CN103285115A - Preparation method of active components of compound traditional Chinese medicine for treatment of cardiovascular and cerebrovascular ischemic diseases - Google Patents
Preparation method of active components of compound traditional Chinese medicine for treatment of cardiovascular and cerebrovascular ischemic diseases Download PDFInfo
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Abstract
The invention relates to preparation and identification methods of glycoside active components of a compound traditional Chinese medicine Buyang Huanwu decoction used for treatment of cardiovascular and cerebrovascular ischemic diseases. The compound traditional Chinese medicine is produced by following steps: 60g radix astragali, 9g radix paeoniae rubrathe, 6g ligusticum wallichii, 9g Chinese angelica, 9g lumbricus, 9g flos carthami and 9g peach seed are taken; the ingredients are extracted by water, precipitated by alcohol, eluted by 732 cationic resin, extracted by chloroform and eluted by DA-201 macroporous resin; and then glycoside active components are obtained. The compound traditional Chinese medicine of the invention has effects of tonifying Qi and activating blood, and is used for treatment of cardiovascular and cerebrovascular ischemic diseases. It is confirmed by a large number of clinical trials that the compound traditional Chinese medicine of the invention has actual therapeutic effects on cardiovascular and cerebrovascular ischemic diseases, the effective rate is 90% or more, and simultaneous treatment of principal and subordinate symptoms can be achieved.
Description
Technical field
The present invention relates to a kind of Green Chemistry preparation method of from Chinese medicine compound, extracting medicinal active ingredient, more specifically relate to the preparation method of glycoside active component in the Chinese medicine BUYANG HUANWU TANG.
Background technology
BUYANG HUANWU TANG is one of errors in Medicine Corrected side of Qing Dynasty's Wang Qingren, is made up of the Radix Astragali, Radix Angelicae Sinensis, Radix Paeoniae Rubra, Pheretima, Rhizoma Chuanxiong, Semen Persicae, Flos Carthami, and its effect is benefiting qi and nourishing blood, promoting blood circulation to remove obstruction in the collateral; Modern Chinese medicine is clinical to be used for the treatment of the cardiovascular and cerebrovascular vessel ischemic diseases more, has effects such as antithrombotic, blood fat reducing, the neural reparation of promotion and raising immunologic function.
BUYANG HUANWU TANG is the classic prescriptions of Chinese traditional treatment ischemic cardio cerebrovascular diseases, ours studies show that in recent years, glycoside active component among this side is the main medicinal active ingredient of its treatment cardiovascular and cerebrovascular vessel ischemic diseases, the glycoside component has the effect of anti-cerebral ischemia, anti-artery thrombosis and anti-angiogenic neointimal hyperplasia, is the main pharmacological component of this side's cardiovascular and cerebrovascular vessel effect.
Though Chinese patent medicine and the Western medicine of multiple treatment cardiovascular and cerebrovascular vessel ischemic diseases are arranged on city's field boundary at present, but side effect is more or less arranged, following side effect can occur as the Western medicine for the treatment of cardiovascular and cerebrovascular vessel ischemic diseases: 1. central nervous system: depression, delirium; 2. peripheral nervous system: lower limb severe pain, spasmodic torticollis; 3. digestive system: intractable singultus, diarrhoea, peptic ulcer; 4. allergy: systemic rash, fixed pattern drug eruption, symmetry erythra etc.; 5. blood system: leukopenia, hemorrhage etc.The Chinese patent medicine for the treatment of ischemic cardio cerebrovascular diseases commonly used is many to be made up of blood-activating and stasis-removing, as: injections such as compound Salviae Miltiorrhizae pin, Radix Salviae Miltiorrhizae Injection, ligustrazine pin, MAILUONING pin, QINGKAILING pin, XINGNAOJING pin, breviscapine pin, thromboembolism cleansing pin, Ahylysantinfarctase pin; Or oral agents such as NIAOXUEKANG KOUFUYE, QINGKAILING KOUFUYE, Radix Salviae Miltiorrhizae Tabellae, Annaowan bolus, the strong refreshing pellet of refreshment, NAOXUESHUAN PIAN, NAOAN JIAONANG, TONGXINLUO JIAONANG, HUATUO ZAIZAO WAN, apoplexy return of spring sheet, XUESHUANXINMAINING, Piracetam Tablet and ginkgo agent, the effect of blood circulation promoting and blood stasis dispelling is arranged mostly, this series products has certain curative effect, but there is the shortcoming of effect instability, poor repeatability in this series products.
Because chemical composition of Chinese materia medica complexity, the extract of the extraction process by water preparation that Chinese medicine adopts, though contain the medicinal active ingredient of main treatment cardiovascular and cerebrovascular vessel ischemic diseases, but also mix inactive impurity compositions such as amounts of protein, polysaccharide, quintessence oil, tannin, inorganic salt and organic acid are arranged, there are defectives such as active component purity is not high, the content of drug effect components of resisting cardiovascular ischemic diseases is lower, cause the effect of medicine not strong, the quality of product, scientific and technological gold content, added value are all not high.
Summary of the invention
The objective of the invention is to solve in the BUYANG HUANWU TANG defectives such as active component purity content of drug effect components not high, the resisting cardiovascular ischemic diseases is lower, so a kind of preparation technology who prepares the Chinese medicine compound BUYANG HUANWU TANG glycoside active component of the treatment cardiovascular and cerebrovascular vessel ischemic diseases simple, that active constituent content is high, cheap is provided.
The preparation technology 1 of active component:
Get each medical material by Radix Astragali 4-6 part, Radix Paeoniae Rubra 1-3 part, Rhizoma Chuanxiong 1-2 part, Radix Angelicae Sinensis 1-3 part, Pheretima 1-3 part, Flos Carthami 1-3 part, Semen Persicae 1-3 part weight ratio, soak 60min, reflux extracts twice, and amount of water is respectively 8 times, 6 times, and extraction time is respectively 60min and 40min.Filter, merge filtrate twice, with 70 ℃ of vacuum concentration of filtrate to 1g/mL(by crude drug); Add ethanol and transfer to and contain determining alcohol 85%, get water alcohol liquid, place precipitation, sucking filtration after 24 hours, 55 ℃ of vacuum of pure liquid reclaim ethanol and evaporate to dryness water to 1g/mL(by crude drug); Repeated treatments, precipitate 3 times, the last concentration of alcohol of transferring reaches 90%, being placed to deposit-free generates again, sucking filtration, filtrate recycling ethanol, and evaporate to dryness to 1g/mL(by crude drug), must contain glycosides, aglycon, alkaloid, the sample liquid of aminoacid blending ingredients 1., 1. sample liquid transfers pH is 2, quick 732 type cation exchange resin columns by having made the transition and handled well with 7% hydrochloric acid in advance, use the distilled water eluting, collect the alpha-Naphthol concentrated sulphuric acid, the acetic anhydride concentrated sulphuric acid, the hydrochloric acid magnesium powder, be the eluent that reactions such as green fluorescence are positive under the aluminum chloride methanol solution ultraviolet light, must contain glycosides, aglycon blending ingredients sample liquid 2., sample liquid is 2. behind 70 ℃ of vacuum concentration, it is neutral transferring pH with ammonia, uses concentrated solution and chloroform (1:1) amount to carry out the gradation continuous extraction then, reclaims the chloroform evaporate to dryness; Collecting the alpha-Naphthol concentrated sulphuric acid is positive, be yellow-green fluorescence respectively under acetic anhydride concentrated sulphuric acid, hydrochloric acid magnesium powder, the aluminum chloride methanol solution ultraviolet light simultaneously, the water liquid that vanillin concentrated sulphuric acid silica gel plate drop reaction is the positive reaction of purple gets glycoside component crude samples liquid, the refining sugar that removes of glycoside component crude samples liquid: get the DA-201 macroporous resin, in advance with the 95% soak with ethanol previous cleaning that spends the night, use 3% salt acid elution then, wash with water to neutrality; With the washing of 7% sodium hydroxide, wash with water to neutrality then, be soaked in water, the dress post.Add glycosides crude samples liquid on the DA-201 macroporous resin column; Washing carbohydrate content to aforementioned aglycon reaction earlier with water has just occurred till the positive reaction; Wash by the glycoside component of macroporous resin adsorption with 60% ethanol then, reclaim ethanol and get glycoside component extractum, 60 ℃ of vacuum dryings get the glycoside component.
The preparation technology 2 of active component:
Get each medical material by 20 parts of the Radixs Astragali, 3 parts of Radix Paeoniae Rubra, 2 parts of Rhizoma Chuanxiongs, 3 parts of Radix Angelicae Sinensis, 3 parts of Pheretimas, 3 parts on Flos Carthami, 3 parts of weight ratios of Semen Persicae, soak 60min, reflux extracts twice, and amount of water is respectively 8 times, 6 times, and extraction time is respectively 60min and 40min; Filter, merge filtrate twice, with 70 ℃ of vacuum concentration of filtrate to 1g/mL(by crude drug); Add ethanol and transfer to and contain determining alcohol 85%, get water alcohol liquid, place precipitation, sucking filtration after 24 hours, 55 ℃ of vacuum of pure liquid reclaim ethanol and evaporate to dryness water to 1g/mL(by crude drug); Repeated treatments, precipitate 3 times, the last concentration of alcohol of transferring reaches 90%, being placed to deposit-free generates again, sucking filtration, filtrate recycling ethanol, and evaporate to dryness to 1g/mL(by crude drug), must contain glycosides, aglycon, alkaloid, the sample liquid of aminoacid blending ingredients 1., 1. sample liquid transfers pH is 2, quick 732 type cation exchange resin columns by having made the transition and handled well with 7% hydrochloric acid in advance, use the distilled water eluting, collect the alpha-Naphthol concentrated sulphuric acid, the acetic anhydride concentrated sulphuric acid, the hydrochloric acid magnesium powder, be the eluent that reactions such as green fluorescence are positive under the aluminum chloride methanol solution ultraviolet light, must contain glycosides, aglycon blending ingredients sample liquid 2., sample liquid is 2. behind 70 ℃ of vacuum concentration, it is neutral transferring pH with ammonia, uses concentrated solution and chloroform (1:1) amount to carry out the gradation continuous extraction then, reclaims the chloroform evaporate to dryness; Collecting the alpha-Naphthol concentrated sulphuric acid is positive, be yellow-green fluorescence respectively under acetic anhydride concentrated sulphuric acid, hydrochloric acid magnesium powder, the aluminum chloride methanol solution ultraviolet light simultaneously, the water liquid that vanillin concentrated sulphuric acid silica gel plate drop reaction is the positive reaction of purple gets glycoside component crude samples liquid, the refining sugar that removes of glycoside component crude samples liquid: get the DA-201 macroporous resin, in advance with the 95% soak with ethanol previous cleaning that spends the night, use 3% salt acid elution then, wash with water to neutrality; With the washing of 7% sodium hydroxide, wash with water to neutrality then, be soaked in water, the dress post.Add glycosides crude samples liquid on the DA-201 macroporous resin column.Washing carbohydrate content to aforementioned aglycon reaction earlier with water has just occurred till the positive reaction; Wash by the glycoside component of macroporous resin adsorption with 60% ethanol then, reclaim ethanol and get glycoside component extractum, 60 ℃ of vacuum dryings get the glycoside component.
The qualitative and quantitative analysis of glycoside active component
Mainly contain astragaloside, peoniflorin, amygdaloside etc. in the BUYANG HUANWU TANG in the glycoside component, carry out corresponding qualitative and quantitative analysis with the active constituents of medicine that mainly exists in its glycoside component in this invention.
1. chemistry is differentiated
Get glycoside component dissolve with ethanol and make 10mgmL respectively
-1Sample liquid, carry out alpha-Naphthol concentrated sulphuric acid, bismuth potassium iodide, silico-tungstic acid, iodine-potassium iodide, acetic anhydride concentrated sulphuric acid, hydrochloric acid magnesium powder, alkaline picric acid, the potassium ferricyanide---reactions such as ferric chloride respectively, the results are shown in table 1.
Table 1 BUYANG HUANWU TANG glycoside active component chemistry is differentiated the phenomenon result
| Chemical test | The result |
| The dense H2SO4 of alpha-Naphthol | ++ the purple ring |
| Bismuth potassium iodide | — |
| Silico-tungstic acid | — |
| Iodine-potassium iodide | — |
| The dense H2SO4 of acetic anhydride | ++ be yellow to green color |
| The hydrochloric acid magnesium powder | ± redness or aubergine |
| Alkaline picric acid | + red or purplish red, orange red |
| The potassium ferricyanide-ferric chloride | ± |
Annotate: ++ positive findings is very obvious; + positive findings is obvious; ± positive findings has interference;-positive findings is not obvious.
2. the assay of effective active composition in the glycoside component
2.1 the mensuration of Astragaloside content:
Chromatographic condition
Chromatographic column: C18 post 4.6 * 250mm, 5 μ m; Detector: evaporat light scattering, 60 ℃ of temperature, gas flow 1.5L/min, gain 1.0; Mobile phase: methanol (80/20, V:V).Flow velocity: 1mLmin
-1Column temperature: 30 ℃.
Astragaloside reference substance liquid: precision takes by weighing astragaloside reference substance 9.1mg,, makes and contains astragaloside 9.1 mgmL to 10mL with methanol constant volume
-1Reference substance liquid; Each sample introduction 10 μ L.
Sample treatment: take by weighing glycoside component sample 50.9mg, use the 50mL dissolve with ethanol, filter, evaporate to dryness in the water-bath, with 10mL methanol quantitatively to scale namely; Each sample introduction 10 μ L.
2.2 peoniflorin, amygdaloside Determination on content:
Chromatographic condition
Chromatographic column: C18 post 4.6 * 250mm, 5 μ m; Detect wavelength: 218nm.Mobile phase: methanol (27/73, V:V); Flow velocity: 1mLmin
-1Column temperature: 30 ℃.
Peoniflorin reference substance liquid: precision takes by weighing peoniflorin reference substance 7.7mg,, makes and contains peoniflorin 0.77 mgmL to 10mL with methanol constant volume
-1Reference substance liquid; Each sample introduction 10 μ L.
Amygdaloside reference substance liquid: precision takes by weighing amygdaloside reference substance 8.2mg,, makes and contains amygdaloside 0.82 mgmL to 10mL with methanol constant volume
-1Reference substance liquid; Each sample introduction 10 μ L.
Sample treatment: get glycoside component sample 50.9mg, use the 50mL dissolve with ethanol, filter, evaporate to dryness in the water-bath, with 10mL methanol quantitatively to scale namely; Each sample introduction 10 μ L.
Its above-mentioned three kinds of materials are carried out after high performance liquid chromatography (HPLC) detects, with its peak area by with relevant three kinds of standard of physical curves conversion, calculate astragaloside, peoniflorin, amygdaloside content in its glycoside active component.
The glycoside component is correlated with finds that its content increases significantly after the qualitative and quantitative detection, reached its goal of the invention, overcome the deficiency of traditional product and technology, solve defectives such as medicinal active ingredient purity content of drug effect components not high, the resisting cardiovascular ischemic diseases is lower, the extraction, separation, the purification that are suitable for BUYANG HUANWU TANG glycoside active component are for preparation BUYANG HUANWU TANG glycoside active component related preparations in next step commercial production provides method.
The animal pharmacodynamics evaluation experimental of glycoside active component in the BUYANG HUANWU TANG
(1) research of BUYANG HUANWU TANG glycoside active component Chinese People's Anti-Japanese Military and Political College Mus cerebral ischemia effect
Get the BUYANG HUANWU TANG glycoside active component that this extraction process is extracted, standby with degerming behind the dissolved in distilled water; Nimotop vial is the Zhejiang limited branch of one new pharmacy share department product, lot number 01092001; Glycoside active component and nimodipine are carried out controlled trial, method is: get 24 cleaning level SD rat (male and female half and half, body weight 305.0 ± 14.5g) is divided into by the completely random method: Sham-operated control group (5), model control group (8), glycoside active component group (6) and nimodipine group (5); Administering mode is: the equal lumbar injection equivalent of Sham-operated control group and model control group normal saline (10mL/kg), nimodipine group lumbar injection nimodipine (0.002g/kg), glycoside active component group injection glycoside active component (0.336g/kg); Administration adopts left side internal carotid artery line bolt legal system to be equipped with the inaccessible focal cerebral ischemia model of intraluminal middle cerebral artery occlusion in rats immediately after 5 minutes; Body temperature is monitored by anus temperature meter in the art, and keeps the anus temperature at 36.5-37 ℃ with the electric filament lamp heating, prevents that low temperature is to the influence of cerebral ischemia.After 6 hours, adopt blind method assessment neurological deficits score in cerebral ischemia; Put to death animal then, get cerebral tissue rapidly, after normal saline flushing is clean, cerebral tissue is cut into the thick crown section of 2mm, put into 2%2 rapidly, 3, in 5-RT (TTC) solution, 37 ℃ of warm the region between the heart and the diaphragm 30min dyeing, 4% formalin is fixed 24 hours, take pictures with digital camera, measure infarction kitchen range area (rose-red zone is normal cerebral tissue, and pale district is the infarction cerebral tissue) with the color pathological image analyzer, each brain sheet infarct size * thickness is total Infarction volume, analyze as relative cerebral infarction volume with the long-pending percentage rate that accounts for full brain volume of infarction stove, the results are shown in Table 2.
Table 2 respectively organize cerebral infarction volume and function of nervous system behavior scoring relatively (
S)
The relative cerebral infarction volume of group number of animals (%) neurological deficits score
Model group 8 22.14
5.67 △ △ 9.2
0.5 △ △
Glycosides group 6 10.98
3.84 △ △ * * 4.8
1.2 △ △ * *
Nimodipine group 5 9.08
4.01 △ * * 4.8
1.9 △ *
Sham operated rats rat function of nervous system sign is normal, and neurological deficits score is 0, has not seen the infarction kitchen range, and the cerebral infarction volume is 0; Model group neurological deficit sign is obvious, and neurological deficits score is significantly higher than sham operated rats (p＜0.01), and the cerebral infarction volume is significantly higher than sham operated rats (p＜0.01) relatively; Glycoside active component group and nimodipine group rat neurological deficit sign are obviously improved, the neurological deficit scoring all significantly is lower than model group (p＜0.01 and p＜0.05), and the cerebral infarction volume also significantly is lower than model group (p＜0.01 and p＜0.05) relatively; Two groups of rat neurological deficits score and relative cerebral infarction volume basically identical, difference do not have significance (p〉0.05); Show the glycoside active component of BUYANG HUANWU TANG and the effect that nimodipine all has anti-cerebral ischemia.
(2)
The research of the anti-artery thrombosis effect of BUYANG HUANWU TANG glycoside active component
1. mouse anti thrombosis experiment: get the BUYANG HUANWU TANG glycoside active component that this extraction process is extracted, standby with degerming behind the dissolved in distilled water; Ticlid (Ticlopidine) is available from Hangzhou Sano-Synth labo people's livelihood pharmaceutical Co. Ltd, 0.25g/ sheet, lot number: 051; Low molecular heparin (LMWH) is for Jiangsu Jiangshan Pharmaceutical Co provides, 5000 anti-X aIU/ml, lot number: 20040602; With glycoside active component and LMWH, Ticlid carries out control experiment, method is: cleaning level Kunming kind white mice, male and female half and half, body weight 18-22g, be divided into by the completely random method: model group (intraperitoneal injection of saline 10mL/kg), glycosides height (lumbar injection glycoside active component 0.5g/kg), in (lumbar injection glycoside active component 0.25g/kg), low (lumbar injection glycoside active component 0.125g/kg) dosage group, Ticlid group (irritating stomach Ticlid 0.05mg/kg), LMWH organizes (subcutaneous injection LMWH750u/kg), administration, the mouse back of modeling simultaneously subcutaneous injection 1% carrageenin 50mg/kg, the same administration in 12 hours is 1 time behind the injection carrageenin, behind the injection carrageenin, measure arteria caudalis thrombosis length in 24 hours, the results are shown in Table 3.
Annotate: compare with model group
△P＜0.05
Model group can be seen tangible arteria caudalis thrombosis; Compare with model group, glycosides high dose group arteria caudalis thrombosis length significantly reduces (p＜0.05), though and dosage group and low dose group arteria caudalis thrombosis length are lower than model group but difference does not have significance (p〉0.05) in the glycosides, there is certain dose-effect relationship in the anti thrombotic action of the high, medium and low dosage of glycosides; The thrombosis length of LMWH group and ticlid see ticlopidine group also all is lower than model group (p＜0.05); Showing that glycoside active component and LMWH, ticlid see ticlopidine are the same all has an antithrombotic effect.
2. rat anti thrombosis experiment: cleaning level SD rat, male and female half and half, body weight 225.0
25.0g, be divided into by the completely random method: Sham-operated control group (intraperitoneal injection of saline 10mL/kg), model group (intraperitoneal injection of saline 10mL/kg), glycoside active component group (lumbar injection glycoside active component 1.63g/kg), Low molecular heparin (LMWH) group (subcutaneous injection Low molecular heparin 1150u/kg), Ticlid group (irritating stomach Ticlid 0.03g/kg), the same administration is 1 time after 12 hours and 24 hours; After the administration 15 minutes with the chloral hydrate intraperitoneal injection of anesthesia; separate bilateral carotid, length is 2cm, at left common carotid artery underlying small pieces plastic sheeting (4cm * 1.8cm); for the protection of the blood vessel surrounding tissue, other each group has suction the 30%FeCl of 20 μ L except sham operated rats
3The small pieces filter paper of solution (1cm * 1cm) spread on the left carotid, sham operated rats is applied equivalent normal saline filter paper, 30 minutes removal scraps of paper behind the deposited scraps of paper, remove behind the scraps of paper and to cut left carotid thrombosis position blood vessel in 90 minutes, blot residual blood, measure its weight with electronic analytical balance, the results are shown in Table 4.
Table 4 respectively organize thrombus weight relatively (
S)
| Group | Dosage | The Mus number | Thrombus weight (mg) |
| Sham operated rats | --- | 12 | 0.0
 |
| Model group | 0 | 12 | 10.1
 |
| The glycosides group | 1.63g/kg | 12 | 7.1
 |
| The Low molecular heparin group | 1150u/kg | 12 | 7.8
 |
| The Ticlid group | 0.03g./kg | 12 | 7.9
 |
Annotate: with sham operated rats ratio ▲ P＜0.05, ▲ ▲ P＜0.01.With model group Bi ﹡ P＜0.05.
The Sham-operated control group left carotid does not have thrombosis and forms; The visible occluding thrombus shape of model group left common carotid artery face.Compare with model group, glycoside active component group, LMWH group and Ticlid group thrombus weight all significantly are lower than model group (P＜0.05); Show that glycoside active component and low molecular weight heparin, Ticlid are the same, can suppress the thrombosis due to the ferric chloride injured blood vessel.
(3) research of BUYANG HUANWU TANG glycoside active component Chinese People's Anti-Japanese Military and Political College Mus vascellum endometrial hyperplasia effect
Get the BUYANG HUANWU TANG glycoside active component that this extraction process is extracted, standby with degerming behind the dissolved in distilled water; Atorvastatin, the trade name lipitor, specification 10mg/ sheet, Godecke GmbH produces, pfizer inc packing, lot number: 65837019; Cleaning level SD rat, body weight: 300-350g, male, be divided into by the completely random method: sham operated rats, model group, atorvastatin group and glycoside active component group; Rat with 10 ﹪ chloral hydrate intraperitoneal injection of anesthesia after, make the cervical region median incision, expose and free left common carotid artery 1～1.5cm, the distal end ligation, after the left common carotid artery proximal part is with the bulldog clamp blocking-up, cut one " V " shape otch in the common carotid artery distal end, insert the 2.0F foley's tube, careful operation conduit is crossed aortic arch and is gone into breast, and down to ventral aorta, the degree of depth is about 6～7cm, inject the full sacculus of normal saline 0.1～0.2mL from the conduit tail end, make inflation that significantly resistance sense be arranged when pulling back, keep the resistance unanimity slowly to pull back conduit to the aortic arch place, so repeat 6 times, be 180 ° of axis rotation sacculus again with the seal wire, make sacculus go to the another side of lumen of vessels, repeat again 6 times according to last method, fully strip off endothelium.Withdraw from conduit after finishing, ligation left common carotid artery proximal part blood vessel is with hemostasis, layer-by-layer suture muscle, hypodermic layer and skin layer; Postoperative lumbar injection penicillin 200,000 units are used in conjunction 3 days; After modeling the 1st day, sham operated rats and model group were irritated stomach equivalent distilled water; The glycosides group is irritated stomach glycoside active component (7.1g/Kg); The atorvastatin group is irritated stomach atorvastatin (10mg/Kg), and be administered once every day, continuous irrigation stomach 14 days; Animal is put to death in the 15th day anesthesia back after modeling, take out damage section thoracic aorta, get blood vessel 1cm in the endothelium section of stripping off thoracic aorta same area, 4% paraformaldehyde is fixed 24 hours, the dehydration of ethanol gradient, paraffin embedding, after every section blood vessel is interrupted 8～10 of uniformly slicings, Masson method trichrome stain, respectively get 3 at random carries out om observation to every rat specimen, take a picture, carry out the vascular morphology measuration, with MIAS medical image analysis system measurement and calculate inner membrance area (IA), inner film thickness (IT), inner membrance area hypertrophy ratio (HRIA), inner film thickness hypertrophy ratio (HRIT), inner membrance/media area is than (IA/MA), inner membrance/media thickness the results are shown in Table 5 than (IT/MT).
Table 5 respectively organize vascular morphology learn measuring index relatively (
± s)
| HRIA(％) | HRIT(％) | IA/MA | IT/MT |
| 0±0 | 0±0 | 0.00±0.00 | 0.00±0.00 |
| 27±9△△ | 29±10△△ | 0.40±0.18△△ | 0.42±0.19△△ |
| 18±5△△☆☆ | 18±5△△☆☆ | 0.21±0.07△△☆☆ | 0.22±0.07△△☆☆ |
| 14±5△△☆☆ | 14±5△△☆☆ | 0.16±0.06△△☆☆ | 0.17±0.06△△☆☆ |
Compare △ △ P＜0.01 with sham operated rats; Compare ☆ P＜0.05, ☆ ☆ P＜0.01 with model group.
The sham operated rats aortic tunica intima does not have hypertrophy; The visible significantly hypertrophy of model group aortic tunica intima, luminal stenosis, the vascellum endometrial hyperplasia index all is significantly higher than sham operated rats (P＜0.01); Compare with model group, atorvastatin group and glycoside active component group vascellum endometrial hyperplasia index all significantly reduce (P＜0.01); Show the same equal hypertrophy that can significantly suppress tunica intima behind the vascular endothelial injury with atorvastatin of the glycoside active component of BUYANG HUANWU TANG.
Claims (8)
1. the preparation method of the total glycoside active component of BUYANG HUANWU TANG, it is characterized in that: the active component that relates among the present invention is to extract to form from the Chinese medicinal formulae BUYANG HUANWU TANG.
2. the preparation method of the total glycoside active component of BUYANG HUANWU TANG according to claim 1 is characterized in that: write out a prescription and be made up of the Radix Astragali, Radix Paeoniae Rubra, Rhizoma Chuanxiong, Radix Angelicae Sinensis, Pheretima, Flos Carthami, Semen Persicae.
3. the preparation method of the total glycoside active component of BUYANG HUANWU TANG according to claim 2, each component preferred weight consists of in the Chinese medicine decoction: Radix Astragali 4-6 part, Radix Paeoniae Rubra 1-3 part, Rhizoma Chuanxiong 1-2 part, Radix Angelicae Sinensis 1-3 part, Pheretima 1-3 part, Flos Carthami 1-3 part, Semen Persicae 1-3 part.
4. the preparation method of the total glycoside active component of BUYANG HUANWU TANG according to claim 2, the optimum weight of each component consists of in the Chinese medicine decoction: 20 parts of the Radixs Astragali, 3 parts of Radix Paeoniae Rubra, 2 parts of Rhizoma Chuanxiongs, 3 parts of Radix Angelicae Sinensis, 3 parts of Pheretimas, 3 parts on Flos Carthami, 3 parts in Semen Persicae.
5. the preparation method of glycoside active component in the BUYANG HUANWU TANG according to claim 3, it is characterized in that: described separate mode is through water extract-alcohol precipitation, obtains by 732 cationic resin, chloroform extraction and DA-201 macroporous resin eluting.
6. according to the preparation method of glycoside active component in the described BUYANG HUANWU TANG of claim 1-5, mainly comprise astragaloside, peoniflorin, amygdaloside etc. in the active component of gained.
7. the preparation method of total glycoside material in the BUYANG HUANWU TANG according to claim 6, the active component of its preparation can be made into any pharmaceutically acceptable dosage form.
8. the preparation method of total glycoside material in the BUYANG HUANWU TANG according to claim 6, the preferred dosage form of the active component of its preparation is: dosage forms such as capsule, granule, tablet, oral liquid, syrup, microcapsule, powder, drop pill or pill.
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| CN104147034A (en) * | 2014-08-15 | 2014-11-19 | 湖南中医药大学 | Traditional Chinese medicine composition used for inhibiting propagation of vascular smooth muscle cells, and preparation method and application thereof |
| CN104887757A (en) * | 2015-06-16 | 2015-09-09 | 圣原健康产业有限公司 | Pulse-invigorating and heart-nourishing traditional Chinese medicine composition and massage cream |
| CN106176905A (en) * | 2016-08-29 | 2016-12-07 | 湖南中医药大学 | A kind of Radix Astragali and the application in treatment of vascular neointimal hyperplasia of the angelica medicament compositions |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104147034A (en) * | 2014-08-15 | 2014-11-19 | 湖南中医药大学 | Traditional Chinese medicine composition used for inhibiting propagation of vascular smooth muscle cells, and preparation method and application thereof |
| CN104887757A (en) * | 2015-06-16 | 2015-09-09 | 圣原健康产业有限公司 | Pulse-invigorating and heart-nourishing traditional Chinese medicine composition and massage cream |
| CN104887757B (en) * | 2015-06-16 | 2019-04-02 | 圣原健康产业有限公司 | The Chinese medicine composition and massage cream of freeing vessels and nourishing heart |
| CN106176905A (en) * | 2016-08-29 | 2016-12-07 | 湖南中医药大学 | A kind of Radix Astragali and the application in treatment of vascular neointimal hyperplasia of the angelica medicament compositions |
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