CN1476891A - Chinese medicine composition for curing apoplexy and its preparation method - Google Patents
Chinese medicine composition for curing apoplexy and its preparation method Download PDFInfo
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Abstract
The present invention discloses a Chinese medicine composition for curing apoplexy, its preparation method and quality contrl method. The composition mainly is formed from (by weight portion) 150-220 portions of gardenia fruit and 3-9 portions of notoginseng total saponin, and its preparation method includes the following steps: using ethyl alcohol to make the gardenia fruit undergo the process of diacolation treatment, concentrating to obtain the extract, making said extract pass through the active carbon column to obtain gardenia fruit semifinished product and using proper quantity of injection water to dissolve said semifinished product; dissolving notoginseng total saponin in proper quantity of injection water, filtering, and uniformly mixing notoginseng liquid medicine and gardenia fruit liquid medicine, regulating pH value, filtering with filter membrane, not-pressing and sterilizing.
Description
Invention field
The present invention relates to a kind of Chinese medicine composition, particularly be used for the treatment of the Chinese medicine composition of apoplexy, relate to the preparation method and the method for quality control of said composition simultaneously.
Background technology
Apoplexy belongs to modern medicine cerebrovascular category, is a kind of disease with higher incidence, case fatality rate and disability rate, be the commonly encountered diseases of middle-aged and elderly people, and age of onset has the trend of rejuvenation.In apoplexy, the ratio of cerebral infarction is higher, and according to Epidemiological study, cerebrovascular 18% is a cerebral hemorrhage, and 82% is cerebral infarction.Relevant survey data shows that the annual apoplexy sickness rate of China is 115.8/10 ten thousand, and particularly dysnoesia behind the apoplexy and limbs are disabled, and have become serious social concern and difficult medical problem; Morbidity back in the patient of first aid survival, has 73%-86% hemiplegia to occur in 1 week, and 71%-77% has moving difficulty, 47% sitting alone, and 44% has the somesthetic sensibility obstacle, and the disability rate of apoplexy is very high.It is generally acknowledged that acute stage alleviates the scope of brain tissue damage and degree is to reduce the important step of disability rate.
The clinical treatment of cerebral infarction, the traditional Chinese medical science mainly are to use square medicines such as suppressing the hyperactive liver to relieve the wind syndrome, eleminating phlegm and freeing channels, promoting flow of QI and blood, the refreshment of having one's ideas straightened out at present.Because the restriction of aspects such as route of administration is difficult to bring into play the treatment advantage in acute stage.Even some injections such as MAILUONING, QINGKAILING etc. are arranged, but because its complicated component, mechanism of action is not clear and definite as yet, and the purposiveness of clinical practice is relatively poor; Function of promoting blood circulation to disperse blood clots such as XUESHUANTONG (XUESAITONG) injection are better, but very little for causing the obstructed factor affecting of brain network stasis of blood resistance, clinical efficacy does not obtain to significantly improve.Doctor trained in Western medicine still has many insoluble problems according to the medicine and the therapeutic scheme of cerebral ischemia mechanism result of study initiative in recent years.For example, think the medicine nmda receptor antagonist (MK-801) that can stop the ischemia damage process, calcium-ion channel antagonists (nimodipine) etc. at present, can reduce the scope of ischemic pathological changes, but with making any distinction between the neurotransmission of blocking-up EAA mediation, influence some normal physiological functions, comprise and repair necessary neural plasticity, may bring adverse effect for the prognosis rehabilitation.The ischemic cerebral edema of acute stage is important pathological process of apoplexy.Because the mixing of cytotoxicity and angiogenic two class factors, the former is at the depletion of film ionic pump, and treatment should take to reverse energy exhaustion promptly provides oxygenated blood to recover pumping function, and at this moment, the high osmotic agent effect is limited; The ischemia angiogenic edema that blood brain barrier collapse produces after a few hours, high osmotic agent should prove effective according to reason, however that CT scan is observed high osmotic agent is not remarkable to cerebral infarction swelling effect, and mannitol has the side effect of concurrent edema knock-on; As if the still difficult appraisal of dexamethasone curative effect on probation does not have benifit even steroid hormone is widely applied yet, even may be harmful to ischemic neuron; Glycerol has certain effect in the cerebrovascular survivor, but causes diabetes control difficulty.
Therefore, a kind of ischemia cascade reaction of preventing of clinical needs damages, and improves the slight irrigation stream mode of cerebral ischemia, thus effective neuroprotective cell, the relative clearly modern Chinese medicine of effective substance compound injection with the mechanism of action.
Summary of the invention
One object of the present invention is to disclose a kind of Chinese medicine composition of new treatment apoplexy; Another object of the present invention is the method for the Chinese medicine composition of a kind of new treatment apoplexy of open preparation; The object of the invention also is to disclose a kind of method of quality control of new Chinese medicine composition.The crude drug of pharmaceutical composition of the present invention is formed and proportioning following (by weight): the A scheme:
Fructus Gardeniae 150-220 weight portion Radix Notoginseng 70-110 weight portion.Preferably:
Fructus Gardeniae 170-200 weight portion Radix Notoginseng 80-100 weight portion.The B scheme
Fructus Gardeniae 150-220 weight portion Radix Notoginseng total arasaponins 3-9 weight portion is preferred:
Fructus Gardeniae 170-200 weight portion Radix Notoginseng total arasaponins 5-7 weight portion C scheme
Jasminoidin 3.8-11.5 weight portion Radix Notoginseng total arasaponins 3-9 weight portion.Preferably:
Jasminoidin 6.3-8.9 weight portion Radix Notoginseng total arasaponins 5-7 weight portion.
Press practice of pharmacy, the invention described above preparation of pharmaceutical compositions can be become the various clinical pharmaceutical dosage form, comprise the dosage form of oral formulations or parenterai administration.Said oral formulations is selected from a kind of in the middle of the tablet, capsule, pill, granule, suspensoid, drop pill, oral liquid; Said parenterai administration dosage form is selected from a kind of in the middle of injection, aerosol, suppository or the subcutaneous administration dosage form.
Medicine of the present invention also can add conventional drug excipient, as solvent, disintegrating agent, correctives, antiseptic, coloring agent etc.
The preparation method that the B scheme is made injection is:
Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, with the 65-80% ethanol percolation that 7-9 doubly measures, collect percolate, reclaim ethanol, the clear paste that to be concentrated into 55-70 ℃ of relative density be 1.00-1.20, add the dilution of 3-5 times of water gaging, stir evenly, 0-4 ℃ cold preservation 45-50 hour, filter, the clear paste that 55-70 ℃ of relative density of filtrate decompression simmer down to is 1.00-1.20, last activated-charcoal column is used earlier the distilled water eluting, colourless to eluent, reuse 9-11 doubly measures the 65-80% ethanol elution, collects eluent, and decompression recycling ethanol also concentrates, drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with an amount of water for injection; Other gets the Radix Notoginseng total arasaponins of recipe quantity, add an amount of water for injection dissolving, filter, Radix Notoginseng medicinal liquid and Fructus Gardeniae medicinal liquid mixing, regulate pH value to 5.5-7.5,0-4 ℃ cold preservation 20-30 hour, 0.45 μ m filter membrane filters, filtrate adds to the full amount of water for injection, filter with 0.22 μ m filter membrane, the filtrate fill becomes 10ml ampoule injection, 110-120 ℃ pressure sterilizing 20-40 minute, promptly.
The method of quality control that this compositions is made injection comprises discriminating and/or assay and/or finger printing.
Discrimination method comprises a kind of in the following method and/or two kinds:
A. get this product 5ml, water bath method, residue add dissolve with ethanol and quantitative the commentaries on classics is dissolved in volumetric flask 5ml, and other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 4mg, in contrast product solution; Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2000 VIB); Draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 8-11: 6-9: 1-3: 0.3-0.6 ethyl acetate-acetone-formic acid-water is developing solvent, launches, and takes out, and dries, and spray is with 5% vanillin sulfuric acid solution, heating; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; B. get this product 10ml, in addition water saturated n-butyl alcohol jolting is extracted 2-4 time, and each 10ml merges n-butanol extracting liquid, with the saturated water liquid washing of n-butyl alcohol 2-4 time, 100ml at every turn; Get the n-butanol layer reclaim under reduced pressure to doing, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets ginsenoside Rb
1, the ginsenoside Rg
1And Panax Notoginseng saponin R
1Reference substance adds methanol and makes the mixed liquor that every ml contains 6mg, in contrast product solution; Test according to thin layer chromatography (appendix IIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, below ambient temperature 15-25 ℃, with 11-15: 6-9: lower floor's solution that 1-3 chloroform-methanol-water is placed below 10 ℃ is developing solvent, launches, take out, dry, spray is with 1 → 10 sulphuric acid ethanol liquid, and it is clear to be heated to speckle colour developing in 105-115 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Content assaying method comprises a kind of in the following method and/or two kinds: a. jasminoidin, measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 D); Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 13-17: the 80-90 acetonitrile-water is a mobile phase; Detecting wavelength is that the 238nm theoretical cam curve should be not less than 1500 by the calculating of jasminoidin peak; It is an amount of that the preparation of reference substance solution, precision take by weighing the jasminoidin reference substance, adds methanol and make dissolving in right amount, makes the solution that every 1ml contains 0.14-0.15mg, promptly; The preparation of need testing solution, accurate this product 1ml that draws, water bath method, residue adds anhydrous alcohol solution, and ethanol liquid is transferred to another crucible, water bath method, residue is with dissolve with methanol and be transferred in the 100ml measuring bottle, adds methanol to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject high performance liquid chromatograph, measure, promptly; This product contains jasminoidin (C
17H
24O
10) every ml must not be less than 4.6-4.9mg; B. arasaponin is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 D); Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With acetonitrile-water binary gradient is mobile phase (seeing Table 1); The detection wavelength is 203nm; Theoretical cam curve is with the ginsenoside Rg
1The peak calculates should be not less than 2500; The reference substance solution preparation, precision takes by weighing arasaponin R1, ginsenoside Rg
1, ginsenoside Rb
1Add dissolve with methanol in right amount, make every 1ml and contain Panax Notoginseng saponin R
11.5mg, the ginsenoside Rg
12.8mg, ginsenoside Rb
14.1mg the mixing reference substance solution; The need testing solution preparation, the accurate sample 5ml that draws extracts three times with the water-saturated n-butanol jolting, each 10ml; N-butanol layer merges, and with the washing of 10ml n-butyl alcohol saturation water once, abandons water layer, the n-butanol layer reclaim under reduced pressure, and to doing, residue, shakes up to 5ml with dissolve with methanol, filters with microporous filter membrane 0.45 μ m, promptly; Algoscopy, the accurate reference substance solution 3 μ l that draw, 7 μ l, need testing solution 10 μ l inject high performance liquid chromatograph, calculate with the external standard two-point method, promptly; The every ml of this product contains Panax Notoginseng saponin R
1Must not be less than 0.18-1.25mg; The ginsenoside Rg
1Must not be less than 0.75-0.85mg; Ginsenoside Rb
1Must not be less than 1.8-2.5mg;
Table 1: acetonitrile-water binary gradient elution table time (min) flow velocity (ml/min) A (water) B (acetonitrile) 0 1.0 80 2,020 1.0 60 40
Injection method of quality control of the present invention also can adopt the method for finger printing, this method comprise a kind of in the following method and or two kinds:
A. the preparation of need testing solution: the accurate injection 5ml that draws, extract twice with water saturated n-butyl alcohol jolting, for the first time 10ml; 5ml merges n-butanol extracting liquid for the second time; With the saturated water 10ml backwash of n-butyl alcohol once, discard water layer, the n-butanol extracting liquid reclaim under reduced pressure is to doing, and residue is with dissolve with methanol and be settled in the 5ml volumetric flask, shakes up, and the 0.45um filter membrane filters, promptly; The preparation of reference substance solution: get the jasminoidin reference substance, the solution of making 0.5mg/ml with methanol is product solution in contrast; Assay method: detect wavelength 238nm, the acetonitrile-water gradient elution sees Table 2; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 101, writes down 60 minutes chromatogram, promptly; With the area at the retention time of the chromatographic peak (S peak) of jasminoidin and peak is 1 to calculate relative retention time and peak area ratio; Finger printing and technical parameter; Be 60 minutes writing time; The demarcation (relative retention time) of total fingerprint peaks: it is as follows to demarcate total fingerprint peaks according to the testing result of 10 batches of test samples: 1 (0.404) 2 (0.462) 3 (0.510) 4 (0.589) 5 (0.618) 6 (0.666) 7 (0.722) 8 (0.798) S (1.000) 9 (1.085) 10 (1.110) 11 (1.197) 12 (1.238) 13 (1.277); The ratio of total fingerprint peaks area: 1: S=1: (0.060-0.113); The part peak shape is described: separate when bad, peak 4 is the acromion at peak 5; Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak gross area must not be greater than 5% of peak area;
Instrument reagent high performance liquid chromatograph: Waters2695 pump; Waters 2996 UV-detector; The Millcnnium32 chromatographic work station; Diamonsil C18 post (5 μ m, 4.6mm * 150mm); Acetonitrile is chromatographically pure (U.S. J.T.Baker); The WAHAHA pure water; Methanol is chromatographically pure (Beijing prosperousization Fine Chemical Works);
Table 2 eluent gradient eluting table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
B. the preparation of need testing solution: the accurate injection 10ml that draws, divide the secondary jolting to extract with water saturated n-butyl alcohol 20ml, merge n-butanol extracting liquid; Saturated water 300ml divides three backwashes with n-butyl alcohol, discards water layer, and the n-butanol layer reclaim under reduced pressure is to doing, and residue is with dissolve with methanol and be settled in the 2ml measuring bottle, shakes up, and the 0.45um filter membrane filters, promptly; Preparation according to thing: get the ginsenoside Rg
1Reference substance (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute), the solution of making 2mg/ml with methanol is product solution in contrast; Assay method: detect wavelength 210nm, sensitivity 0.1AUFS: acetonitrile-water gradient elution, (seeing Table 3); Theoretical cam curve is by the ginsenoside Rg
1Calculate, should not hang down 30000; The accurate need testing solution 25 μ l that draw inject high performance liquid chromatograph, write down 70 minutes chromatogram, promptly; With the ginsenoside Rg
1The retention time of chromatographic peak (S peak) and the area at peak be 1 to calculate relative retention time and peak area ratio; Finger printing and technical parameter; Be 70 minutes writing time; The demarcation (relative retention time) of total fingerprint peaks: it is as follows to demarcate total fingerprint peaks according to the testing result of 10 batches of test samples: 1 (0.877) S (1.000) 2 (1.027) 3 (1.555) 4 (1.592) 5 (1.652) 6 (1.675) 7 (1.708) 8 (1.744) 9 (1.799) 10 (2.005) 11 (2.042) 12 (2.074) 13 (2.111) 14 (2.150) 15 (2.212) 16 (2.235) 17 (2.364) 18 (2.391) 19 (2.420) 20 (2.440) 21 (2.771); The ratio of total fingerprint peaks area: S: 3: 8: 21=1: (0.222-0.412): (0.405-0.735): (1.428-2.380); Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak gross area must not be greater than 5% of the gross area;
Instrument reagent: Waters 2695 Waters 2996 UV-detector Millcnnium32 chromatographic work station Symmetryshild
TMRP
18(5 μ m, 4.6mm * 250mm); Other is analytical pure for Fisher acetonitrile (U.S.'s chromatographically pure) Fisher methanol (U.S.'s chromatographically pure) WAHAHA pure water
Table 3 eluent gradient eluting table
Time (min) flow velocity (ml/min) A-water B-acetonitrile
0 1.0 83 17
20 1.0 76 24
40 1.0 60 40
50 1.0 50 50
55 1.0 0 100
60 1.0 0 100
70 1.0 0 100
The clinical injection that cures mainly the acute ischemic apoplexy of present composition preparation (collateral dredging is rescued injection of brain liquid), effect with removing pathogenic heat from blood and toxic substance from the body, disperse blood stasis and dredge collateral, by preventing the ischemia cascade reaction of apoplexy, improve brain slight irrigation stream, reach the therapeutic purposes of anti-neuron infringement;
Rescue the relevant pharmacodynamics test research of injection of brain lyolysis poison effect with collateral dredging, adopt ferric chloride to cause the thrombotic cerebral ischemia model of intraluminal middle cerebral artery occlusion in rats, studied collateral dredging and rescued rat nervous symptoms, brain water content, infraction cerebral morphology, cerebral tissue SOD and MDA content, cerebral infarction kitchen range Glu and NMDA expression, the Delayed Rectifier Potassium Current (I of injection of brain liquid this model
N) the mechanism of action; Experimental result demonstration collateral dredging is rescued the content that injection of brain liquid can obviously reduce the MDA of MCAT rat cerebral tissue, and increased SOD content reduces the expression of infraction cerebral tissue Glu and NMDA, increases Delayed Rectifier Potassium Current (I
N), reduce rat operation side brain water content, improve nervous symptoms;
Rescue the relevant pharmacodynamics test research of injection of brain liquid collateral dredging effect with collateral dredging, adopt ferric chloride to cause the thrombotic cerebral ischemia model of intraluminal middle cerebral artery occlusion in rats, studied collateral dredging and rescued of the effect of injection of brain liquid this rat model cerebral blood flow, anesthetized dog cerebral blood flow, blood stasis hemorheology of rat, chick chorioallantoic membrane angiogenesis; The result shows that collateral dredging rescues injection of brain liquid and can promote the generation of neovascularity, obviously reduce Blood stasis rat whole blood viscosity and plasma viscosity, and red cell deformability is strengthened, and aggregation descends, and improves anesthetized dog, MCAT rat brain blood flow, thereby realizes its collateral dredging function;
Drug action and XUESHUANTONG that collateral dredging is rescued injection of brain liquid compare, in the determination experiment to MCAT rat cerebral tissue water content, collateral dredging is rescued injection of brain liquid small dose group (comparing P＜0.01 with model group) obviously because XUESHUANTONG group (comparing not statistically significant with model group); Collateral dredging rescues that dosage group SOD obviously raises in the injection of brain liquid, with XUESHUANTONG group P＜0.05 relatively); Collateral dredging is rescued injection of brain liquid small dose group erythrodegeneration significantly increases (comparing P＜0.05, P＜0.01 with model group), and XUESHUANTONG does not have obvious effect to red blood cell deformation; Collateral dredging is rescued the heavy dose of group of injection of brain liquid blood vessel hyperplasia and significantly (is compared P＜0.05, P＜0.01 with model group), and a little less than the XUESHUANTONG effect; XUESHUANTONG can increase Delayed Rectifier Potassium Current (I
N), and that collateral dredging is rescued the effect of injection of brain liquid is not obvious, studies confirm that Delayed Rectifier Potassium Current (I at present
N) enhancing can promote the neuronal apoptosis that ischemia causes; The effect that collateral dredging is rescued injection of brain liquid in other effect experiment all is better than XUESHUANTONG; Collateral dredging is rescued injection of brain liquid and is obviously had the effect that promotes the anesthetized dog cerebral blood flow; In a word, collateral dredging rescues injection of brain liquid and XUESHUANTONG compares, and the reduction dosage is arranged, thereby reach the purpose of attenuation synergistic;
The experimental result summary is as follows:
1. collateral dredging is rescued the influence of injection of brain liquid to MCAT rat nervous symptoms
Experiment has adopted ferric chloride to cause the thrombotic rat cerebral ischemia model of middle cerebral artery, MCAT rat nervous symptoms evaluation results shows after the modeling: the rat that collateral dredging is rescued injection of brain liquid 83.2,41.6,20.8mg/kg group (20,10,5 times of quite clinical people's consumption respectively) is 12h after surgery, 24h, its nervous symptoms of 48h all has improve (P＜0.01, P＜0.05) in various degree; 2. collateral dredging is rescued the influence of injection of brain liquid to MCAT rat cerebral tissue water content
The rat operation side brain water content that 48h after the modeling, collateral dredging rescue injection of brain liquid 83.2,41.6,20.8mg/kg group is starkly lower than model group, and the difference of comparing with model group has significance (P＜0.01, p＜0.05);
3. collateral dredging is rescued injection of brain liquid to the morphologic influence of MCAT rat cerebral tissue
Cerebral tissue pathomorphology HE, the Nissl demonstration of dyeing, model group rat brain cortex ischemic focus cell quantity obviously reduces, the cell space atrophy, degeneration, painted shallow; Collateral dredging is rescued three dosage groups of injection of brain liquid rat cerebral ischemia kitchen range cell quantity than the model group showed increased, and the cellular atrophy degeneration obviously alleviates than model group; Graphical analysis shows that collateral dredging is rescued injection of brain liquid group positive cell surface density and model group relatively has statistical significance (P＜0.01), and the prompting collateral dredging is rescued injection of brain liquid and had and alleviate the effect that cerebral ischemia institute neurocyte extremely damages;
4. collateral dredging is rescued the influence of injection of brain liquid to SOD of MCAT rat cerebral tissue and MDA
Collateral dredging is rescued injection of brain liquid 83.2,41.6mg/kg organizes the content that can obviously reduce the MDA of MCAT rat cerebral tissue, increased SOD content (P＜0.01); The prompting collateral dredging is rescued injection of brain liquid and is had the free radical resisting damage, the effect of protection cerebral tissue;
5. collateral dredging is rescued the influence of injection of brain liquid to MCAT rat infraction cerebral tissue Glu and NMDA expression
Adopt ferric chloride to cause thrombotic rat cerebral ischemia model of middle cerebral artery and immunohistochemical method, the result shows that collateral dredging rescues injection of brain liquid and can obviously reduce Glu, the NMDA expression at focus of infarct;
6. collateral dredging is rescued the influence of injection of brain liquid to Delayed Rectifier Potassium Current
Use the patch-clamp electrophysiological technique, detected collateral dredging and rescued the influence of injection of brain liquid to the myocardial cell Delayed Rectifier Potassium Current, the result shows that collateral dredging is rescued injection of brain liquid to Delayed Rectifier Potassium Current (I
N) influence not obvious;
7. collateral dredging is rescued the influence of injection of brain liquid to MCAT rat brain blood flow
MCAT rat cerebral tissue blood flow obviously reduces, and collateral dredging is rescued the cerebral tissue blood flow of injection of brain liquid 83.2,41.6mg/kg group modeling 48h all apparently higher than model group (P＜0.01), illustrates that this medicine is to the effect of having clear improvement of ischemia hindbrain tissue blood flow; Collateral dredging is rescued injection of brain liquid the rat Blood stasis that acute stress causes is improved significantly, and low dose of effect is the most obvious;
8. collateral dredging is rescued the influence of injection of brain liquid to the anesthetized dog cerebral blood flow
Experiment is carried out 33 healthy hybrid dogs, and test shows: collateral dredging is rescued injection of brain liquid can significantly improve anesthetized dog cerebral blood flow (P＜0.05 or P＜0.01); It acts on beginning onset in 10 minutes behind the injectable drug, and drug effect was maintained to after the administration 50 minutes; Collateral dredging is rescued injection of brain liquid 83.2, the 41.6mg/kg group does not relatively have significant difference with the positive drug XUESHUANTONG; Collateral dredging is rescued injection of brain liquid low dose group and positive drug XUESHUANTONG relatively, and significant difference (P＜0.05) was arranged behind injectable drug in 45 minutes and 50 minutes;
9. collateral dredging is rescued the influence of injection of brain liquid to the stasis syndrome hemorheology of rat
Collateral dredging is rescued dosage in the injection of brain liquid (41.6mg/kg) can obviously reduce Blood stasis rat whole blood viscosity (with model group comparison P＜0.01 with low dose of (20.8mg/kg), P＜0.001, P＜0.0001), plasma viscosity (but not seeing significant difference) on a declining curve, small dose group obviously reduce packed cell volume (comparing P＜0.0001 with model group); Small dose group can make red cell deformability strengthen, aggregation decline (P＜0.01, P＜0.001);
10. collateral dredging is rescued the influence of injection of brain liquid to the chick chorioallantoic membrane angiogenesis
Collateral dredging is rescued injection of brain liquid 83.2,41.6, the 20.8mg/kg group all has the effect that promotes the chick chorioallantoic membrane angiogenesis;
Below to above-mentioned part experiment describe in detail:
Experimental example one collateral dredging is rescued the influence of injection of brain liquid to MCAT rat nervous symptoms
Experiment material
1. medicine and reagent
Be subjected to reagent: collateral dredging is rescued injection of brain liquid and provided by Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia, color: outward appearance is faint yellow, content: 10.39mg/ml, have removing pathogenic heat from blood and toxic substance from the body, and the function of disperse blood stasis and dredge collateral cures mainly the acute ischemic apoplexy; Lot number: 2001042322; Clinical plan consumption: 4.16mg/kg, intravenous injection;
Positive control drug: XUESHUANTONG ZHUSHEYE is provided by Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia, color: white, and content: 50mg/ml has the function of blood circulation promoting and blood stasis dispelling, cures mainly the acute ischemic apoplexy; Lot number: in defend the accurate word (1996) of medicine No. 001945; Clinical plan consumption: 10mg/kg, intravenous injection 1～2 time/day, 1 5ml;
Reagent and medicine: FeCl
36H
2O (A.R.), Beijing Chemical Plant's product is with the preparation of 1mol/L hydrochloric acid;
2. animal
The Wistar rat, the male and female dual-purpose, body weight 180～200g is provided by Chinese Academy of Medical Sciences's animal center breeding field, the quality certification number: the moving word of doctor 01-3008 number;
3. instrument
The XTT anatomic microscope, Beijing electric light scientific instrument factory product; SHZ-22 type water bath with thermostatic control agitator, granary, Jiangsu medical apparatus and instruments factory product; AEG-220 type electronic analytical balance, day island proper Tianjin instrument company product;
Method and result
1. grouping and administration
60 rats are divided into six groups at random, be that sham operated rats, MCAT model group, collateral dredging are rescued injection of brain liquid 83.2mg/kg group, collateral dredging and rescued injection of brain liquid 41.6mg/kg group, collateral dredging and rescue injection of brain liquid 20.8mg/kg group (be equivalent to respectively clinical people's consumption 20,10,5 times), XUESHUANTONG 50mg/Kg group (quite 5 times of people's consumption), 10 every group; Tail intravenously administrable after the modeling, a daily dose gives at twice, is administered five times altogether;
2. modeling method
The anesthesia of rats by intraperitoneal injection 10% chloral hydrate solution (350mg/kg); Press Tamura
[4]Deng method, improve a little; The rat right arm reclining is fixed, make a curved incision at paropia and external auditory meatus line mid point, be about 1.5cm, pinch off temporalis and excision, expose temporal bone, make a diameter 2.5mm bone window at cheekbone and temporo squamosum joint near oral-lateral 1mm place with dental burr, the cleaning residue exposes middle cerebral artery (between tractus olfactorius and inferior cerebral vein); Put small pieces hollow plastic thin film protection blood vessel surrounding tissue; There is the small pieces quantitative filter paper of 50% ferric chloride solution, 10 μ l to apply on this section middle cerebral artery suction
(6), take off filter paper behind the 30min, use the normal saline flushing local organization, layer-by-layer suture steams again and raises; Sham operated rats is except that not dripping the ferric chloride solution the same model group of all the other operating procedures;
3.MCAT the evaluation of rat nervous symptoms
(12h, 24h 48h), press Bederson etc. to different time after surgery
(7)Method and improved, animal is carried out behavior scoring; 1. carry the about chi of Mus tail built on stilts, observe forelimb flexing situation; Stretch to ground as two forelimb symmetries, be designated as 0 fen; As the flexing that shoulder flexing, elbow flexing, shoulder inward turning or existing wrist elbow appear in the offside forelimb of performing the operation has inward turning person again, is designated as 1,2,3 and 4 fen; 2. animal is placed on the level and smooth ground, push away both shoulders respectively, check resistance to side shifting; Be designated as 0 fen as bilateral resistance equity and strong person; As resistance descender when the operation offside promotes, according to decline degree difference be divided into gently, in, weigh 3 and spend, be designated as 1,2,3 fen respectively; 3. animal two forelimbs are put on the wire netting, observed the muscular tension of two forelimbs; Bilateral muscular tension equity and strong person are 0 minute; Be designated as 1,2,3 fen according to operation offside muscle of anterior limb tension force decline degree difference equally; 4. carry the about chi of Mus tail built on stilts, animal has ceaselessly to operation offside revolver, is designated as 1 fen; According to above standard scoring, full marks are 11 minutes, and mark is high more, and the behavior disorder of animal is serious more; To behavior detect the marking value organize between relatively, the t check; The results are shown in Table 1;
Table 1 collateral dredging is rescued injection of brain liquid to the influence of MCAT rat nervous symptoms (X ± SD)
Mental symptom scoring group dosage mg/kg N
12h 24h 48h sham operated rats-10 00 0MCAT model group-10 5.5 ± 0.55 4.67 ± 0.52 4 ± 0.63 collateral dredging is rescued brain group 83.2 10 4.33 ± 0.82**, 3.83 ± 0.41** 3.17 ± 0.75*
41.6 10 4.4±0.55** 3.83±0.75** 3.33±0.82*
20.8 10 4.67 ± 0.55 4 ± 0.63*, 3.33 ± 1.03 XUESHUANTONG groups, 50 10 3.67 ± 0.52**, 4 ± 0.63*, 3.33 ± 0.52*
Annotate: each group is compared * P＜0.05, * * P＜0.01 with model group;
The result shows, removes sham operated rats and does not see that dystropy changes, and MCAT model group rat is 12h after surgery, and hemiplegia sample symptom all appears in 24h, 48h, mainly show as in the operation offside forelimb to receive, and the shoulder inward turning, muscle of anterior limb tension force reduces, and the shoulder drag descends; Collateral dredging rescue injection of brain liquid 20.8mg/kg organize after surgery the 48h nervous symptoms improve not obvious outside, the rat that collateral dredging is rescued each dosage group of injection of brain liquid and XUESHUANTONG group is 12h after surgery, 24h, its nervous symptoms of 48h all have improve (P＜0.01, P＜0.05) in various degree;
Experimental example two collateral dredging are rescued the influence experiment material of injection of brain liquid to MCAT rat cerebral tissue water content
1. medicine and reagent
Be subjected to reagent, positive control drug with test 20.1;
2. animal
The Wistar rat, the male and female dual-purpose, body weight 180～200g, is provided the quality certification number by 71 by Chinese Academy of Medical Sciences's animal center breeding field: the moving word of doctor 01-3008 number;
3. instrument
The XTT anatomic microscope, Beijing electric light scientific instrument factory product; AEG-220 type electronic analytical balance, day island proper Tianjin instrument company product; Df-206 type air dry oven, west city, Beijing medical apparatus and instruments factory product; Method and result
Grouping, administration and operation are with test 20.1; 48 hours broken ends of postoperative are got brain, cut brain mid portion (front and back are respectively removed 3 millimeters), divide right and left, and blot surface moisture with filter paper, respectively weighing left and right sides brain sheet weight in wet base; Place baking oven again, 105 ℃ are toasted 48 hours to constant weight, and accurately the weighing dry weight is calculated water content
[8], with control sides comparison on the same group, and compare between group, carry out the t check; The results are shown in Table 2;
Table 2 collateral dredging is rescued injection of brain liquid to the influence of MCAT rat cerebral tissue water content (X ± SD)
Dosage water content (%)
Group N
(mg/kg) the right brain water content of left brain water content
Sham operated rats-10 77.30 ± 0.79 77.09 ± 0.57
△ △
MCAT model group-10 77.04 ± 0.48** 78.90 ± 0.96
Collateral dredging is rescued brain group 83.2 10 76.55 ± 1.33** 78.09 ± 1.03
41.6 10 76.77±0.78** 77.96±0.91
20.8 10 77.51±2.16** 75.11±2.03
△△
XUESHUANTONG group 50 10 77.04 ± 0.48** 78.90 ± 0.96
Annotate: compare with model group
△ △P＜0.01; Compare * * P＜0.01 with offside brain water content on the same group;
The result shows, postoperative 48h, sham operated rats is not seen the big brain water content abnormal change in the left and right sides, model group rat operation side brain water content obviously increases, the rat operation side brain water content that collateral dredging is rescued each dosage group of injection of brain liquid, XUESHUANTONG group obviously is less than model group, compare with model group and to have significant difference (P＜0.01), but each administration group operation side brain water content is compared with left brain and obviously increased (P＜0.01); Experimental example 3 collateral dredging are rescued the influence experiment material of injection of brain liquid to SOD of MCAT rat cerebral tissue and MDA
1, medicine and reagent
Be subjected to reagent: XUESHUANTONG ZHUSHEYE, collateral dredging are rescued injection of brain liquid with experiment 20.1;
MDA, SOD reagent close available from Nanjing and build up bio-engineering research institute; Lot number: 20011031;
2, animal
The Wistar rat, male, body weight 190～210g, 60, for Chinese Academy of Medical Sciences's animal center breeding field provides, the quality certification number: the moving word of doctor 01-3008 number;
3, instrument
721 type spectrophotometers, Shanghai the 3rd analytical tool factory product; Method and result
Grouping, administration, modeling method are with experiment 20.1; Modeling, 24 hours broken ends of administration are got brain, and decerebellation, olfactory bulb and brain stem add 9 times of normal saline and be prepared into 10% brain homogenate, and be standby;
1, the mensuration of SOD
Close explanation by SOD reagent and measure the SOD vigor, the results are shown in Table 3;
2, the mensuration of MDA
Close explanation by MDA reagent and measure MDA content, the results are shown in Table 3;
Table 3 collateral dredging is rescued injection of brain liquid to the influence of SOD of MCAT rat cerebral tissue and MDA (X ± SD)
Group dosage mg/kg N MDAmmol/g cerebral tissue SODnu/mg cerebral tissue
Sham operated rats-10 0.54 ± 0.11**, 144 ± 20.20**
Model group-10 0.87 ± 0.21 92.17 ± 21.54
83.2 10 0.56±0.1** 127.67±20.55**
Collateral dredging is rescued brain group 41.6 10 0.59 ± 0.12**, 138.33 ± 27.27**
20.8 10 0.79±0.21 98.5±29.04
XUESHUANTONG group 50 10 0.57 ± 0.07** 114.5 ± 21.86**
Annotate: each group is compared * * P＜0.01 with model group;
The result shows that rat is after closing middle cerebral artery with fixed attention, and its superoxide dismutase (SOD), MDA all have significant change, model group rat MDA content is apparently higher than sham operated rats, SOD is starkly lower than sham operated rats, and collateral dredging is rescued injection of brain liquid can obviously reduce MDA content, increased SOD vigor (P＜0.01); Point out this medical instrument that the effect of free radical resisting injury protection cerebral tissue is arranged;
Experimental example four-way network is rescued the influence experiment material of injection of brain liquid to rats with cerebral ischemia brain cortex focus of infarct Glu and NMD expression
1, medicine and reagent
Be subjected to reagent: collateral dredging is rescued injection of brain liquid and is provided by Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia, cures mainly the acute ischemic apoplexy;
Positive control drug: XUESHUANTONG ZHUSHEYE is provided by Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia, cures mainly the acute ischemic apoplexy, intravenous injection, 2 times/day, 1 5ml; Lot number: 2000091112;
Reagent and medicine: FeCl
36H
2O (A.R.), Beijing Chemical Plant's product is with the preparation of 1mol/L hydrochloric acid;
Immunohistochemistry reagent: the anti-Glu of rabbit, NMDA (1: 200, Santa cruz company), biotinylation goat anti-rabbit igg and ABC complex (1: 200, Histostatn-sp test kit, ZYMED company);
2, animal
The Wistar rat, the male and female dual-purpose, body weight 180～200g is provided by Chinese Academy of Medical Sciences's animal center breeding field, the quality certification number: the moving word of doctor 01-3008 number;
3, instrument
The POLYVAR all purpose microscope, U.S.'s product; C
8Very color pathological image analyser, BJ University of Aeronautics ﹠ Astronautics's product; Test method
1. grouping and administration
60 rats are divided into six groups at random, be that sham operated rats, MCAT model group, collateral dredging are rescued injection of brain liquid 83.2mg/kg group, collateral dredging and rescued injection of brain liquid 41.6mg/kg group, collateral dredging and rescue injection of brain liquid 20.8mg/kg group (be equivalent to respectively clinical people's consumption 20,10,5 times), XUESHUANTONG 50mg/kg group (quite 5 times of people's consumption), 10 every group; Tail intravenously administrable after the modeling, a daily dose gives at twice, is administered five times altogether;
2. modeling method
The anesthesia of rats by intraperitoneal injection 10% chloral hydrate solution (350mg/kg); Press Tamura
[3]Deng method, improve a little; The rat right arm reclining is fixed, make a curved incision at paropia and external auditory meatus line mid point, be about 1.5cm, pinch off temporalis and excision, expose temporal bone, make a diameter 2.5mm bone window at cheekbone and temporo squamosum joint near oral-lateral 1mm place with dental burr, the cleaning residue exposes middle cerebral artery (between tractus olfactorius and inferior cerebral vein); Put small pieces hollow plastic thin film protection blood vessel surrounding tissue; There is the small pieces quantitative filter paper of 50% ferric chloride solution, 10 μ l to apply on this section middle cerebral artery suction
(4), take off filter paper behind the 30min, use the normal saline flushing local organization, layer-by-layer suture steams again and raises; Sham operated rats is except that not dripping the ferric chloride solution the same model group of all the other operating procedures;
3. animal is handled
After the last administration one hour (being postoperative 48 hours), broken end is got brain, fix with 10% neutral formalin respectively, and paraffin embedding, immunohistochemical staining is made in section (getting optic chiasma front and back brain sheet), and the microscopic observation tissue morphology changes, and does graphical analysis;
4. the immunohistochemical staining method adopts the ABC method, and step is as follows: 1.. and 1% methanol H is gone in section
2O
2Tuck in, room temperature 30min is 2.. protease K digesting, 37 ℃ of 30min are 3.. normal sheep serum, room temperature 30min is 4.. the anti-Glu of rabbit, NMDA (1: 200, Santa cruz company), 5. 4 ℃ spent the night. biotinylation goat anti-rabbit igg and ABC complex (1: 200, Histostatn-sp test kit, ZYMED company), 6. .0.05%DAB-0.1%H of 37 ℃ of 2h
2O
2The liquid colour developing; 1., 2., 3., 5., 6. all wash 3 times before the step, wash 5min with 0.01MPBS at every turn; Negative control saves one and resists or resist with normal sheep serum replacement one;
5. graphical analysis and statistical procedures
To Glu, NMDA SABC result, analyze with the very color pathological image analytical system of CMIAS8 (Chinese Aero-Space university graphical analysis center), add up the surface density of each; Statistics are checked with t between group; Experimental result
1. collateral dredging is rescued the influence that injection of brain liquid is expressed rats with cerebral ischemia brain cortex focus of infarct Glu
Normal rat brain cortex Glu is expressed in the kytoplasm of cell, and the nuclear district is negative, and cellular morphology is cone-shaped more, and has projection, and cell is methodically arranged; The Glu cell quantity is than normal rat showed increased in the model group rat brain cortex focus of infarct, and Glu expresses also and obviously strengthens, and cellular morphology is the cone-shaped of atrophy degeneration; Glu expression ratio model group obviously weakens in positive drug group, heavy dose of group, the middle dosage group rat brain, and wherein middle dosage group Glu expresses the most weak, and cell quantity is minimum; The same model group of the expression of small dose group Glu; Graphical analysis statistical result sees Table 4;
Table 4 collateral dredging is rescued the influence that injection of brain liquid expresses MCAT rat cerebral cortex focus of infarct Glu (X ± SD)
Dosage
Group N positive cell surface density
(mg/kg)
Sham operated rats-9 0.09 ± 0.02**
MCAT model group-9 0.13 ± 0.02
Collateral dredging is rescued injection of brain liquid group 83.2 9 0.09 ± 0.02**
41.6 9 0.07±0.02**
20.8 9 0.06±0.02**
XUESHUANTONG group 50 9 0.10 ± 0.10**
Annotate: compare * * P＜0.01 with model group
2. collateral dredging is rescued the influence that injection of brain liquid is expressed rats with cerebral ischemia brain cortex focus of infarct NMDA
Normal rat brain cortex NMDA is expressed in the kytoplasm of cell, and the nuclear district is negative, and cellular morphology is cone-shaped and circle more, and cell is methodically arranged; The expression of the interior NMDA of rat model brain cortex focus of infarct is obviously strong than normal rat, and cellular morphology is swelling and shrinkage deformation form; Positive drug group and heavy dose of group, middle dosage group, small dose group rat NMDA expression ratio model group weaken, and the expression of wherein heavy dose of group, middle dosage group NMDA is the most weak, and heavy dose of group NMDA cell is cone-shaped, and the atrophy degeneration; Middle dosage group NMDA positive cell is small circular more; Statistical result sees Table 5;
Table 5 collateral dredging is rescued the influence that injection of brain liquid expresses MCAT rat cerebral cortex NMDA (X ± SD)
Group dosage (mg/kg) N positive cell surface density
Sham operated rats-9 0.08 ± 0.02*
MCAT model-9 0.11 ± 0.02
Collateral dredging is rescued injection of brain liquid group 83.2 9 0.09 ± 0.02**
41.6 9 0.07±0.02**
20.8 9 0.05±0.01**
XUESHUANTONG group 50 9 0.07 ± 0.01*
Annotate: compare * P＜0.05 with model group, material P＜0.01
Conclusion: collateral dredging is rescued injection of brain liquid by reducing Glu, the NMDA expression at focus of infarct, reverses swelling, the degeneration of cell, thereby reaches therapeutic purposes; Experimental example five-way network is rescued the influence experiment material of injection of brain liquid to the anesthetized dog cerebral blood flow
1. animal
33 of healthy hybrid dogs, the male and female dual-purpose, body weight 12～15kg, tonneau laboratory animal plant provides by the Haidian District, Beijing City, animal quality certification numbering: 024-069 number;
2. medicine
Collateral dredging is rescued injection of brain liquid: Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia (Gan Qi card pharmaceutical factory, the Inner Mongol) produces; Lot number: 2001042322; Specification: 20ml/ only; 263mg ginsenoside/20ml;
Blank medicine: normal saline;
XUESHUANTONG ZHUSHEYE (positive drug): Kangyuan Pharmaceutical Industrial Co Ltd, Inner Mongolia (Gan Qi card pharmaceutical factory, the Inner Mongol) produces; Lot number: 2000091112; Specification: 250mg/5ml
Medicine stock solution places 4 ℃ of stored refrigerated;
3. medicine preparation
When each experiment begins, carry out the medicine preparation according to the weight of animals:
XUESHUANTONG ZHUSHEYE: 0.5ml stock solution/kg; (25mg/kg)
Collateral dredging is rescued injection of brain liquid low dose group: and 0.394ml stock solution/kg (the 5.18mg ginsenoside/kg);
Collateral dredging is rescued dosage group in the injection of brain liquid: and 0.788ml stock solution/kg (the 10.4mg ginsenoside/kg);
Collateral dredging is rescued injection of brain liquid high dose group: and 1.577ml stock solution/kg (the 20.7mg ginsenoside/kg);
Draw the stock solution of corresponding milliliter number, be mixed into the intravenous drip liquid that cumulative volume is 100ml, heating in water bath to 37 ℃ laggard row vein instillation with an amount of normal saline;
4. experimental apparatus and apparatus
TA-4000 polygraph (U.S. GouldInc company product), MF-27 electromagnetic flowmeter (Japanese photoelectricity company product), animal surgery apparatus one cover;
5. experimental procedure laboratory animal operation
The animal pneumoretroperitoneum of weighing is injected 5% pentobarbital sodium 30mg/kg, and anesthesia back position is fixed, and it is standby to separate the left side femoral vein; Cervical incision separates right common carotid artery, and intubate (anticoagulant in the heparin solution pipe) directly writes down Blood pressure of carotid artery; Separate left common carotid artery, separate the ligation external carotid artery, common carotid artery is embedded the electromagnetic flowmeter probe, and (φ=2mm), fix perpendicular to common carotid artery surveys the ICAF amount fully and uses; Dry for preventing probe, drip normal saline and keep moistening; Lay ECG electrode, record standard limbs II lead electrocardiogram; Respiration probes is placed nostril place's recording respiration frequency and respiratory depth;
6. laboratory animal administration
Stablized after the operation 30 minutes, and write down every index, as contrasting before the administration; During the experiment beginning, carry out the preparation of medicinal liquid according to preceding method; Dog is at the uniform velocity instiled through the left side femoral vein and studies medicine accordingly, and record instils the time started, and the control medicine at the uniform velocity dripped off in 20 minutes;
7. observation index
Measure immediately when instiling beginning (0 minute), every 5 minutes records once, write down 15～30 seconds at every turn, chart drive speed is 10mm/s during record, and record interval chart drive speed is 50mm/h, and the polygraph monitor is monitored synchronously; Observed and recorded drug effect to the beginning of instiling finished in back 60 minutes; The following physical signs of synchronous recording:
1) ICAF amount
2) arteriotony experimental result
The TA-4000 polygraph on the recorder chart of special use, by artificial counting, is converted to numeral with the figure of record with Experiment Data Records, is attached to each experimental record back with the form of form;
Experimental result is used the SAS statistical software, carries out the t check to have determined whether significant difference with regard to the every index between each group;
Collateral dredging rescue injection of brain liquid to the influence of anesthetized dog cerebral blood flow referring to table 6;
Table 6 collateral dredging is rescued the influence (ml/min) of injection of brain liquid to the anesthetized dog cerebral blood flow
Collateral dredging is rescued the nicergoline network and is rescued the nicergoline network and rescue brain in the time of after the administration
XUESHUANTONG (n=6) normal saline (n=7)
Between dosage (n=7) low dosage (n=6) in (branch) high dose (n=7)
0 110.7±2.2 109.1±2.6 106.6±6.1 110.8±3.4 111.9±2.0
5 117.5±6.6 116.7±5.0 112.0±7.3 116.7±5.2 111.8±2.9
10 124.2±8.8
** 121.7±7.3
** 119.4±9.4
* 120.8±8.6
* 110.3±5.9
15 126.7±10.8
** 128.3±16.7
** 122.7±11.3
** 122.2±7.1
** 110.3±4.5
20 128.3±13.7
** 131.4±16.0
** 125.4±12.7
** 123.8±8.6
** 111.3±6.6
25 130.8±16.6
** 130.7±12.1
** 127.6±11.7
** 124.0±7.9
** 110.9±6.6
30 115.3±53.5 131.4±8.9
** 126.9±10.9
** 125.5±7.4
** 110.6±5.6
35 132.2±14.5
** 130.4±8.1
** 122.9±15.5 123.8±6.0
** 111.3±8.3
40 129.2±10.2
** 126.9±6.9
** 126.1±8.5
** 120.2±2.6 113.3±8.7
45 130.0±8.4
** 126.1±8.7
** 126.7±6.9
** 115.8±4.9
△ 113.0±7.9
50 130.2±7.8
** 126.1±12.8
** 125.6±6.8
** 115.3±11.9
△ 110.9±7.9
55 122.5±13.6 119.3±13.7 122.3±10.2 112.8±7.5
** 110.3±6.1
60 119.2±17.4 118.4±14.6 106.4±39.7 108.7±11.9 110.9±7.2
*Compare P＜0.05 with matched group;
*Compare P＜0.01 with matched group;
△Compare P＜0.05 with XUESHUANTONG;
Experiment shows: collateral dredging is rescued injection of brain liquid can significantly improve anesthetized dog cerebral blood flow (P＜0.05 or P＜0.01); It acts on beginning onset in 10 minutes behind the injectable drug, and drug effect was maintained to after the administration 50 minutes; Collateral dredging is rescued injection of brain liquid high dose group and middle dosage group and positive drug XUESHUANTONG does not relatively have significant difference; Collateral dredging is rescued injection of brain liquid low dose group and positive drug XUESHUANTONG relatively, and significant difference (P＜0.05) was arranged behind injectable drug in 45 minutes and 50 minutes;
2. collateral dredging is rescued the influence of injection of brain liquid to anaesthetized dog blood pressure
Collateral dredging rescue injection of brain liquid to the influence of anesthetized dog systolic pressure referring to table 7;
Table 7 collateral dredging is rescued the influence (mmHg) of injection of brain liquid to the anesthetized dog systolic pressure
The time collateral dredging is rescued the nicergoline network and is rescued the nicergoline network and rescue the brain normal saline after the administration
XUESHUANTONG (n=6)
Dosage (n=7) low dosage (n=6) (n=7) in (branch) high dose (n=7)
0 148.3±12.5 149.6±12.5 140.6±16.8 146.8±19.6 137.2±17.0
5 146.3±13.6 152.1±11.8 138.0±11.7 147.9±18.9 138.0±18.4
10 149.8±16.7 153.0±13.3 139.0±11.7 149.3±18.1 137.4±17.3
15 148.7±15.2 154.7±13.5 140.0±10.8 149.3±18.8 136.9±16.6
20 147.7±16.5 152.7±12.6 140.6±10.7 149.3±19.5 137.1±15.9
25 148.2±16.0 151.0±13.1 139.1±10.9 149.5±18.3 136.7±18.3
30 144.3±11.2 151.9±12.4 139.1±10.4 148.8±18.6 137.9±.16.6
35 144.3±11.5 150.4±13.1 139.9±9.5 150.3±19.3 138.1±17.6
40 147.2±12.6 147.6±12.6 140.0±13.7 148.2±19.1 138.4±16.6
45 146.7±13.8 147.4±13.0 141.7±10.7 144.6±22.3 138.7±16.3
50 144.0±10.4 146.7±14.7 141.4±12.2 147.3±19.9 138.8±17.0
55 146.7±16.9 145.4±16.3 140.6±12.9 143.5±17.2 137.8±17.0
60 148.2±12.1 146.8±14.9 141.1±13.2 145.4±18.6 138.2±17.1
Experiment shows: collateral dredging is rescued injection of brain liquid does not have significant difference (P＞0.05) to the influence of anesthetized dog systolic pressure
Collateral dredging rescue injection of brain liquid to the influence of anesthetized dog diastolic pressure referring to table 8;
Table 8 collateral dredging is rescued the influence (mmHg) of injection of brain liquid to the anesthetized dog diastolic pressure
The time collateral dredging is rescued the nicergoline network and is rescued the nicergoline network and rescue the brain normal saline after the administration
XUESHUANTONG (n=6)
Dosage (n=7) low dosage (n=6) (n=7) in (branch) high dose (n=7)
0 116.3±10.2 121.3±10.5 112.9±14.2 119.7±13.2 109.6±13.1
5 115.0±8.6 119.7±11.6 111.2±10.4 120.0±12.9 110.8±14.7
10 119.5±14.4 120.0±10.9 112.1±10.6 121.3±13.5 110.3±13.5
15 116.0±7.9 119.4±13.9 112.4±10.0 120.5±12.7 110.3±13.5
20 116.8±10.6 118.1±12.3 112.8±10.6 119.7±13.1 109.9±13.1
25 117.7±9.8 117.0±12.8 112.0±10.7 120.8±12.4 110.7±14.1
30 116.0±8.0 117.6±13.0 112.1±10.6 119.7±13.0 110.3±14.0
35 115.7±8.0 118.4±12.6 113.0±9.4 122.8±11.9 110.4±14.7
40 116.7±8.4 115.6±12.8 112.1±12.1 121.8±12.8 110.9±13.2
45 117.2±9.2 117.0±12.2 113.7±10.5 117.8±16.7 111.1±12.2
50 114.8±8.5 116.0±14.9 113.0±12.1 118.7±17.0 111.1±13.5
55 118.7±13.3 115.5±16.0 112.6±13.5 117.7±15.5 110.7±13.7
60 117.3±10.1 117.0±13.9 112.9±13.4 116.3±15.8 110.7±13.7
Experiment shows: collateral dredging is rescued the high, medium and low dosage group of injection of brain liquid not to be had significant difference (P＞0.05) experimental example clematis stem network to rescue injection of brain liquid to the influence of anesthetized dog diastolic pressure the chick chorioallantoic membrane angiogenesis influenced experiment material and method
1. medicine:
Injection Chinese medicine XUESHUANTONG, collateral dredging are rescued brain and are provided by professor Niu Xin, and concentration is 50mg Radix Notoginseng total arasaponins/ml solution; This medicinal liquid is divided into greatly (50mg Radix Notoginseng total arasaponins/ml), in (25mg Radix Notoginseng total arasaponins/ml), little (three concentration of 12.5mg Radix Notoginseng total arasaponins/ml);
2. hatch:
Fresh Rhizoma Euonymus hatching egg is available from Changping livestock corporation, select no damaged crackle, the approximate hatching egg that air chamber is arranged of size, after the ethanol with 75% is sterilized its surperficial wiping, greatly head-up, tilt to put into 37.8 ℃ of incubators of disinfectant, place water pond in the incubator to keep certain humidity in the case; For preventing embryo's adhesion, promote the amniotic membrane motion, every day, turning egg(s) was 4-5 time
[3]
3. window:
With reference to paying method such as think of a way
[4]And improved; Take out when hatching egg is hatched to the 5th day, on candler, check the vascular development situation; Select clear, the well-developed hatching egg of blood vessel, between two trunks in the lower right of embryo head, draw the window's position of fixed one 1cm * 1cm, and also draw a labelling at the air chamber place; In the immigration super-clean bench, with finish position hatching egg shell with 75% ethanol wiping sterilization after, bore an aperture at hatching egg air chamber place to be used for decompression with dental burr earlier, the little reduction of reuse dental burr is cut the shell that picture is decided the window's position, remove the place's chorion of windowing gently with the ophthalmology tweezer, expose white shell membrane, carefully remove membrana putaminis behind a little normal saline again and expose chorioallantoic membrane;
4. dosing:
According to results of study such as Zhang Shucheng
[5], we adopt gelfoam as adding drug carrier; Sterile gelatin sponge (available from China-Japan Friendship Hospital) is cut into the fritter of 5mm * 2mm * 2mm, add 35 μ l with the filterable medicinal liquid of 0.22 μ m sterile filters after, place on the chorioallantoic membrane of window middle position, seal the aperture at window and air chamber place with aseptic adhesive tape after, put back to and continued in the incubator to hatch 48 hours; And matched group (physiological saline solution, Embryo Gallus domesticus of 35 μ l/), the positive group of XUESHUANTONG are set, collateral dredging is rescued injection of brain liquid 83.2mg/kg, 41.6mg/kg, 20.8mg/kg group; All do 20 Embryo Gallus domesticus for every group;
5. get film:
Open the adhesive tape on the Embryo Gallus domesticus fenestella, bore an aperture in the hatching egg bottom, wait to flow out the Ovum Gallus domesticus album about 1ml after, in window, drip fixative (methanol: acetone=1: 1), fixing 15-20min at room temperature; Afterwards, eggshell is broken into two with one's hands to both sides, content is all poured in the culture dish, carefully chorioallantoic membrane is taken off with eye scissors and ophthalmology tweezer, puts into the plate that fills clear water and launches, and apply ointment or plaster on filter paper the preservation of drying in the shade;
6. count:
Under anatomical lens, observe vessel growth situation and counting on the chorioallantoic membrane; Count range: with the administration be the center in the scope of diameter 1.5cm, be divided into large, medium and small blood vessel and count according to the diameter of blood vessel;
7. meter is learned and is handled:
We carry out statistical procedures with physiology saline group with the t check with count results; Experimental result
The data statistics result is as shown in table 9:
Table 9 collateral dredging is rescued injection of brain liquid to the angiopoietic influence of chick chorioallantoic membrane
Dosage chorioallantoic membrane blood vessel number/embryo (x ± s)
Group
The little blood vessel of mg/kg trunk medium vessels
Matched group 2.33 ± 1.53 11.00 ± 1.00 43.67 ± 6.35
Collateral dredging is rescued brain group 83.2 2.00 ± 0.82 8.86 ± 2.27 35.43 ± 6.60
41.6 1.71±0.76 7.86±2.12* 36.14±2.34**
20.8 2.29±1.11 9.43±2.30 36.71±3.86*
XUESHUANTONG group 50 2.01 ± 0.55 6.80 ± 3.11*, 30.60 ± 6.77**
Annotate: n=20, compare with matched group: * P＜0.05 * * P＜0.01
The result shows that collateral dredging rescues each dosage group of injection of brain liquid, and particularly heavy dose of group and XUESHUANTONG relatively have the effect that promotes angiogenesis;
Following embodiment all can realize the effect of above-mentioned experimental example.
Embodiment 1Injection
Fructus Gardeniae 175g Radix Notoginseng total arasaponins 5.5g
Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, the percolation under the photograph fluid extract item (" appendix IO of Chinese pharmacopoeia version in 2000), with 70% ethanol percolation of 8 times of amounts, collect percolate, reclaim ethanol, being concentrated into relative density is the clear paste of 1.10 (60 ℃ of mensuration), adds 4 times of water gaging dilutions, stir evenly, 0-4 ℃ of cold preservation 48 hours filters, and filtrate decompression simmer down to relative density is the clear paste of 1.10 (60 ℃ of mensuration), last activated-charcoal column, earlier use the distilled water eluting, colourless to eluent, 10 times of amounts of reuse, 70% ethanol elution, collect eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with an amount of water for injection; Other gets the Radix Notoginseng total arasaponins of recipe quantity, add an amount of water for injection dissolving, filter, Radix Notoginseng medicinal liquid and Fructus Gardeniae medicinal liquid mixing, regulate pH value to 6.0-7.0,0-4 ℃ of cold preservation 24 hours, 0.45 μ m filter membrane filters, and filtrate adds to the full amount of water for injection, filter with 0.22 μ m filter membrane, the filtrate fill becomes 10ml ampoule injection, 110 ℃ of pressure sterilizings 30 minutes, promptly.Function with cure mainly: removing pathogenic heat from blood and toxic substance from the body, disperse blood stasis and dredge collateral.Cure mainly the cerebral infarction disease.Be used for the apoplexy hemiplegia, hemianesthesia, dizziness and headache, stiff tongue speech is not smoothgoing or in silence, crooked mouth and tongue, mind unconsciousness; Person's falling forward suddenly very, mind is muddle-headed, and a name rale is talked in the nose snore, and dimly red tongue is disturbed on the wind-fire such as stringy and rolling pulse, the card of stasis of blood resistance venation.
Usage and consumption: once-a-day, a 154mg, two weeks of the course of treatment.
Specification: 10ml/ props up, and 154mg is (with jasminoidin, ginsenoside Rg
1, ginsenoside Rb
1, ginsenoside R
1Content sum meter).
Embodiment 2Injection
Fructus Gardeniae 190g Radix Notoginseng total arasaponins 7g
Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder,, collect percolate with 75% ethanol percolation of 7 times of amounts, reclaim ethanol, be concentrated into 65 ℃ of relative densities and be 1.10 clear paste, add 5 times of water gagings dilutions, stir evenly, 0-4 ℃ of cold preservation 49 hours filters, 60 ℃ of relative densities of filtrate decompression simmer down to are 1.10 clear paste, and last activated-charcoal column is used earlier the distilled water eluting, colourless to eluent, 10 times of amounts of reuse, 70% ethanol elution is collected eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with an amount of water for injection; Other gets the Radix Notoginseng total arasaponins of recipe quantity, add an amount of water for injection dissolving, filter, Radix Notoginseng medicinal liquid and Fructus Gardeniae medicinal liquid mixing, regulate pH value to 6.0-7.0,0-4 ℃ of cold preservation 24 hours, 0.45 μ m filter membrane filters, and filtrate adds to the full amount of water for injection, filter with 0.22 μ m filter membrane, the filtrate fill becomes 10ml ampoule injection, and 115 ℃ of pressure sterilizings 30 minutes promptly get injection.Usage and consumption: once-a-day, a 154mg, two weeks of the course of treatment.Specification: 10ml/ props up, and 154mg is (with jasminoidin, ginsenoside Rg
1, ginsenoside Rb
1, ginsenoside R
1Content sum meter).
Embodiment 3Tablet
Fructus Gardeniae 175g Radix Notoginseng 90g.
Get Fructus Gardeniae, the Radix Notoginseng of recipe quantity, be ground into coarse powder, with 75% ethanol percolation of 7 times of amounts, collect percolate, reclaim ethanol, be concentrated into 65 ℃ of relative densities and be 1.10 clear paste, add the dilution of 5 times of water gagings, stir evenly, 0-4 ℃ of cold preservation 49 hours, filter, 60 ℃ of relative densities of filtrate decompression simmer down to are 1.10 clear paste, and often the regulation grain is made granule 100 grams, add excipient, tabletting (the pelletizing press sheet machine is once finished).Every heavy 0.5 gram.
Embodiment 4Capsule
Jasminoidin 70g Radix Notoginseng total arasaponins 60g
Often regulation becomes capsule.
Embodiment 5The method of quality control of present composition injection
Discrimination method: a. gets this product 5ml, and water bath method, residue add dissolve with ethanol and quantitative the commentaries on classics is dissolved in volumetric flask 5ml, and other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 4mg, in contrast product solution; Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2000 VIB); Draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10: 7: 2: 0.5 ethyl acetate-acetone-formic acid-water is developing solvent, launches, and takes out, and dries, and spray is heated with 5% vanillin sulfuric acid solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; B. get this product 10ml, in addition water saturated n-butyl alcohol jolting is extracted 3 times, and each 10ml merges n-butanol extracting liquid, with the saturated water liquid washing of n-butyl alcohol 3 times, each 100ml; Get the n-butanol layer reclaim under reduced pressure to doing, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets ginsenoside Rb
1, the ginsenoside Rg
1And Panax Notoginseng saponin R
1Reference substance adds methanol and makes the mixed liquor that every ml contains 6mg, in contrast product solution; Test according to thin layer chromatography (appendix IIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, in ambient temperature below 20 ℃, lower floor's solution of placing below 10 ℃ with 13: 7: 2 chloroform-methanol-water is developing solvent, launches, take out, dry, spray is with 1 → 10 sulphuric acid ethanol liquid, and it is clear to be heated to speckle colour developing in 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Content assaying method: a. jasminoidin, the photograph high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 D) measure; Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 15: 85 acetonitrile-waters are mobile phase; The detection wavelength is 238nm; Theoretical cam curve is calculated by the jasminoidin peak should be not less than 1500; It is an amount of that the preparation precision of reference substance solution takes by weighing the jasminoidin reference substance, adds methanol and make dissolving in right amount, makes the solution that every 1ml contains 0.144mg, promptly; The preparation of need testing solution, accurate this product 1ml that draws, water bath method, residue adds anhydrous alcohol solution, and ethanol liquid is transferred to another crucible, water bath method, residue is with dissolve with methanol and be transferred in the 100ml measuring bottle, adds methanol to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject high performance liquid chromatograph, measure, promptly; This product contains jasminoidin (C
17H
24O
10) every ml must not be less than 4.7mg; B. arasaponin is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 D); Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With acetonitrile-water binary gradient is mobile phase (seeing Table 1); The detection wavelength is 203nm; Theoretical cam curve is with the ginsenoside Rg
1The peak calculates should be not less than 2500; The reference substance solution preparation, precision takes by weighing arasaponin R1, ginsenoside Rg
1, ginsenoside Rb
1Add dissolve with methanol in right amount, make every 1ml and contain Panax Notoginseng saponin R
11.5mg, the ginsenoside Rg
12.8mg, ginsenoside Rb
14.1mg the mixing reference substance solution; The need testing solution preparation, the accurate sample 5ml that draws extracts three times with the water-saturated n-butanol jolting, each 10ml; N-butanol layer merges, and with the washing of 10ml n-butyl alcohol saturation water once, abandons water layer, the n-butanol layer reclaim under reduced pressure, and to doing, residue, shakes up to 5ml with dissolve with methanol, filters with microporous filter membrane 0.45 μ m, promptly; Algoscopy, the accurate reference substance solution 3 μ l that draw, 7 μ l, need testing solution 10 μ l inject high performance liquid chromatograph, calculate with the external standard two-point method, promptly; The every ml of this product contains Panax Notoginseng saponin R
1Must not be less than 0.2mg; The ginsenoside Rg
1Must not be less than 0.8mg; Ginsenoside Rb
1Must not be less than 2.0mg;
Table 1: acetonitrile-water binary gradient elution table
Time (min) flow velocity (ml/min) A (water) B (acetonitrile)
0 1.0 80 20
20 1.0 60 40
Injection method of quality control of the present invention also can adopt the method for finger printing: the preparation of a. need testing solution: the accurate injection 5ml that draws, extract twice with water saturated n-butyl alcohol jolting, for the first time 10ml; 5nl merges n-butanol extracting liquid for the second time; With the saturated water 10ml backwash of n-butyl alcohol once, discard water layer, the n-butanol extracting liquid reclaim under reduced pressure is to doing, and residue is with dissolve with methanol and be settled in the 5ml volumetric flask, shakes up, and the 0.45um filter membrane filters, promptly; The preparation of reference substance solution: get the jasminoidin reference substance, the solution of making 0.5mg/ml with methanol is product solution in contrast; Assay method: detect wavelength 238nm, the acetonitrile-water gradient elution sees Table 2; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 101, writes down 60 minutes chromatogram, promptly; With the area at the retention time of the chromatographic peak (S peak) of jasminoidin and peak is 1 to calculate relative retention time and peak area ratio; Finger printing and technical parameter; Be 60 minutes writing time; The demarcation (relative retention time) of total fingerprint peaks: it is as follows to demarcate total fingerprint peaks according to the testing result of 10 batches of test samples: 1 (0.404) 2 (0.462) 3 (0.510) 4 (0.589) 5 (0.618) 6 (0.666) 7 (0.722) 8 (0.798) S (1.000) 9 (1.085) 10 (1.110) 11 (1.197) 12 (1.238) 13 (1.277); The ratio of total fingerprint peaks area: 1: S=1: (0.060-0.113); The part peak shape is described: separate when bad, peak 4 is the acromion at peak 5; Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak gross area must not be greater than 5% of peak area;
Instrument reagent high performance liquid chromatograph: Waters2695 pump; Waters 2996 UV-detector; The Millcnnium32 chromatographic work station; Diamonsil C18 post (5 μ m, 4.6mm * 150mm); Acetonitrile is chromatographically pure (U.S. J.T.Baker); The WAHAHA pure water; Methanol is chromatographically pure (Beijing prosperousization Fine Chemical Works);
Table 2 eluent gradient eluting table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
B. the preparation of need testing solution: the accurate injection 10ml that draws, divide the secondary jolting to extract with water saturated n-butyl alcohol 20m1, merge n-butanol extracting liquid; Saturated water 300ml divides three backwashes with n-butyl alcohol, discards water layer, and the n-butanol layer reclaim under reduced pressure is to doing, and residue is with dissolve with methanol and be settled in the 2ml measuring bottle, shakes up, and the 0.45um filter membrane filters, promptly; Preparation according to thing: get the ginsenoside Rg
1Reference substance (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute), the solution of making 2mg/ml with methanol is product solution in contrast; Assay method: detect wavelength 210nm, sensitivity 0.1AUFS: the acetonitrile-water gradient elution sees Table 3; Theoretical cam curve is by the ginsenoside Rg
1Calculate, should not hang down 30000; The accurate need testing solution 25 μ l that draw inject high performance liquid chromatograph, write down 70 minutes chromatogram, promptly; With the ginsenoside Rg
1The retention time of chromatographic peak (S peak) and the area at peak be 1 to calculate relative retention time and peak area ratio; Finger printing and technical parameter; Be 70 minutes writing time; The demarcation (relative retention time) of total fingerprint peaks: it is as follows to demarcate total fingerprint peaks according to the testing result of 10 batches of test samples: 1 (0.877) S (1.000) 2 (1.027) 3 (1.555) 4 (1.592) 5 (1.652) 6 (1.675) 7 (1.708) 8 (1.744) 9 (1.799) 10 (2.005) 11 (2.042) 12 (2.074) 13 (2.111) 14 (2.150) 15 (2.212) 16 (2.235) 17 (2.364) 18 (2.391) 19 (2.420) 20 (2.440) 21 (2.771); The ratio of total fingerprint peaks area: S: 3: 8: 21=1: (0.222-0.412): (0.405-0.735): (1.428-2.380); Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak gross area must not be greater than 5% of the gross area;
Instrument reagent: Waters 2695 Waters 2996 UV-detector Millcnnium32 chromatographic work station Symmetryshild
TMRP
18(5 μ m, 4.6mm * 250mm); Other is analytical pure for Fisher acetonitrile (U.S.'s chromatographically pure) Fisher methanol (U.S.'s chromatographically pure) WAHAHA pure water
Table 3 eluent gradient eluting table
Time (min) flow velocity (ml/min) A-water B-acetonitrile
0 1.0 83 17
20 1.0 76 24
40 1.0 60 40
50 1.0 50 50
55 1.0 0 100
60 1.0 0 100
70 1.0 0 100
Claims (10)
1, a kind of Chinese medicine composition for the treatment of apoplexy is characterized in that this Chinese medicine composition made by following raw material medicaments:
Fructus Gardeniae 150-220 weight portion Radix Notoginseng 70-110 weight portion.
2, Chinese medicine composition as claimed in claim 1 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Fructus Gardeniae 170-200 weight portion Radix Notoginseng 80-100 weight portion.
3, Chinese medicine composition as claimed in claim 1 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Fructus Gardeniae 150-220 weight portion Radix Notoginseng total arasaponins 3-9 weight portion
4, Chinese medicine composition as claimed in claim 3 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Fructus Gardeniae 170-200 weight portion Radix Notoginseng total arasaponins 5-7 weight portion.
5, Chinese medicine composition as claimed in claim 4 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Fructus Gardeniae 175 weight portion Radix Notoginseng total arasaponinss 5.5 weight portions.
6, Chinese medicine composition as claimed in claim 1 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Jasminoidin 3.8-11.5 weight portion Radix Notoginseng total arasaponins 3-9 weight portion.
7, Chinese medicine composition as claimed in claim 6 is characterized in that this Chinese medicine composition made by following raw material medicaments:
Jasminoidin 6.3-8.9 weight portion Radix Notoginseng total arasaponins 5-7 weight portion.
8, preparation method as the described Chinese medicine composition composition injection of claim 3-5, it is characterized in that this method is: get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, the 65-80% ethanol percolation of doubly measuring with 7-9, collect percolate, reclaim ethanol, the clear paste that to be concentrated into 55-70 ℃ of relative density be 1.00-1.20 adds 3-5 times of water gaging dilution, stir evenly, 0-4 ℃ cold preservation 45-50 hour, filter the clear paste that 55-70 ℃ of relative density of filtrate decompression simmer down to is 1.00-1.20, last activated-charcoal column, earlier use the distilled water eluting, colourless to eluent, reuse 9-11 doubly measures the 65-80% ethanol elution, collect eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with an amount of water for injection; Other gets the Radix Notoginseng total arasaponins of recipe quantity, add an amount of water for injection dissolving, filter, Radix Notoginseng medicinal liquid and Fructus Gardeniae medicinal liquid mixing, regulate pH value to 5.5-7.5,0-4 ℃ cold preservation 20-30 hour, 0.45 μ m filter membrane filters, filtrate adds to the full amount of water for injection, filter with 0.22 μ m filter membrane, the filtrate fill becomes 10ml ampoule injection, 110-120 ℃ pressure sterilizing 20-40 minute, promptly.
9, the preparation method of Chinese medicine composition composition injection as claimed in claim 8, it is characterized in that this method is: get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, with 70% ethanol percolation of 8 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of relative densities and be 1.10 clear paste, add 4 times of water gagings dilutions, stir evenly, 0-4 ℃ of cold preservation 48 hours filters, and 60 ℃ of relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, earlier use the distilled water eluting, colourless to eluent, 10 times of amounts of reuse, 70% ethanol elution, collect eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with an amount of water for injection; Other gets the Radix Notoginseng total arasaponins of recipe quantity, add an amount of water for injection dissolving, filter, Radix Notoginseng medicinal liquid and Fructus Gardeniae medicinal liquid mixing, regulate pH value to 6.0-7.0,0-4 ℃ of cold preservation 24 hours, 0.45 μ m filter membrane filters, and filtrate adds to the full amount of water for injection, filter with 0.22 μ m filter membrane, the filtrate fill becomes 10ml ampoule injection, 110 ℃ of pressure sterilizings 30 minutes, promptly.
10. as the application of the described Chinese medicine composition of claim 1-7 in the medicine of preparation treatment apoplexy.
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|---|---|---|---|
| CN 03149526 CN1253169C (en) | 2003-06-19 | 2003-07-15 | Chinese medicine composition for curing apoplexy and its preparation method |
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|---|---|---|---|
| CN03137402.6 | 2003-06-19 | ||
| CN03137402 | 2003-06-19 | ||
| CN 03149526 CN1253169C (en) | 2003-06-19 | 2003-07-15 | Chinese medicine composition for curing apoplexy and its preparation method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1330333C (en) * | 2005-06-09 | 2007-08-08 | 北京中医药大学 | Chinese medicinal composition and related preparations thereof |
| CN109172624A (en) * | 2018-11-04 | 2019-01-11 | 上海博桂文化传播有限公司 | A kind of pharmaceutical composition and its application for treating cerebral arterial thrombosis |
| CN114425029A (en) * | 2022-02-14 | 2022-05-03 | 西安绿天生物技术有限公司 | Plant extract composition with anti-inflammatory and allergy-relieving functions, preparation method and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1305493C (en) * | 2005-06-09 | 2007-03-21 | 北京中医药大学 | Chinese medicine effective part for treating cerebral apoplexy and its separation preparing process |
-
2003
- 2003-07-15 CN CN 03149526 patent/CN1253169C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1330333C (en) * | 2005-06-09 | 2007-08-08 | 北京中医药大学 | Chinese medicinal composition and related preparations thereof |
| CN109172624A (en) * | 2018-11-04 | 2019-01-11 | 上海博桂文化传播有限公司 | A kind of pharmaceutical composition and its application for treating cerebral arterial thrombosis |
| CN114425029A (en) * | 2022-02-14 | 2022-05-03 | 西安绿天生物技术有限公司 | Plant extract composition with anti-inflammatory and allergy-relieving functions, preparation method and application thereof |
| CN114425029B (en) * | 2022-02-14 | 2024-05-10 | 西安绿天生物技术有限公司 | Plant extract composition with anti-inflammatory and allergy relieving effects, preparation method and application thereof |
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| Publication number | Publication date |
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| CN1253169C (en) | 2006-04-26 |
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