CN101204438B - Application of potentilla anseriana extractive on preparating anti-ischemic - Google Patents
Application of potentilla anseriana extractive on preparating anti-ischemic Download PDFInfo
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- CN101204438B CN101204438B CN2006101304387A CN200610130438A CN101204438B CN 101204438 B CN101204438 B CN 101204438B CN 2006101304387 A CN2006101304387 A CN 2006101304387A CN 200610130438 A CN200610130438 A CN 200610130438A CN 101204438 B CN101204438 B CN 101204438B
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Abstract
The invention relates to an extractive of a potentilla anserina in the application of preparing the medicine for curing ischemic disease of heart and brain, and curing ischemia reperfusion injury of heart and brain. The ischemia and the ischemia reperfusion injury are the diseases which are seriously affected the health of human beings. With the change in the ageing of population and the life style of people, the category of disease has become a major disease which threatens the life of human being, especially the ischemia reperfusion injury is the common complicating disease of the percutaneous intervention of the present clinical cardiovascular PCI and bypass grafting. In the recent years, the scholars both abroad and at home take a lot of researches on using traditional Chinese medicine to cure the diseases of the ischemia and the ischemia reperfusion injury, and the researches show that the traditional Chinese medicine has strong tolerance to oxygen-poor organism and has good protection for the ischemia and the ischemia reperfusion injury. The extractive of the potentilla anserina in the invention proves that the traditional Chinese medicine has remarkable effects in curing the diseases of the ischemia and the ischemia reperfusion injury.
Description
Technical field
The present invention relates to a kind of pharmaceutical preparation, these goods are made up of Radix potentillae anserinae extract and medicine acceptable carrier, and these goods can be used for preventing and treating ischemic diseases.
Background technology
Radix potentillae anserinae (Potentilla anserina L.) is the underground tuber of the little section of rose potentilla plants potentilla anserina, have the laudatory title of " Herba Herminii ", and the Tibetan medicine is called " Zhuo Laosa once ".It mainly grows in grassy marshland, the rivers and canals next door of Qinghai-Tibet height above sea level 2600m-4750m, is that a kind of distribution is extensive, big, the natural fine wild plant resource of reserves.Ancient on the books: " the Radix potentillae anserinae all herbal medicine can be treated various hemorrhage and dysentery, and tuber is used as medicine and cures mainly malnutrition, anemia, insufficiency of the spleen diarrhoea, the diseases such as gas loses of suffering from a deficiency of the kidney after being ill ".Put down in writing in the book such as " Chinese medicine voluminous dictionary ", " Chinese herbal medicine is used in Tibet always ": " Radix potentillae anserinae property is put down sweet in the mouth, invigorating spleen and reinforcing stomach, promoting the production of body fluid to quench thirst, QI replenishing and blood tonifying, and both edible can be used as medicine again, cures mainly diseases such as insufficiency of the spleen diarrhoea, malnutrition, anemia; Its all herbal medicine has the effect of the sharp expectorant of astringing to arrest bleeding, cough-relieving.”。And Radix potentillae anserinae also is a kind of nourishing food of Tibetan residential area so far, is not prepared into Chinese medicine patent medicine as yet and occurs in market.
Research shows that Radix potentillae anserinae is rich in 18 seed amino acids and the various trace element of a large amount of starch, fatty acid and needed by human body; Its vitamin C, content of vitamin E are apparently higher than other tuber group food; Have higher medicine and nutritive value, have wide development and utilization prospects.Modern medicine study proof Radix potentillae anserinae has pharmacologically actives such as the immunity of raising, antioxidation, endurance and liver-protecting and blood fat-reducing.
Above-mentioned cognition comes from public publications such as following paper, monograph:
1 Wang Jin, Zhang Jian, Kang Shengli, etc. the Radix potentillae anserinae Study on nutrition is produced in Qinghai. Qinghai medical magazine, 1998,28 (2): 52-53.
2 old Gui are right, Hu Tingjun, and Cheng Fusheng, etc. fern amylose is to mouse lymphocyte propagation and the excretory influence of nitric oxide. Chinese veterinary's science and technology, 2005,35 (9): 735-738.
3 Li Yuan beautiful woman, Yuan Qinsheng. Qinghai-Tibet Radix potentillae anserinae plant Antioxidation Effects. medicine biotechnology, 2004,11 (1): 25-28.
4 old Gui are right, Wang Qin. the extraction of fern amylose and the effect of removing free radical thereof. and Chinese veterinary's science and technology, 2004,34 (4): 59-62.
7 Shang Dejing, Hui Jing, Li Qingwei. Tibet Herba Herminii Analysis of Nutritional and evaluation. Journal of Nutrition, 2002,24 (1): 93-95.
8 Yuan Solenognathus, Xiao Xiaohe, Cai Guangming, etc. the Study on Preparation of liver targeting silverweed cinquefoil essence nanoparticle. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2004,29 (6): 589-590.
9 are new complete, Zhao Yanling, and the beautiful prunus mume (sieb.) sieb.et zucc. in mountain, etc. silverweed cinquefoil essence is to chemical liver injury protective effect Study on Mechanism. PLA's Acta Pharmaceutica Sinica, 2004,20 (4): 259-261.
Summary of the invention
The objective of the invention is to find that the Radix potentillae anserinae extract has significant ischemia resisting effect, can be used for preventing and treating the heart, cerebral ischemia and ischemical reperfusion injury disease, thereby increased the application of Radix potentillae anserinae, and its extract is prepared into Chinese medicine product.
The application of Radix potentillae anserinae extract of the present invention in preparation ischemia resisting medicine is
1, the application of Radix potentillae anserinae extract in the preparation control heart, cerebral ischemia diseases medicine.
2, the application of Radix potentillae anserinae extract in the preparation control heart, cerebral ischemia reperfusion injury medicine.
Ischemia, the damage of ischemia-reperfusion heart and brain are to have a strong impact on one big type of healthy disease of human body always.Along with the change of China's aged tendency of population and life style, important illness, especially ischemical reperfusion injury that this type disease has become the threat people's life are the common complications of interventional therapys such as present clinical cardiovascular and cerebrovascular vessel stenting, Coronary Artery Bypass.In recent years; Chinese scholars has been carried out big quantity research to Chinese herb prevention ischemia, ischemia-reperfusion property heart and brain damage disease; Show that some Chinese medicine has stronger toleration to the body anoxia; Ischemia, ischemical reperfusion injury are had the better protect effect, and Radix potentillae anserinae extract of the present invention has proved that again Chinese medicine has significant effect at control ischemia, ischemia-reperfusion property heart and brain damage disease.
Description of drawings
Accompanying drawing 1 rats in normal control group myocardium myo fibril marshalling (5000 *) electromicroscopic photograph.
Disorderly (5000 *) electromicroscopic photograph of the most of sarcostyle of accompanying drawing 2 ischemia model group rat heart muscles.
The most of sarcostyle of accompanying drawing 3 Radix potentillae anserinae n-butanol portion 0.2g/kg group rat heart muscle is arranged normal (5000 *) electromicroscopic photograph.
The specific embodiment
Radix potentillae anserinae preparation method of extract of the present invention is described below:
Get the dry dried root of Radix potentillae anserinae, pulverize, add 8 times of amount 90% ethanol, refluxed 2 hours at 80 ℃, repeat 2 times, merge extractive liquid, is evaporated to the about 1.40g/ml of proportion, gets alcohol-extracted extract.Extractum is disperseed with 3 times of water gagings, respectively extract 3 times with the petroleum ether, ethyl acetate and the n-butyl alcohol that are equivalent to aqueous dispersion 1/3 volume successively, merge n-butyl alcohol liquid, concentrating under reduced pressure, lyophilization gets n-butanol portion.
Below be the description of test of Radix potentillae anserinae n-butanol portion ischemia resisting effect:
Experiment one
With the scarce effect of the former generation anti-cardiac muscle of myocardial cell anoxia-induced apoptosis model proof Radix potentillae anserinae n-butanol portion
1, method:
(1) rat myocardial cell of former generation is cultivated;
The SD neonatal rat ventricular muscles of getting newborn 1-3d is cut into about 1mm
3The size piece of tissue adds 0.25% trypsin-0.02%EDTA, in 37 ℃ of digestion down.Except that digesting first supernatant discards, to collect each time supernatant and end digestion, centrifugal collection myocardial cell deposition adds the DMEM/F-12 culture medium that contains 10% hyclone, processes cell suspension, in 37 ℃, 5%CO
2Hatch 70min in the incubator, make most non-myocardial cell adherent.The adjustment cell concentration is 1 * 10
5Individual/ml, be inoculated into 24 orifice plates, every hole 1ml; 96 orifice plates, every hole 200 μ l.In the DMEM/F-12 culture medium that contains 10% hyclone, cultivate, changed liquid 1 time in per 2 days.
(2) the myocardial cell anoxia model is set up:
Get the myocardial cell of cultivating 4d, change serum-free DMEM/F-12 culture medium continuous culture 12h after, (charge into 95%N in advance with sugar-free D-Hanks liquid
2-5%CO
2The saturated 30min of gaseous mixture) substitutes the normal cultured base, then rapidly culture plate is moved into and be connected with 95%N
2-5%CO
2In the hypoxia device of gaseous mixture, with oxygen analyser monitoring air vent oxygen concentration (＜1%), 37 ℃ of anoxias are cultivated 6h.
(3) experiment is divided into groups:
Blank group, anoxia model group, anoxia+Radix potentillae anserinae n-butanol portion A group (0.024g/mL), B group (0.012g/mL) and C group (0.006g/mL) totally 5 groups, every group 8 hole are established in experiment.Except that the blank group, other respectively organize equal anoxic treatment 6h, and the normal control group is then in 37 ℃, 5%CO
2Hatch 6h in the incubator synchronously.
(4) anoxia-induced apoptosis myocardial cell MTT experiment:
Take out and respectively organize sample, every hole adds 20 μ l MTT (5g/L), in 37 ℃, 5%CO
2Continue to hatch 4h in the incubator, stop cultivating a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices.Add 150 μ l DMSO jolting 15min, crystallization is fully dissolved,, measure each hole absorbance (O.D) value in measuring wavelength 570nm, reference wavelength 630nm place.
MTT metabolic rate (%)=experimental group O.D value/normal control group O.D value * 100%
(5) biochemical indicator detects:
Get be incubated at 24 orifice plates respectively organize cell culture supernatant, with the activity of colorimetric method for determining LDH, CK, it is active to measure in the cell SOD with xanthine oxidase; With MDA content in the thiobarbituricacid colorimetric method for determining cell, concrete testing process and operational approach are carried out according to the test kit description.
2, result:
In former generation,, myocardial cell anoxia-induced apoptosis model was widely used in the in vitro study of medicine function of resisting myocardial ischemia.When myocardial cell sustained damage because of hypoxic-ischemic, mitochondrial function was unusual, and oxidation-respiration chain is impaired, the electron transport blocking-up, and ATP produces minimizing.Succinate dehydrogenase is one of compound enzyme important in the respiratory chain of succinic acid oxidation, but the dehydrogenation of catalysis succinic acid generates Fumaric acid, generates FADH thereby make FAD accept two hydrogen atoms
2, and then hydrogen passed to C
oQ generates C
oQH
2, the electron transfer process of continuation respiratory chain.So, the activity of succinate dehydrogenase, can the reflecting myocardium cell hypoxia degree of damage.The MTT experimental principle utilizes succinate dehydrogenase can catalysis Thiazolyl blue (MTT) to generate the characteristic of the insoluble coloured product of aubergine exactly, reflects the cytoactive of each group through the mensuration of absorbance.This experimental result shows: 1.00mg/ml, 0.25mg/ml, 0.06mg/ml Radix potentillae anserinae n-butanol portion group; MTT records the O.D value and is significantly higher than model group; Show and all can significantly increase cytoactive (P＜0.01); Explain that the Radix potentillae anserinae n-butanol portion can keep the respiratory function of anoxia-induced apoptosis cell to the infringement of anti-hypoxia to the myocardial cell SDA; When anoxia causes cell membrane damage; LDH, CK will leak outside in the cell; Cause LDH in the culture medium, the active rising of CK; 3 dose groups of this test demonstration are compared LDH in the culture medium, CK activity and are all obviously reduced with model group, show that the Radix potentillae anserinae n-butanol portion can keep the integrity (seeing table 1) of myocardial cell membrane under the acute anoxia condition; SOD has reflected the ability of cell removing free radical; MDA has reflected cell membrane lipid peroxide injury degree; 3 dose groups of this test raise with SOD in model group is compared cell is active; The MDA level produce to descend (seeing table 2), shows that the Radix potentillae anserinae extract can remove the ability of free radical through strengthening myocardial cell, the cell membrane oxidative damage that anti-hypoxia is caused.
Table 1 Radix potentillae anserinae n-butanol portion is to the influence
of anoxia myocardial cell LDH, the outer leakage quantity of CK
*P＜0.05
*Compare with model group P＜0.01
Table 2 Radix potentillae anserinae n-butanol portion is to the influence of anoxia myocardial cell metabolic activity, SOD activity and MDA content
*P＜0.05
*Compare with model group P＜0.01
Experiment two
Adopt pituitrin to cause the effect of mouse cardiac muscle ischemia model proof n-butanol portion resisting myocardial ischemia
1, method:
(1) pituitrin causes the modelling of mouse cardiac muscle ischemic injuries:
90 Kunming kind female mices are divided into 6 groups at random.The high, medium and low dose groups of Radix potentillae anserinae n-butanol portion is irritated the 0.3%CMC-Na solution that stomach gives the Radix potentillae anserinae n-butanol portion respectively, and dosage is respectively 0.3g/kg, 0.15g/kg and 0.075g/kg.Normal group matched group and ischemia model group are all irritated stomach and are given and equivalent 0.3%CMC-Na solution), positive drug FUFANG DANSHEN PIAN solution 0.5g/kg, solvent is a CMC-Na solution, gastric infusion.Respectively organize equal lumbar injection urethane (1g.Kg behind the 60min
-1), treat Animal Anesthesia after, dorsal position is fixed, it is subcutaneous that needle electrode is inserted extremity, every the normal II lead electrocardiogram of mice I is respectively organized in animal electrocardiograph (normal voltage 1mV=10mm, chart speed 50mm/s) record.Ischemia group, positive controls lumbar injection pituitrin 20U/kg, normal group lumbar injection equal volume normal saline.Write down every mice II lead electrocardiogram II.Put to death behind the 30min, get blood and cardiac muscular tissue.
(2) detect index:
Statistics is respectively organized the variation of injected in mice pituitrin front and back electrocardiogram J point.Measure SOD in serum, LDH, CK activity and MDA content.The muscular tissue of coring is processed paraffin section, carries out Nagar-Olsen dyeing, observes the myocardial damage degree.
2, result:
Can cause electrocardiogram to raise during myocardial ischemia or decline, the Radix potentillae anserinae n-butanol portion can cause electrocardiogram J point to change (table 3) to ischemia resisting.The Nagar-Olsen colouring method can be dyed redness with the myocardial damage position, and yellow is then dyed at unmarred position.Coloration result shows; Compare with model group; 0.3g/kg group and the red area that dyes of 0.15g/kg group significantly dwindle; 0.15g/kg the group effect is suitable with 0.5g/kg FUFANG DANSHEN PIAN group, 0.3g/kg group effect is better than 0.5g/kg FUFANG DANSHEN PIAN group, shows that it can effectively dwindle the myocardial damage area that ischemia causes.LDH, CK leak outside and can cause Serum LDH, the active rising of CK during myocardial cell injury, and testing result confirms that the Radix potentillae anserinae n-butanol portion can reduce Serum LDH, CK active (table 4), shows that it can protecting myocardial cell.Can reduce the MDA level, increased SOD active (table 5) shows it and removes the ability of free radical through strengthening myocardial cell, the cell membrane oxidative damage that ischemia resisting is caused.Effect is superior to FUFANG DANSHEN PIAN.
Table 3 Radix potentillae anserinae n-butanol extract causes the influence
of ischemia mice electrocardiogram J point to pituitrin
Annotate: compare a:P＜0.05, b:P＜0.01. with the anoxia model group
Table 4 Radix potentillae anserinae n-butanol portion is to the influence
of myocardial ischemia mice serum LDH, CK level
Annotate: compare with model group,
*: P＜0.05
*: P＜0.01
Table 5 Radix potentillae anserinae n-butanol portion is to the influence
of myocardial ischemia mice serum SOD activity and MDA content
Annotate: compare with model group,
*: P＜0.05
*: P＜0.01
Experiment three
Cause the effect of rat heart muscle ischemia model proof n-butanol portion resisting myocardial ischemia reperfusion injury with the ligation coronary artery
1, method;
Select adult SD rats, body weight 250-300g, be divided at random operative control group, ischemia model group, Radix potentillae anserinae n-butanol portion high (0.2g/kg), in (0.1g/kg), low (0.05g/kg) 3 groups, totally 5 groups, 10 every group.The administration group is irritated stomach and is given the Radix potentillae anserinae extract 1 time every day before experiment, and totally 14 days, operative control group and model group gave the equal-volume solvent.1h after the last administration, the ligation LADCA, treat that rat recovers autonomous respiration after, measure the electrocardiogram of rat, raising rapidly with ECG R wave wave amplitude and ST section is that ligation successfully indicates recording ecg.Open the rat thoracic cavity behind the 30min, cut off ligature, treat that rat recovers autonomous respiration after; Measure the electrocardiogram of rat; With the ECG T wave wave amplitude reduce, the ST section former lifting height 1/2 that descends indicates for pouring into successfully again, puts to death animal after pouring into 120min again, paraffin section is processed by the chamber antetheca cardiac muscular tissue that cores; Carry out Nagar-Olsen dyeing, observe the myocardial damage degree.Observing myocardial ultrastructure through transmission electron microscopy changes.
2, result:
Myocardial ischemia can cause ECG ST section to raise, and experiment shows Radix potentillae anserinae n-butanol portion can raise to the ST section that ischemia resisting causes (seeing table 6).Nagar-Olsen result shows with model and compares that Radix potentillae anserinae n-butanol portion 0.2g/kg and the red area that dyes of 0.1g/kg group rat heart muscle significantly dwindle, and the ratio of normal myocardium tissue increases, and shows that the Radix potentillae anserinae n-butanol portion can alleviate the degree of ischemic myocardium damage.Transmission electron microscope observing confirms that sham operated rats rat heart muscle sarcostyle arranges in order, the clear (see figure 1) of muscle segment, closely arrangement of mitochondrion, regular shape, peplos is complete, ridge is intensive, regular, glycogen granule enriches, intact nuclear membrane, chromatin are even; Behind the ischemia-reperfusion, sarcostyle arrangement disorder (see figure 2), mitochondrion gathering in heaps, swelling.Most of mitochondrion localized membrane is broken, the ridge degraded, and glycogen granule reduces; Nuclear membrane is partly dissolved, and perinuclear space enlarges; Lysosome quantity increases.It is obvious to reach the blood vessel peripheral edema between the fascicula; The most of myocardium myo fibril of Radix potentillae anserinae n-butanol portion 0.2g/kg group rat is arranged basic normal (see figure 3), shows that the Radix potentillae anserinae n-butanol portion can effectively improve the ischemic myocardium ultrastructure.
Table 6 Radix potentillae anserinae n-butanol portion causes the influence that treating myocardial ischemia damage rat ECG ST section changes to CAL
Annotate:
*With model group ratio, p＜0.01.
Experiment four
Effect with former primary cultures of rat neuron cell anoxia-induced apoptosis model proof Radix potentillae anserinae n-butanol portion anti-cerebral ischemia damnification
1, method:
The SD neonatal rat of birth 1-2d is put to death the back and takes out cerebral tissue, separates cerebral cortex; Reject meninges and vascular tissue, be cut into fritter after the flushing, move in the centrifuge tube; Piping and druming makes to organize fully and disperses repeatedly; The centrifugal supernatant of abandoning, the dissolving of DMEM/F12 culture fluid is inoculated in 24 orifice plates that covered with 10 μ g/mL poly-D-lysines in advance, every hole 1ml; 96 orifice plates, every hole 200 μ l.Put 37 ℃ 5% CO
2Cultivate in the incubator.Complete culture solution (DMEM/F12,10% calf serum; Transferrins 10 μ g/mL, 100u/mL penicillin; The 100u/mL streptomycin) keeps; The 1st time full dose is changed liquid, whenever changes liquid 1 time at a distance from 2～3d, half amount later on, and the cytosine arabinoside that when changing liquid on the 3rd day, in culture medium, adds 10 μ g/mL is to suppress neurogliocyte propagation.Full dose is changed liquid behind the 24h.Get the cell of cultivating 8d, change serum-free medium DMEM/F12 continuous culture 24h after, (charge into 95%N2-5%CO in advance with sugar-free D-Hanks liquid
2The saturated 30min of gaseous mixture) substitutes the normal cultured base, then rapidly cultured cell is moved into 95%N of the same race
2-5%CO
2The one-way gas flow hypoxia device that the gaseous mixture balance is crossed, oxygen analyser monitoring air vent oxygen concentration (＜1%) carries out anoxia and cultivates in 37 ℃ of constant incubators.Anoxia is got 24 orifice plates and is respectively organized cell culture supernatant after cultivating 2h, with the activity of colorimetric method for determining LDH, CK, measures SOD activity in the cell with xanthine oxidase; With MDA content in the thiobarbituricacid colorimetric method for determining cell, concrete testing process and operational approach are carried out referring to the test kit description.96 orifice plate cells are with experiment one usefulness mtt assay detection line plastochondria dehydrogenase activity.Experiment be divided into blank group, anoxia model group, Radix potentillae anserinae n-butanol portion high (1.00mg/ml), in (0.25mg/ml), low (0.06mmg/ml) concentration group.Every group 8 hole.
2, result:
1.00mg/ml, 0.25mg/ml, 0.06mg/ml Radix potentillae anserinae n-butanol portion group and model group relatively, the O.D value that mtt assay records enlarges markedly, and shows that the Radix potentillae anserinae n-butanol portion can significantly increase the cell mitochondrial SDA, raising neurocyte vigor; LDH is active in 3 dose groups culture medium reduces, and shows that it has reduced the outer leakage quantity of the LDH that anoxia-induced apoptosis causes; SOD is active in the cell improves, and MDA content reduces, and shows it and removes the ability of free radical through strengthening neuronal cell, the cell membrane oxidative damage that ischemia resisting is caused.(seeing table 7).
Table 7 Radix potentillae anserinae n-butanol portion is to the influence
of anoxia cortex neuron cell mitochondrial dehydrogenase, LDH, SOD activity and MDA level
Annotate: compare with model group,
*P＜0.05
*P＜0.01
Experiment five
Effect with the reperfusion injury of cerebral ischemia reperfusion injury model proof Radix potentillae anserinae n-butyl alcohol anti-cerebral ischemia
1, method:
Get 50 of healthy male SD rats, body weight 250g～300g, be divided at random Sham-operated control group, ischemia model group, Radix potentillae anserinae n-butanol portion high (0.2g/kg), in (0.1g/kg), low (0.1g/kg) concentration group.Totally 5 groups, 10 every group.The administration group is irritated stomach and is given the Radix potentillae anserinae extract 1 time every day before experiment, and totally 14 days, operative control group and model group gave the equal-volume solvent.1h after the last administration, each organizes rat with 10% chloral hydrate 300mg/kg intraperitoneal injection of anesthesia, dorsal position; The cervical region median incision separates ligation bilateral common carotid arteries and vagus nerve and causes the imperfection cerebral ischemia, unclamps ligature behind the 45min; Make blood flow logical again, pour into 90min again.Get blood, measure SOD, LDH activity and MDA content by experiment two methods.Sacrifice of animal, broken end is opened cranium and is peeled off cerebral tissue, inhales the decerebrate surface moisture with filter paper; The weighing botle of packing into takes by weighing weight in wet base, places drying baker to dry to constant weight for 110 ℃ then; Take by weighing dry weight again, calculate brain water content by following formula: brain water content (%)=(weight in wet base-dry weight/weight in wet base) * %
2, result:
3 dose groups of n-butanol portion are compared with model group; All can effectively resist the activity of SOD in serum decline that cerebral ischemic reperfusion in rats causes; LDH activity and MDA level rising (seeing table 8); Show that it can reduce the LDH that cerebral ischemia reperfusion injury causes and leak outside, remove the ability of free radical, the cell membrane oxidative damage that anti-cerebral ischemia is caused through strengthening neurocyte.N-butanol portion 0.2g/kg and 0.1g/kg group brain water content obviously reduce, and with the ischemia model group significant difference are arranged, and show its cerebral edema that causes in the time of can alleviating cerebral ischemia re-pouring (seeing table 9).
Table 8 Radix potentillae anserinae n-butanol portion is to the influence
of cerebral ischemia Serum LDH, SOD activity and MDA level
Annotate: compare with model group,
*P＜0.05
*P＜0.01
Table 9 Radix potentillae anserinae n-butanol portion is to the influence
of cerebral ischemia brain water content
Notes: 1) with matched group ratio, p＜0.05,2) with model group ratio, p＜0.05
More than testing one～experiment three can find out; The Radix potentillae anserinae n-butanol portion has significant resisting myocardial ischemia and ischemical reperfusion injury effect; Experiment four can be found out with experiment five; The Radix potentillae anserinae n-butanol portion has remarkable anti-cerebral ischemia and reperfusion injury and protect, can be used for the preparation control heart, cerebral ischemia and the heart, cerebral ischemia reperfusion injury property disease medicament.Through 1 year animal experimental observation, use medicine of the present invention animal is not found any toxic and side effects.
Below for prepare the embodiment of pharmaceutical preparation with the Radix potentillae anserinae extract:
Embodiment 1, and the Radix potentillae anserinae extract is processed tablet:
Radix potentillae anserinae n-butanol portion 50mg
Starch 25mg
15% starch slurry is an amount of
Magnesium stearate is an amount of
Above-mentioned prescription is processed tablet by traditional processing mode.
Embodiment 2, and the Radix potentillae anserinae extract is processed capsule:
Radix potentillae anserinae n-butanol portion 50mg
Starch 70mg
By traditional pharmaceutical technology preparation, the capsule of packing into No. 2.
Embodiment 3, and the Radix potentillae anserinae extract is processed the oral syrup agent:
Radix potentillae anserinae extract 50mg
Sucrose 200mg
50% ethanol 1ml
Citric acid 5mg
Sodium benzoate 3mg
Water adds to 10ml
Said components added transfer to aequum in the entry, bottling gets final product behind the mixing.
Embodiment 4, and the Radix potentillae anserinae extract is processed granule:
Radix potentillae anserinae extract 50mg
Sucrose 300mg
Fructus Citri tangerinae essence 3mg
Food coloring is an amount of
With processing granule after the said components mixing, packing.
Claims (2)
1. the application of Radix potentillae anserinae extract in the preparation control heart, cerebral ischemia diseases medicine is characterized in that said Radix potentillae anserinae extract is to process with following method: get Radix potentillae anserinae dry root piece, pulverize; Add 8 times of amount 90% ethanol, refluxed 2 hours, repeat 2 times at 80 ℃; Merge extractive liquid, is evaporated to proportion 1.40g/ml, gets alcohol-extracted extract; Extractum is disperseed with 3 times of water gagings, respectively extract 3 times with the petroleum ether, ethyl acetate and the n-butyl alcohol that are equivalent to aqueous dispersion 1/3 volume successively, merge n-butyl alcohol liquid; Concentrating under reduced pressure, lyophilization gets n-butanol portion.
2. the application of Radix potentillae anserinae extract in the preparation control heart, cerebral ischemia reperfusion injury medicine is characterized in that said Radix potentillae anserinae extract is to process with following method: get Radix potentillae anserinae dry root piece, pulverize; Add 8 times of amount 90% ethanol, refluxed 2 hours, repeat 2 times at 80 ℃; Merge extractive liquid, is evaporated to proportion 1.40g/ml, gets alcohol-extracted extract; Extractum is disperseed with 3 times of water gagings, respectively extract 3 times with the petroleum ether, ethyl acetate and the n-butyl alcohol that are equivalent to aqueous dispersion 1/3 volume successively, merge n-butyl alcohol liquid; Concentrating under reduced pressure, lyophilization gets n-butanol portion.
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| CN104958393A (en) * | 2015-06-08 | 2015-10-07 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | Extract of potentilla anserina rhizome and medical application thereof |
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