has also been found similar phenomenon in the organs such as brain, skeletal muscle, liver, kidney, small intestinal.Think that at present the mechanism of cerebral ischemia tolerance mainly contains excitatory amino acid (EAA), inflammation, heat shock protein (HSP), adenosine, apoptosis, reactive astrocyte hypertrophy and signal transduction pathway.Medicine (adenosine, 3-nitropropionic acid (3-NPA), some antibiotic, anaesthetic, aspirin, endotoxin, Kallidin I, neurotrophic factor etc.) is to the raising effect of cerebral ischemia tolerance; But because of reason in many ways, said medicine is used in the cerebral ischemia tolerance and has been received restriction.Motherland's medical science does not have the clear and definite lifting manipulation of cerebral ischemia tolerance, but motherland's medical science is dialectical clear to cerebral ischemia, and cerebral ischemia belongs to the category of the traditional Chinese medical science " ischemia apoplexy ".The generation of ischemia apoplexy is main relevant with factors such as wind, fire, expectorant, silt, void, and the closest with " silt " relation, " activating blood circulation to dissipate blood stasis " improves the basic skills of cerebral ischemia tolerance.So how utilize the big treasure-house of Chinese medicine to solve and improve cerebral ischemia tolerance effect? So far do not see open report is arranged.
Three, summary of the invention
To above-mentioned situation, for overcoming the defective of prior art, the present invention's purpose just provides the extract that improves cerebral ischemia tolerance effect in a kind of Folium Kaki, can effectively solve the problem that improves cerebral ischemia tolerance effect.
The technical scheme that the present invention solves is, this extract is a Folium Kaki extract, is that Folium Kaki is ground into coarse powder, adds mass concentration that the 8-10 of Folium Kaki coarse powder weight doubly measures and be 70% ethanol, soak 30min after, reflux, extract, 1h; Filter, the 6-8 mass concentration doubly that medicinal residues add Folium Kaki coarse powder weight again is 70% ethanol, and reflux, extract, 1h filters, and merges filtrating twice; After reclaiming ethanol, transfer pH to 13, the centrifugal 15min of 3000r/min with NaOH; Get Folium Kaki extract, or take by weighing the Folium Kaki coarse powder, use supercritical CO
2Extraction, extracting pressure are 30.0mPa, 50 ℃ of extraction temperature, CO
2Flow 30Kg/h, extraction 3h gets Folium Kaki extract.
Extract of the present invention is effective to prepare the medicine that treatment improves cerebral ischemia tolerance effect.Method for preparing is simple, abundant raw materials, and cost is low, and is effective, opened up the medical value of Folium Kaki, is the innovation on the Chinese medicine.
Four, the specific embodiment
Below in conjunction with embodiment, specific embodiments of the invention elaborates.
Embodiment 1, and the present invention is to pulverize 30 mesh sieves by Folium Kaki through pulverizer to become the Folium Kaki coarse powder in the specific implementation; Then, the mass concentration of getting ten times of amounts of Folium Kaki coarse powder 100 gram adding Folium Kaki coarse powder weight is 70% ethanol, 1000 grams, soaks 30 minutes; Reflux, extract, 1 hour is crossed and is filtered filtrating; The mass concentration that medicinal residues add 8 times of Folium Kaki coarse powder weight again is 70% ethanol 800g, and reflux, extract, 1 hour is crossed and filtered filtrating, merges filtrating twice, reclaim ethanol after, using the NaOH adjust pH is 13, the centrifugal 15min of 3000r/min, Folium Kaki extract.
Embodiment 2; The present invention also can be by following realization: Folium Kaki was pulverized 50 mesh sieves through pulverizer, become the Folium Kaki coarse powder then in practical implementation; The mass concentration of getting 8 times of amounts of Folium Kaki coarse powder 50 gram adding Folium Kaki coarse powder weight is 70% ethanol, 400 grams; Soaked 30 minutes, reflux, extract, 1 hour is crossed and is filtered filtrating; The mass concentration that medicinal residues add 6 times of Folium Kaki coarse powder weight again is 70% ethanol 300g, and reflux, extract, 1 hour is crossed and filtered filtrating, merges filtrating twice, reclaim ethanol after, using the NaOH adjust pH is 13, the centrifugal 15min of 3000r/min, Folium Kaki extract.
Embodiment 3, and the present invention also can be by following realization: Folium Kaki is ground into coarse powder through pulverizer in practical implementation; Then, the mass concentration of getting 9 times of amounts of Folium Kaki coarse powder 80 gram adding Folium Kaki coarse powder weight is 70% ethanol, 720 grams, soaks 30 minutes; Reflux, extract, 1 hour is crossed and is filtered filtrating; The mass concentration that medicinal residues add 7 times of Folium Kaki coarse powder weight again is 70% ethanol 560g, and reflux, extract, 1 hour is crossed and filtered filtrating, merges filtrating twice, reclaim ethanol after, using the NaOH adjust pH is 13, the centrifugal 15min of 3000r/min, Folium Kaki extract.
Embodiment 4, and the present invention also can be realized by following concrete scheme: Folium Kaki is ground into coarse powder through pulverizer, gets coarse powder 100g, insert in the supercritical extraction reactor, use supercritical CO in practical implementation
2Extraction, extracting pressure are 30.0mPa, 50 ℃ of temperature, CO
2Flow 30Kg/h, extraction 3h gets Folium Kaki extract.
Extract of the present invention can be effective to prepare the medicine that treatment improves cerebral ischemia tolerance effect, has obtained sufficient proof for improving cerebral ischemia tolerance effect through test, and relevant testing data is following:
Influence to mouse brain ischemia tolerance model
1 experiment material
1.1 reagent chemicals
Folium Kaki extract of the present invention: take by weighing the Folium Kaki coarse powder, add 70% ethanol that 8-10 doubly measures, reflux, extract, 1h behind the immersion 30min; Filter, medicinal residues add 70% ethanol that 6-8 doubly measures again, and reflux, extract, 1h filters.Merge filtrating twice, transfer pH to 13 with NaOH behind the recovery ethanol, centrifugal (3000r/min) 15min gets Folium Kaki extract.Or take by weighing the Folium Kaki coarse powder, and adopting the supercritical CO 2 extraction, extracting pressure is 30.0mPa, 50 ℃ of extraction temperature, CO
2Flow 30Kgh
-1, extraction time 3h gets Folium Kaki extract.
Folium Ginkgo extract sheet (Ginaton), German Weil-McLain doctor Shu pharmaceutical factory, lot number 9900908; Chloral hydrate, Tianjin section close europeanized reagent development centre, lot number 20071018; Medical alcohol, the bright great industry and trade company limited in Zhengzhou, lot number 20090801; Sodium chloride injection, Zhengzhou Yonghe Pharmaceutical Co, lot number 09073105; Iodine [
125I] 6-ketone-prostaglandin-F
1The α Endothelin, Beijing pul great achievement bio tech ltd PLA General Hospital Science and Technology Development Center puts and exempts from institute, lot number 20091125; Iodine [
125I] the plain B of thromboxane
2The radioimmunoassay, RIA medicine box, Beijing pul great achievement bio tech ltd PLA General Hospital Science and Technology Development Center puts and exempts from institute, lot number 20091125; Plasminogen activator (t-PA) assay test kit (ELISA), Shanghai Sun Bio-Tech Co., Ltd., lot number 41130; Plasminogen activator inhibitor-1 (PAI-1) assay test kit (ELISA), Shanghai bio tech ltd, lot number 42135.
1.2 experimental apparatus
The JT6001 electronic balance, the sub-instrument plant of last Nereid's atmospheric electricity; 1202080019 electronic analytical balances, Ao Haosi (Shanghai) company; The KDC-160HR High speed refrigerated centrifuge, good branch company in the Keda Innovation Co., Ltd; Enzyme is exempted from appearance, 680MicroplateReader, Bio-Rad Laboratories; SN-695B type intelligence is put and is exempted from the γ measuring instrument, day ring instrument one factory of Shanghai nuclear research institute.
1.3 laboratory animal
Cleaning level mice, the Kunming kind, male, body weight 25～30g; Quality certification numbering 911137 is provided by Hebei province's Experimental Animal Center.
2. experimental technique
2.1 modeling and administration
Get 126 of mices, evenly be divided into 7 groups at random, be respectively dose groups, Folium Kaki ethanol extract low dose group, Folium Ginkgo extract group in sham operated rats, ischemia-reperfusion group, pretreated model group, Folium Kaki ethanol extract high dose group, the Folium Kaki ethanol extract.All mices face upward and are fixed on the operating-table through 10% chloral hydrate (0.03ml/10g) intraperitoneal injection of anesthesia, the throat median incision, and passivity is separated bilateral common carotid arteries.Except that sham operated rats, ischemia-reperfusion group, all the other are respectively organized mice and close bilateral common carotid arteries with arteriole folder folder, and blocking blood flow 10min recovers perfusion then respectively; Sham operated rats, ischemia-reperfusion group only expose bilateral common carotid arteries, but blocking blood flow not.After animal surgery is revived; Each administration group gives high dose Folium Kaki ethanol extract (400mg/kg), middle dosage Folium Kaki ethanol extract (200mg/kg), low dosage Folium Kaki ethanol extract (100mg/kg), Folium Ginkgo extract sheet (40mg/kg) respectively; Sham operated rats, ischemia-reperfusion group and pretreated model group are irritated the normal saline with volume; The perfusion volume of all mices is 0.1ml/10g, administration every day 1 time, successive administration 5 days.1h after fasting 12h administration in the 5th day, all mices face upward the position and are fixed on the operating-table through 10% chloral hydrate (0.03ml/10g) intraperitoneal injection of anesthesia; The throat median incision; Passivity is separated bilateral common carotid arteries, and except that sham operated rats, all the other mices all close bilateral common carotid arteries with arteriole folder folder; Blocking blood flow 30min recovers perfusion then respectively; Sham operated rats only exposes bilateral common carotid arteries, not blocking blood flow.Keep 24 ± 1 ℃ of ambient temperatures in the art.Dead 33 mices in the experimentation, each is organized the mice situation and sees the result.
2.2 observation item and detection method
All mices are got blood in last operation back 24h, the sodium citrate anticoagulant, and separated plasma adopts ELISA to measure plasminogen activator (t-PA) content, plasminogen activator inhibitor-1 (PAI-1) content; Put to death after getting blood, strip cerebral tissue rapidly, crown cutting done HE dyeing, cuts the 15mg of cortical tissue, adopts measured by radioimmunoassay 6-ketone-prostaglandin-F
1α, the plain B of thromboxane
2Content.
2.2.1 plasminogen activator Determination on content method
Inspection principle: adopt enzyme linked immunological absorption double antibody sandwich method principle quantitative assay tissue plasminogen activator level.Encapsulate people t-PA antibody and combine with t-PA in the sample to be tested, add enzyme labelled antibody and form complex, the latter and substrate-function present chromogenic reaction.The A value that the 490nm place records is directly proportional with sample to be tested t-PA content.
Operation sequence: 1. reagent is rebuild dense diluent with being prepended to 37 ℃ of water-bath 15min, and jolting is (because the mother solution salinity is high, refrigerator is preserved to be prone to freeze and separated out) evenly, does 10 times of dilutions (the dense diluent of 1ml+9ml distilled water) with distilled water then.Dense cleaning mixture is with being prepended to 37 ℃ of water-bath 15min, and jolting is (because the mother solution salinity is high, refrigerator is preserved to be prone to freeze and separated out) evenly, does 20 times of dilutions (the dense diluent of 1ml+19ml distilled water) with distilled water then.Every bottle of calibration object is accurately redissolved (60ng/ml) with the 1.8ml diluent.Get 250 μ l, make four doubling dilutions with diluent, concentration be 60,30,15,7.5, five standard points of 3.75ngml.2. the every hole of application of sample adds variable concentrations calibration object or sample to be tested 100 μ l, and the blank hole adds diluent 100 μ l, hatches 150 minutes for 37 ℃.3. washing discards liquid in the reacting hole, fills with each hole with cleaning mixture, leaves standstill for 3 seconds, dries, and claps after three times repeatedly and does.4. add the every hole of enzyme labelled antibody and add enzyme labelled antibody 100 μ l, hatched 60 minutes for 37 ℃.5. washing discards liquid in the reacting hole, fills with each hole with cleaning mixture, leaves standstill for 3 seconds, dries, and claps after three times repeatedly and does.6. every OPD dissolved with the 5ml substrate buffer solution before usefulness was faced in colour developing.Every hole adds substrate solution 100 μ l, hatches 15 minutes for 37 ℃.7. stop every hole and add stop buffer 50 μ l.8. colorimetric 490nm place on ELIASA with blank air-conditioning zero, measures each hole A value.9. date processing is with A
490T-PA calibration object concentration (ng/ml) is made standard curve on common coordinate, sample to be tested t-PA content (ng/ml) can be found from standard curve.
2.2.2 plasminogen activator inhibitor-1 Determination on content method
Inspection principle: adopt enzyme linked immunological absorption double antibody sandwich method principle quantitative assay plasminogen activator inhibitor-1 level.Encapsulate people PAI-1 antibody and combine with PAI-1 in the sample to be tested, add enzyme labelled antibody and form complex, the latter and substrate-function present chromogenic reaction.The A value that the 490nm place records is directly proportional with sample to be tested PAI-1 content.
Operation sequence: 1. reagent is rebuild dense diluent with being prepended to 37 ℃ of water-bath 15min, and jolting is (because the mother solution salinity is high, refrigerator is preserved to be prone to freeze and separated out) evenly, does 10 times of dilutions (the dense diluent of 1ml+9ml distilled water) with distilled water then.Dense cleaning mixture is with being prepended to 37 ℃ of water-bath 15min, and jolting is (because the mother solution salinity is high, refrigerator is preserved to be prone to freeze and separated out) evenly, does 20 times of dilutions (the dense diluent of 1ml+19ml distilled water) with distilled water then.Every bottle of calibration object is accurately redissolved (120ng/ml) with the 0.5ml diluent.Get 250 μ l, make six doubling dilutions with diluent, concentration be 120,60,30,15,7.5,3.75, seven standard points of 1.875ngml.2. the every hole of application of sample adds variable concentrations calibration object or sample to be tested 100 μ l, and the blank hole adds diluent 100 μ l, hatches 150 minutes for 37 ℃.3. washing discards liquid in the reacting hole, fills with each hole with cleaning mixture, leaves standstill for 3 seconds, dries, and claps after three times repeatedly and does.4. add the every hole of enzyme labelled antibody and add enzyme labelled antibody 100 μ l, hatched 60 minutes for 37 ℃.5. washing discards liquid in the reacting hole, fills with each hole with cleaning mixture, leaves standstill for 3 seconds, dries, and claps after three times repeatedly and does.6. every OPD dissolved with the 5ml substrate buffer solution before usefulness was faced in colour developing.Every hole adds substrate solution 100 μ l, hatches 15 for 37 ℃.7. stop every hole and add stop buffer 50 μ l.8. colorimetric 490nm place on ELIASA with blank air-conditioning zero, measures each hole A value.9. date processing is with A
490PAI-1 calibration object concentration (ng/ml) is made standard curve on common coordinate, sample to be tested PAI-1 content (ng/ml) can be found from standard curve.
2.2.3 thromboxane B
2Assay method
Measuring principle: utilize the liquid phase competition to suppress principle, adopt counterbalanced procedure that sample is measured.Sample to be tested or standard substance add being at war with property association reaction together with antiserum of limiting the quantity of and labelled antigen; After reacting completely, add immune release agent, isolate antigen-antibody complex; Measure the radioactivity (B) of complex, calculate the combination rate (B/B0%) of each standard pipe.
Calibration reagent: buffer A, 1 bottle (liquid); Buffer B, 1 bottle (blueness, liquid); Antiserum: 1 bottle (white, lyophilized powder); Standard substance: 1 (white, lyophilized powder), concentration are that concentration is 20,60,180,500,1000pg/ml;
125I-TXB
2: 1 bottle (white, lyophilized powder); The PR separating medium: 1 bottle (liquid) fully shakes up before the use.
Operation sequence: counterbalanced procedure is adopted in this experiment, gets polyethylene test tube numbering, the according to the form below procedure operation:
TXB
2RIA liquid feeding program (μ l)
Date processing: the program of working out in advance with the γ calculating instrument has directly provided related parameter, calibration curve and sample concentration.
2.2.46-ketone-PGF
1The assay method of α
Measuring principle: utilize the liquid phase competition to suppress principle, adopt counterbalanced procedure that sample is measured.Sample to be tested or standard substance and the antiserum of limiting the quantity of add react a period of time together after; Add again and being at war with property of labelled antigen association reaction; After reacting completely, add immune release agent, isolate antigen-antibody complex; Measure the radioactivity (B) of complex, calculate the combination rate (B/B0%) of each standard pipe.
Calibration reagent: buffer A, 1 bottle (liquid); Buffer B, 1 bottle (redness, liquid); Antiserum: 1 bottle (white, lyophilized powder); Standard substance: 1 (white, lyophilized powder), concentration are that concentration is 20,60,180,500,1000pg/ml;
125I-6-Keto-PGF
1α: 1 bottle (white, lyophilized powder); The PR separating medium: 1 bottle (liquid) fully shakes up before the use.
Operation sequence: non-weighing apparatus method is adopted in this experiment, gets polyethylene test tube numbering, the according to the form below procedure operation:
6-Keto-PGF
1α RIA liquid feeding program (μ l)
Date processing: the program of working out in advance with the γ calculating instrument has directly provided related parameter, calibration curve and sample concentration.
3. experimental result
3.1 influence to cerebral ischemia tolerance mice plasma t-PA, PAI-1 level
The result sees table 1.
Table 1 Folium Kaki ethanol extract is to the influence
of cerebral ischemia tolerance mice plasma t-PA, PAI-1 level
Annotate: compare with sham operated rats
△ △P＜0.01,
△P＜0.05; Compare with ischemia-reperfusion group
▲ ▲P＜0.01,
▲P＜0.05; Compare with model group
*P＜0.01,
*P＜0.05
Last table can be found out, compares with sham operated rats, and ischemia-reperfusion group and pretreated model group mice plasma t-PA concentration, PAI-1 concentration have significant statistical significance (P＜0.01); Compare with ischemia-reperfusion group, pretreated model group mice plasma t-PA concentration, PAI-A concentration have notable difference (P＜0.05), explain that the ischemia pretreatment can produce toleration to the serious cerebral ischemia that takes place once more, the modeling success.Compare with the pretreated model group; Folium Kaki ethanol extract low dose group mice plasma t-PA concentration, PAI-1 concentration do not have statistical significance (P＞0.05); The middle and high dose groups of Folium Kaki ethanol extract; Positive controls mice plasma t-PA concentration difference is (P＜0.01) significantly, dose groups mice plasma PAI-1 concentration difference obvious (P＜0.05) in the Folium Kaki ethanol extract, and Folium Kaki ethanol extract high dose group, Folium Ginkgo extract group mice plasma PAI-1 concentration difference be (P＜0.01) significantly.
Results suggest: according to above-mentioned Mathematical Statistics Analysis; This laboratory animal pretreated model group is compared with ischemia-reperfusion group, and the damage of ischemia-reperfusion group will weigh, and each group of medication all has the effect that strengthens the anti-ischemia of brain cell in various degree; Folium Kaki ethanol extract high dose group, the effect of Folium Ginkgo extract group are best; Secondly be dose groups in the Folium Kaki ethanol extract, though Folium Kaki ethanol extract low dose group not statistically significant has improved cerebral ischemia tolerance effect to a certain extent.
3.2 to cerebral ischemia tolerance mouse brain cortex homogenate TXB
2, 6-Keto-PGF
1α and TXB
2/ 6-Keto-PGF
1The result that influences of alpha levels sees table 2.
Table 2 Folium Kaki ethanol extract tolerates mouse brain to cerebral ischemia
Cortex homogenate TXB
2, 6-Keto-PGF
1The influence of alpha levels
Annotate: compare with sham operated rats
△ △P＜0.01,
△P＜0.05; Compare with the ischemic injuries group
▲ ▲P＜0.01,
▲P＜0.01; Compare with model group
*P＜0.01,
*P＜0.05
Table 3 Folium Kaki ethanol extract is to cerebral ischemia tolerance mouse brain cortex
TXB
2/ 6-Keto-PGF
1The influence of alpha levels
Annotate: compare with sham operated rats
△ △P＜0.01,
△P＜0.05; Compare with the ischemic injuries group
▲ ▲P＜0.01,
▲P＜0.01; Compare with model group
*P＜0.01,
*P＜0.05
Can intuitively find out by last table, compare, ischemia-reperfusion group and pretreated model group mouse brain cortex 6-Keto-PGF with sham operated rats
1α, TXB
2Content and TXB
2/ 6-Keto-PGF
1α has significant statistical significance (P＜0.01); Compare model control group mice blood cerebral cortex 6-ketone-prostaglandin-F with ischemia-reperfusion group
1α, the plain B of thromboxane
2Content and TXB
2/ 6-Keto-PGF
1α significant difference (P＜0.01) explains that the ischemia pretreatment can produce toleration to the serious cerebral ischemia that takes place once more, the modeling success.Compare the basic, normal, high dose groups of Folium Kaki ethanol extract, positive controls mouse brain cortex 6-ketone-prostaglandin-F with model control group
1α, the plain B of thromboxane
2Content and TXB
2/ 6-Keto-PGF
1α significant difference (P＜0.01).
Results suggest: according to above-mentioned Mathematical Statistics Analysis; This laboratory animal cerebral ischemia tolerance pretreated model group is compared with ischemia-reperfusion group; The damage of ischemia-reperfusion group will weigh, and each group of medication all has can alleviate the damage that cerebral ischemia re-pouring causes, and has improved the generation of the anti-ischemia of brain.
3.3 the coordinating protection exercising result to mouse brain ischemia tolerance model cerebral tissue is seen table 4.
Table 4 Folium Kaki ethanol extract is to the coordinating protection effect of mouse brain ischemia tolerance model cerebral tissue
Different pathological changes occurring according to the laboratory animal cranial nerve cell can be with being divided into level Four: "-" cranial nerve cell body is big, endochylema abundant, nucleus is all normal; "+" few part cranial nerve cell volume-diminished, endochylema reduces, and nucleus is normal basically; The atrophy of " ++ " part cranial nerve cell, endochylema reduce, nucleus is light dyes or disappears; " +++" the obvious atrophy of most of laboratory animal cranial nerve cell, endochylema obviously reduces, nucleus is smudgy or disappearance.
Sham operated rats animal brain neurocyte, cell membrane, kernel are high-visible; The obvious swelling of ischemia-reperfusion group animal brain neurocyte visible cell, cell volume increase, or pyknosis occurs, and endochylema, nucleus indigo plant are dyed phenomenon; Pretreated model group cranial nerve cell visible part cellular swelling, the light phenomenon of dying of nuclear; Folium Kaki ethanol extract low dose group animal brain neurocyte visible part cellular swelling, the light phenomenon of dying of nuclear; Visible few part cellular swelling of dose groups animal brain neurocyte, the light phenomenon of dying of nuclear in the Folium Kaki ethanol extract; The visible individual cells swelling of Folium Kaki ethanol extract high dose group animal brain neurocyte, the light phenomenon of dying of nuclear; The visible individual cells swelling of Folium Ginkgo extract treated animal neurocyte, the light phenomenon of dying of nuclear.
Through rank test, with the sham operated rats ratio, ischemia-reperfusion group and pretreated model group have significant statistical significance (P＜0.01); Compare with ischemia-reperfusion group, the pretreated model group has notable difference (P＜0.05).Compare with the pretreated model group, low, the middle dose groups of Folium Kaki ethanol extract has notable difference (P＜0.05), and Folium Kaki ethanol extract high dose group, there were significant differences for the Folium Ginkgo extract group (P＜0.01).
Results suggest: according to pathological observation result and above-mentioned Mathematical Statistics Analysis; This laboratory animal cerebral ischemia tolerance pretreated model group is compared with ischemia-reperfusion group; The pathological change of ischemia-reperfusion group will weigh; The obvious atrophy of cranial nerve cell, endochylema obviously reduces, the nucleus mould is light dyes, sticks with paste unclear or disappearance; Each group of medication all has the different effects that strengthen the anti-ischemia of brain cells, and Folium Kaki ethanol extract high dose group and the effect of Folium Ginkgo extract group are best, secondly is in the Folium Kaki ethanol extract, low dose group.
4 conclusions
Above-mentioned test through repeatedly (carrying out 12 times) in 2 years; All obtained identical or akin result; This fully shows the Folium Kaki ethanol extract t-PA content in the cerebral ischemic model mice plasma that can obviously raise; Reduce PAI-1 content, explain that the Folium Kaki ethanol extract can well improve cerebral tissue fibrinolytic system function; Rising mouse brain cortex 6-Keto-PGF
1Alpha content, reduction TXB
2Content.The Folium Kaki ethanol extract has good cerebral ischemia tolerance synergism.
Claims (1)
1. a Folium Kaki extract improves the application in the cerebral ischemia tolerance drugs with function in the preparation treatment; Described this extract is that Folium Kaki is ground into coarse powder; The mass concentration that the 8-10 of adding Folium Kaki coarse powder weight doubly measures is 70% ethanol, behind the immersion 30min, and reflux, extract, 1h; Filter, the 6-8 mass concentration doubly that medicinal residues add Folium Kaki coarse powder weight again is 70% ethanol, and reflux, extract, 1h filters, and merges filtrating twice, reclaim ethanol after, transfer pH to 13 with NaOH, the centrifugal 15min of 3000r/min, Folium Kaki extract.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101637492A (en) * | 2009-08-31 | 2010-02-03 | 陕西师范大学 | Preparation method of total flavonoids in persimmon leaves and application of total flavonoids in liver-protecting drugs and health foods |
| CN101744863A (en) * | 2010-01-29 | 2010-06-23 | 广州白云山和记黄埔中药有限公司 | Persimmon leaf extract preparation, preparing method thereof and application thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101637492A (en) * | 2009-08-31 | 2010-02-03 | 陕西师范大学 | Preparation method of total flavonoids in persimmon leaves and application of total flavonoids in liver-protecting drugs and health foods |
| CN101744863A (en) * | 2010-01-29 | 2010-06-23 | 广州白云山和记黄埔中药有限公司 | Persimmon leaf extract preparation, preparing method thereof and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 岳红等.超临界二氧化碳萃取柿叶黄酮的工艺研究.《化学研究与应用》.2005,(第03期), * |
| 晏亦林等.柿叶总黄酮提取方法研究.《中药材》.2003,(第11期), * |
| 贝伟剑等.柿叶黄酮对大鼠急性前脑缺血再灌注损伤的保护作用.《广西中医药》.2009,(第02期), * |
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