CN101474220A - Novel use of total flavone of effective component of Cotinus coggygria and preparation - Google Patents
Novel use of total flavone of effective component of Cotinus coggygria and preparation Download PDFInfo
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Abstract
One of the tasks of the invention is to define an effective part for using cotinus coggygria, and provide pharmacodynamics usages of the effective part and various preparations. The new usage of the effective part of the cotinus coggygria is that the leaves, stems, branches and roots of the cotinus coggygria are collected to extract general flavone as a drug which can improve animal organism hypoxia tolerance or thrombolysis, etc. The invention has the advantages that the effective part of the cotinus coggygria is defined, and the effective components are separated for medical use, thus obviously improving the therapeutic effect. Experiments prove that the preparation of the invention can obviously improve animal hypoxia tolerance and the effect thereof is equal to a western medicine antianginal drug Propranolol, thus avoiding the toxic side effect of the western medicine and having the actions of improving blood circulation and reducing whole blood viscosity. Therefore, the preparation can be applicable for preventing and curing coronary heart disease, angina pectoris, miocardial infarction, as well as obstruction of Qi in chest, dizziness, brachypnea, palpitation, chest distress or thoracodynia caused by internal resistance of congestion. Meanwhile, the preparation of the invention also has the actions of anticoagulation in vitro and thrombus dissolving.
Description
Technical field
The present invention relates to new purposes and preparation that a kind of human drugs technical field, particularly effective component of Cotinus coggygria are total flavones.
Background technology
Ramulus et Folium Cotini Coggygiae (Cotinus coggygra Scop.) is the Anacardiaceae fuste, and machaka or dungarunga reach as high as 8m, and autumn is red through white posterior lobe complexion changed, is commonly called as the Fragrance Hill red autumnal leaves.Though this plant does not also enter China's pharmacopeia as medical material, many in recent years pharmacodynamics tests show that it has antibacterial action, significant antihepatitic activity and obvious anti-fatigue effect etc., and therefore, Ramulus et Folium Cotini Coggygiae has been carried out a large amount of research by people as medicine.The inventor has studied the effect of Ramulus et Folium Cotini Coggygiae aspect anticoagulation, antithrombotic, hypotensive activity and medicine thereof in Chinese patent application 200610042000.3.
But people still are in the junior stage to the research of Ramulus et Folium Cotini Coggygiae, both concrete effective site of Ramulus et Folium Cotini Coggygiae and medicinal effective ingredient are not carried out deep research, also to the not exploitation of its more drug effect.
Summary of the invention
One of task of the present invention is to determine the medicinal effective site of Ramulus et Folium Cotini Coggygiae, and the pharmacodynamics usages and the various preparation of this effective site are provided.
The new purposes of effective component of Cotinus coggygria, it is to gather FOLIUM ET RAMULUS COTINI, stem, branch and root extraction total flavones as the medicine that improves effects such as animal body resisting oxygen lack or thrombus.
Effective site is that total flavones is as prevention and treatment coronary heart disease, the medicine of angina pectoris or myocardial infarction in the Ramulus et Folium Cotini Coggygiae.
Effective site is the medicine of total flavones as prevention and treatment cerebral thrombosis and thrombotic disease in the Ramulus et Folium Cotini Coggygiae.
Isolate fisetin and Quercetin in the described total flavone of effective component of Cotinus coggygria as medicinal ingredient.
Improve the pharmaceutical preparation of animal body resisting oxygen lack, it contains isolates fisetin and Quercetin as effective ingredient in the total flavone of effective component of Cotinus coggygria.
Pharmaceutical preparation with thrombus effect, it contains isolates fisetin and Quercetin as effective ingredient in the total flavone of effective component of Cotinus coggygria.
The pharmaceutical preparation of described raising animal body resisting oxygen lack, it to be to isolate fisetin and Quercetin as effective ingredient in the total flavone of effective component of Cotinus coggygria, add pharmaceutically acceptable carrier or complementary composition and be prepared from.
Described pharmaceutical preparation with thrombus effect, it to be to isolate fisetin and Quercetin as effective ingredient in the total flavone of effective component of Cotinus coggygria, add pharmaceutically acceptable carrier or complementary composition and be prepared from.
Advantage of the present invention is an effective site of having determined Ramulus et Folium Cotini Coggygiae, and isolate effective ingredient as medicinal, obviously improved therapeutic effect, prove by experiment, preparation of the present invention can significantly improve the animal hypoxia-bearing capability, effect is suitable with Western medicine anti-anginal drug Propranolol, avoided the toxic and side effects of Western medicine, and has a blood circulation promoting and blood stasis dispelling, reduce the effect of whole blood viscosity, be used for prevention and treatment coronary heart disease, angina pectoris, myocardial infarction and congestion internal resistance the thoracic obstruction, dizzy, breathe hard, cardiopalmus, uncomfortable in chest or chest pain.Simultaneously, preparation of the present invention also has the effect of external anticoagulation and thrombus dissolving.
The specific embodiment
Embodiment 1: isolating total flavones adding starch or dextrin are made granule according to existing technology in the Ramulus et Folium Cotini Coggygiae, are used for the treatment of coronary heart disease, angina pectoris and myocardial infarction.
Embodiment 2: the total flavones of embodiment 1 is made capsule, as the medicine of prevention and treatment cerebral thrombosis and thrombotic disease.
Embodiment 3: with the Ramulus et Folium Cotini Coggygiae total flavones, further separate, obtain Quercetin and fisetin, make injection respectively after being dissolved in solvent, injection is used for the treatment of the first aid medicine of coronary heart disease, angina pectoris and myocardial infarction and the first aid medicine of cerebral thrombosis and thrombotic disease.
In the injection preparation, need to add conventional PH regulator (hydrochloric acid, sodium bicarbonate), antibacterial, osmotic pressure regulator and antioxidant etc.
The Ramulus et Folium Cotini Coggygiae preparation that the relevant pharmacodynamics report proof of preparation of the present invention utilizes the present invention to prepare has the animal body resisting oxygen lack of raising and thrombus effect, specifically sees following adnexa.
Adnexa:
The effective site of Ramulus et Folium Cotini Coggygiae anticoagulant active, collecting season and effective site screening.
1 material and method
1.1 animal
Dog, body weight 15～30kg, male and female are not limit, provide by Xi'an Jiaotong University Medical College's Experimental Animal Center, 26 ± 3 ℃ of Animal House room temperatures, every day artificial lighting 12h, 0.5h respectively takes a breath morning and afternoon every day.The ICR mice, body weight 18～22g, ♀ ♂ half and half, the animal quality quality certification number is provided by Xi'an Jiaotong University Medical College's Experimental Animal Center: the moving card of Shan doctor word 08-004 number, temperature, illumination and ventilation are described with the front, by the sex sub-cage rearing, freely drink water, feed pellet.
1.2 instrument
The MP200B electronic balance, the flat instrument and meter company limited of last current chart; The HHS electric-heated thermostatic water bath, Shanxi Province's hydrology medical apparatus and instruments factory; AE240S type electronic analytical balance, Metler-Toledo Group; 752 type ultraviolet grating spectrophotometers, Shanghai the 3rd analytical tool factory; D101-2 type drying baker, Chinese Shanghai experimental apparatus head factory; CQ-120 type ultrasonic washing unit, the Shanghai medical optical instrument factory of making a leapleap forward; Motic B1-220ALED type biological microscope, Maike Aodi Industry Group Co Ltd; SENCO-R series rotary evaporator, Shensheng Science ﹠ Tech. Co., Ltd., Shanghai; SHZ-D (III) circulation ability of swimming vacuum pump, the Ying Yu of Gongyi City gives magnificent instrument plant; Stopwatch etc.
1.3 test medication
1.3.1 medicine fisetin (Fisetin) reference substance, the bright plant development in science and technology of Shaanxi rising sun company limited, lot number 060610; Quercetin (Quercetin) chemical reference substance, Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 100081-200406; Heparin sodium injection, Jiangsu Wanbang Biological Pharmaceutical Co., Ltd., lot number 050612 faces with the preceding distilled water diluting of using, and regulates its pH=7 with PBS (pH=7.4) solution; Aspirin Enteric-coated Tablets, Bailu Pharmaceutical Co., Ltd., Shaanxi, lot number 070335 faces with preceding and is made into desired concn with 0.5%CMC-Na solution, and regulates its pH=7 with PBS (pH=7.4) solution; Pentobarbital sodium inj, Tianjin gold credit aminoacid company limited, lot number 060609; Ketaject injection, the Shanghai first biochemical Pharma Inc., lot number 060910; Potassium dihydrogen phosphate, sodium hydroxide and sodium chloride etc. are analytical pure.
1.3.2 the preparation at each position of Ramulus et Folium Cotini Coggygiae
1.3.2.1 Ramulus et Folium Cotini Coggygiae stem, branch, leaf and root water extract
(lot number is respectively 060909,060924 and 061117 for the stem of three batches of Ramulus et Folium Cotini Coggygiae medical materials, branch, Ye Jigen, be called first, second batch and the 3rd batch of each position), lot number is dead leaf on 061226 the tree, a tree wet leaves and tree cured leaf down down, all pick up from mountain range, the geographic Qinling Mountains, Baoji, Shaanxi province, professor Guo Zengjun through Xi'an Jiaotong University Medical College is accredited as each position that the Anacardiaceae Ramulus et Folium Cotini Coggygiae belongs to Ramulus et Folium Cotini Coggygiae, wherein first and second batch leaf is green, and the 3rd batch of medical material leaf is red.More than the preparation of each position water extract: get each several part 30g respectively, adding distil water is an amount of, soaks, with electric jacket heating extraction three times, each 1h, merge extractive liquid,, be evaporated to 60mL, put coldly, add 95% ethanol and make it contain alcohol amount to reach 80%, standing over night, filter, behind the decompression recycling ethanol to cumulative volume be 30mL, separately concentration of aqueous solution be 1mg/mL, standby.
1.3.2.2 Ramulus et Folium Cotini Coggygiae ethanol extract, post thing preparation excessively
Stem branch (061117): utilize Ramulus et Folium Cotini Coggygiae stem branch Orthogonal experiment results, carry out Ramulus et Folium Cotini Coggygiae stem branch extracts active ingredients, be about to Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol, 12 times of amounts, pH is 9,1h/ time, extract twice, and get Ramulus et Folium Cotini Coggygiae stem branch alcohol extract, behind the concentrating under reduced pressure, water bath method gets extractum.Get an amount of extractum and be dissolved in the Diluted Alcohol, behind the last macroporous resin, use 15BV
0(column volume) 95% ethanol elution, with the eluent concentrating under reduced pressure, the drying with water bath concentrated solution gets 95% ethanol post eluate.Face with before getting the former an amount of extractum, to desired concn, get an amount of extractum of the latter, be formulated into desired concn with 1.0% sodium carboxymethyl cellulose (CMC-Na) solution with dissolved in distilled water.
Leaf (061117): utilize the FOLIUM ET RAMULUS COTINI Orthogonal experiment results, carry out the FOLIUM ET RAMULUS COTINI extracts active ingredients, be about to FOLIUM ET RAMULUS COTINI 70% ethanol, 20 times of amounts, pH is 5,2h/ time, extracts twice, merging filtrate promptly gets the FOLIUM ET RAMULUS COTINI alcohol extract, behind the concentrating under reduced pressure, water bath method gets 70% ethanol extracted extract, face with before getting an amount of extractum, arrive desired concn with dissolved in distilled water.FOLIUM ET RAMULUS COTINI the water extracted immersing paste preparation process is: the leaf coarse powder adds 12 times of water gagings, water-bath reflux, extract, 2 times, each 1.5h, merging filtrate promptly gets FOLIUM ET RAMULUS COTINI water extract, behind the concentrating under reduced pressure, and water bath method, get its water extracted immersing paste, face, arrive desired concn with dissolved in distilled water with before getting an amount of extractum.
1.4 test method
1.4.1 determination of total flavonoids
1.4.1.1 the preparation of fisetin standard curve [4]
Claim to be dried to the fisetin reference substance 0.0012g of constant weight, with analytical pure 95% dissolve with ethanol and be settled to 100mL, therefrom take out 5mL, 4mL, 3mL respectively, 2mL, 1mL, 0mL put in the 5mL measuring bottle, behind 95% ethanol dilution and standardize solution, measure its trap at λ=367 ± 1nm place, the drawing standard curve gets regression equation.
1.4.1.2 determination of total flavonoids in each position water extract of Ramulus et Folium Cotini Coggygiae
Get each position water extract-alcohol precipitation liquid 5mL of Ramulus et Folium Cotini Coggygiae 1mg/mL respectively, put in the evaporating dish respectively, water-bath is waved near and is done, with a small amount of 95% dissolve with ethanol and be transferred in the 25mL measuring bottle, continue to measure its trap at λ=367 ± 1nm place, calculate and measure content of total flavone in the liquid with behind 95% ethanol dilution and the standardize solution, and then content of total flavone in each medicinal part of calculating Ramulus et Folium Cotini Coggygiae
And tabulation relatively.
1.4.1.3 application of sample recovery test
Getting lot number and be 061117 Ramulus et Folium Cotini Coggygiae stem water extract 0.1mL puts in the 5mL volumetric flask, adding concentration again is 95% alcoholic solution 0.06mL of 0.464mg/mL fisetin, and mixing is settled to 5mL with 95% ethanol, measure its trap at λ=367 ± 1nm place, measure the response rate.
1.4.2 external anticoagulation of dog and thrombolytic test [5]
1.4.2.1 anticoagulation experiment: dog, after ketalar and sodium phenobarbital intramuscular anesthesia, operation, the common carotid artery intubate is got blood.Get 12 sterilization small test tubes, add 0.3mL 0.9% NaCl solution respectively; 0.3mL the heparin sodium aqua of 5.556U/mL; 0.3mL the heavy dose group solution (1mg/mL) of second batch and the 3rd batch stem of Ramulus et Folium Cotini Coggygiae, branch, Ye Hegen, Quercetin heavy dose of group solution (0.01mg/mL) and the heavy dose of group of fisetin solution (0.01mg/mL).And then add 0.7mL blood at every pipe, and the mixing that turns upside down is put 37 ℃ of waters bath with thermostatic control, and every 30min observes, and record is taken pictures.Behind the 4h, each test tube of jolting is got top blood and slime smear gently, and microscopically is observed red cell morphology and taken pictures.Other experiment conditions are constant, in further observing, low dose of Ramulus et Folium Cotini Coggygiae each several part solution (0.5mg/mL, 0.25mg/mL), Quercetin and fisetin (0.005mg/mL, 0.0025mg/mL) solution is to the influence of the external clotting time of dog.More than each solution face with preceding be that to regulate its pH be 7 for 7.4 PBS with pH.
1.4.2.2 the thromboembolism experiment: the dog common carotid artery is got blood.Get 12 sterilization small test tubes, after every pipe adds 1mL blood, treats blood coagulation, add 1mL 0.9% NaCl respectively; The heparin sodium aqua of 1mL 5.556U/mL; The heavy dose group solution (1mg/mL) of second batch and the 3rd batch stem of each 1mL Ramulus et Folium Cotini Coggygiae, branch, Ye Hegen, Quercetin heavy dose of group solution (0.01mg/mL) and the heavy dose of group of fisetin solution (0.01mg/mL).And then the mixing that turns upside down 1 time, put 37 ℃ of waters bath with thermostatic control, every 10min, every the test tube that turns upside down three times enters the thrombosis internal-response to promote each solution, and every 30min observes, and record is taken pictures.Behind the 4h, removal of thromboses, filter paper claim weight in wet base after blotting surplus blood, simultaneously, get the liquid smear of pulling out behind the thrombosis, at microscopically, observe hemocyte release conditions and form, and take pictures.Other experiment conditions are constant, in further observing, low dose of Ramulus et Folium Cotini Coggygiae each several part solution (0.5mg/mL, 0.25mg/mL), Quercetin and fisetin (0.005mg/mL, 0.0025mg/mL) solution is to the influence of molded thrombosis.
1.4.2.3 the experiment of thromboembolism rate: the dog common carotid artery is got blood.The preparation experiment thrombosis cuts into about equally fritter of size with thrombosis, and every heavily about 0.4g gets 6, and adding has been placed with 0.9% the NaCl solution of 0.6mL successively; 0.6mL the heparin sodium aqua of 5.556U/mL; 12 clean sterilization test tubes of the heavy dose group solution (2mg/mL) of second batch and the 3rd batch stem of each 0.6mL Ramulus et Folium Cotini Coggygiae, branch, Ye Hegen, Quercetin heavy dose of group solution (0.02mg/mL) and fisetin heavy dose of group solution (0.02mg/mL).Put 37 ℃ of waters bath with thermostatic control then, take out all test tubes behind the 2h, careful removal of thromboses, and after blotting the thrombosis surface moisture gently with the filter paper bar, weigh calculating thromboembolism rate successively.Other experiment conditions are constant, in further observing, low dose of Ramulus et Folium Cotini Coggygiae each several part solution (1.0mg/mL, 0.5mg/mL), Quercetin and fisetin (0.01mg/mL, 0.005mg/mL) solution is to the influence of the external molded thromboembolism rate of dog.
Dissolution rate=(W-G)/W * 100%
W: thrombus weight before the experiment
G: experiment back thrombus weight
1.4.3 anticoagulation test in the mice body
1.4.3.1 37 ℃ of thermostatic water-bath methods
The ICR mice, body weight 18～22g, ♀, ♂ half and half, by the body weight layering, random packet: control group administered physiological saline 0.18g/kg, aspirin group 0.1g/kg, the heavy dose of group of the water extract of Ramulus et Folium Cotini Coggygiae stem branch (061117) 70% ethanol extraction, alcohol extraction residue, 95% ethanol post eluate, water extract and FOLIUM ET RAMULUS COTINI 70% ethanol extraction and its water extract (2.8g/kg), the heavy dose of group of Quercetin and fisetin (0.0224g/kg), each group is irritated stomach 0.4ml/20g.Reference literature [5], behind fasting 12～16h, each group is irritated the normal saline of stomach with volume respectively, aspirin 1% sodium carboxymethyl cellulose (CMC-Na) suspension, each heavy dose of group solution of Ramulus et Folium Cotini Coggygiae.Irritated behind the stomach 57～59 minutes, mice is placed holder, make its afterbody vertical, 60min behind the filling stomach, cut off at distance tail point 1.5mm place, and (behind small beaker dress normal saline, be put in the water-bath) in the normal saline with 37 ℃ ± 0.5 ℃ of tail point immersion, record is cut off the tail point to stopping hemorrhage interval, is the bleeding time.Other experiment conditions are constant, further observe and write down the bleeding time of Ramulus et Folium Cotini Coggygiae each several part small dose group (0.7g/kg) solution, Quercetin and fisetin small dose group (0.0056g/kg) solution.
1.4.3.2 ice-water bath method
The same thermostatic water-bath method of basic operation, difference are before the administration, to have carried out the screening of mice bleeding time, if the preceding bleeding time of administration greater than the mice of 10min, then eliminates; Test is to carry out in ice-water bath simultaneously.Utilize the ice-water bath method to negative control group normal saline 0.18g/kg, positive controls aspirin 0.1g/kg, Ramulus et Folium Cotini Coggygiae stem branch (061117) 70% ethanol extraction, 95% ethanol post eluate, water extract, the heavy dose of group of FOLIUM ET RAMULUS COTINI water extract (2.8g/kg) and small dose group (0.7g/kg), the bleeding time of the heavy dose of group of fisetin (0.0224g/kg), small dose group (0.0056g/kg) is measured.
2 results and conclusion
2.1 determination of total flavonoids
2.1.1 the preparation of fisetin standard curve
2.1.1.1 measure the trap of each concentration of fisetin reference substance at λ=367 ± 1nm place, must the standard curve regression equation be: Y=0.1303X+0.028, R
2=0.9991 (n=6) is when fisetin concentration concerns good in 3～12 micrograms/mL scope internal linear.
2.1.2 determination of total flavonoids in each position water extract of Ramulus et Folium Cotini Coggygiae
Three batches of Ramulus et Folium Cotini Coggygiae stems, branch, leaf, root and lot numbers be 061226 Ramulus et Folium Cotini Coggygiae tree go up dead leaf, tree down wet leaves and tree down the contained separately determination of total flavonoids of cured leaf the results are shown in Table 1, Fig. 1 and table 2.Table 1 as seen, total flavones is compared with stem separately in the branch of first, second batch medical material, leaf and the root, there were significant differences (p＜0.01); Total flavones is compared with its stem in branch, leaf and the root of the 3rd batch of medical material, variant (p＜0.05); Fig. 1 as seen, along with increasing season, general flavone content is in rising trend in Radix cotini pubescentis, branch, stem and the leaf, wherein with general flavone content in the leaf for the highest; Table 2 as seen, in the 3rd batch of leaf total flavones and tree go up dead leaf, tree down cured leaf and tree down wet leaves compare, significant difference (p＜0.01) is arranged, tree down cured leaf and tree down in the wet leaves general flavone content and tree upward the total flavones in the dead leaf compare, significant difference (p＜0.01) is arranged.Conclusion: if be foundation with the general flavone content, the medicinal part of Ramulus et Folium Cotini Coggygiae is leaf, and its collecting season is good for the November leaf color that reddens.
Determination of total flavonoids (n=5) in each position of table 1 Ramulus et Folium Cotini Coggygiae different batches crude drug
Annotate: " * " expression is compared variant (p＜0.05) with the stem general flavone content; " * * " expression is compared with the stem general flavone content, and significant difference (p＜0.01) is arranged.
Table 2 Ramulus et Folium Cotini Coggygiae tree goes up dead leaf, tree cured leaf and tree determination of total flavonoids (n=5) in the wet leaves down down
Annotate: " * * " expression goes up with tree that general flavone content relatively has significant difference (p＜0.01) in the dead leaf; General flavone content relatively has significant difference (p＜0.01) in " Δ Δ " expression and the 3rd batch of leaf.
2.1.1.3 the application of sample recovery test the results are shown in Table 3.The result is as seen: this method is feasible.
The application of sample recovery test (n=5) of table 3 Ramulus et Folium Cotini Coggygiae stem
2.2 external anticoagulation of dog and thrombolytic test
Anticoagulation test: with the negative contrast of normal saline (0.9%NaCl), with the positive contrast of heparin sodium, respectively carry out the anticoagulation experiment with Ramulus et Folium Cotini Coggygiae, found that: after each group begins to form thrombosis, hatch at 37 ℃ of constant temperature, along with moisture oozes out thrombosis atrophy gradually, but finally all there is thrombosis to exist, the thrombosis maximum that the normal saline group forms, what the thrombosis minimum that the heparin sodium positive controls forms had does not almost have, each administration group of Ramulus et Folium Cotini Coggygiae and Quercetin and fisetin group, the thrombosis size of formation (is seen Fig. 2, Fig. 4) between normal saline group and heparin sodium group; The clotting time of the big or middle dosage group of each administration group of Ramulus et Folium Cotini Coggygiae is compared with the normal saline group all has significant difference (p＜0.01), and the thrombus formation time between each position of each batch of Ramulus et Folium Cotini Coggygiae does not have significant difference (p〉0.05) (seeing Table 1).Experimental result shows that each extractive part of Ramulus et Folium Cotini Coggygiae has tangible blood coagulation resisting function.
Each extractive part of table 1 Ramulus et Folium Cotini Coggygiae, Quercetin and fisetin to the external clotting time of dog, thrombotic influence (± s)
Annotate: compare * * P＜0.01, * P＜0.05 with the normal saline group.
Behind the 4h, each test tube of jolting is got top blood and slime smear gently, and microscopically is observed red cell morphology and taken pictures to observe and find: the negative control group cell is many to be arranged with the money shape that vanishes, and becomes block; Heparin sodium group erythrocyte is more, and cellular morphology is good, is dispersed in substantially, is biconcave disc; The red cell morphology rounding of each of Ramulus et Folium Cotini Coggygiae batch each position group, be dispersed in, wherein the erythrocyte number of root portion is less; The red cell morphology of Quercetin and fisetin group also rounding, be dispersed in, wherein the cell number of fisetin group is obviously more than the Quercetin group.The result shows that all there is significant anticoagulant active at each position of Ramulus et Folium Cotini Coggygiae, and the anticoagulation effective ingredient is two kinds of main component-fisetins and the Quercetin in its contained total flavones.
The A normal saline; The B heparin sodium; C-F is respectively dosage (0.5mg/mL) in stem, branch, leaf and the root of Ramulus et Folium Cotini Coggygiae (060924); G-J is respectively dosage (0.5mg/mL) in Ramulus et Folium Cotini Coggygiae (061117) stem, branch, leaf and the root; K, L are respectively dosage group in Quercetin and the fisetin (5 μ g/mL).
2.2 the thromboembolism of Ramulus et Folium Cotini Coggygiae
With the negative contrast of normal saline (0.9%NaCl), the positive contrast of heparin sodium is carried out the experiment of dog thromboembolism with each position of Ramulus et Folium Cotini Coggygiae, and 37 ℃ of constant temperature are hatched the 2h discovery: the thrombosis in the negative control group test tube is bigger, top flat; The thrombosis of positive controls is tiny, is difficult for differentiating the top, contains the more erythrocyte that dissolving discharges from thrombosis in the supernatant; The thrombi size is between negative, positive group in each position of Ramulus et Folium Cotini Coggygiae and Quercetin, the fisetin, and supernatant part color is darker, and more erythrocyte is arranged, and each organizes the thrombosis top phenomenon of splaying substantially, promptly dissolve phenomenon, and topside area is less than negative control group.The result shows that each extractive part of Ramulus et Folium Cotini Coggygiae, Quercetin and fisetin have thrombolytic effect.
Behind the 3h, removal of thromboses, filter paper claims weight in wet base after blotting surplus blood, simultaneously, get the liquid smear of pulling out behind the thrombosis, at microscopically, observe hemocyte release conditions and form, find (1): dog 1mL blood is after external the solidifying and after each administration group effect, thrombosis diminishes substantially, each position of Ramulus et Folium Cotini Coggygiae, Quercetin big, the heavy dose group of middle dosage group and fisetin is compared with the normal saline group to the dissolution of molded thrombosis and significant difference is all arranged (p＜0.01), the middle dosage group of fisetin and normal saline group thrombolytic effect variant (p＜0.05), compare respectively at red autumnal leaves Ramulus et Folium Cotini Coggygiae and each position of greenery Ramulus et Folium Cotini Coggygiae, and its thrombolytic effect does not have significant difference (p〉0.05) (seeing Table 1); (2): in the blood smear, negative experimental group erythrocyte number is few, and form is good; The positive controls cell is maximum, and form is normal; In the experimental group behind the second batch of medical material root extract thrombolytic erythrocyte number and negative group quite, other respectively organizes cell obviously more than feminine gender group (seeing Fig. 8 A-L).The result shows that each extractive part of Ramulus et Folium Cotini Coggygiae, fisetin and Cortex querci dentatae have thrombolytic effect, and Quercetin and fisetin may be the effective ingredient of Ramulus et Folium Cotini Coggygiae thrombolytic.
2.3 with the dog fresh thrombus is material, organizes medicine under 37 ℃ of constant temperatures with each, calculates thromboembolism rate (seeing Table 2) behind the effect 2h.The result is as seen: normal saline has dissolution to thrombosis; Heparin sodium, each extractive part of Ramulus et Folium Cotini Coggygiae, Quercetin and the big or middle dosage group of fisetin are compared with negative control group, and thrombosis is had significant dissolution (p＜0.01); Ramulus et Folium Cotini Coggygiae root extract small dose group and negative control group compare, and its dissolution rate to thrombosis does not have difference (p＜0.05); Between each position of Ramulus et Folium Cotini Coggygiae and the thromboembolism rate between red autumnal leaves Ramulus et Folium Cotini Coggygiae and the greenery Ramulus et Folium Cotini Coggygiae do not have difference (p＜0.05).The result shows: Ramulus et Folium Cotini Coggygiae has thrombolytic effect; The thrombolytic effect at red autumnal leaves Ramulus et Folium Cotini Coggygiae and each position of yellow leaf Ramulus et Folium Cotini Coggygiae does not have difference; Quercetin and fisetin also have the thrombus dissolving effect, and they are effective ingredient of Ramulus et Folium Cotini Coggygiae thrombus dissolving.
Each position of table 2 Ramulus et Folium Cotini Coggygiae, Quercetin and fisetin to the influence of the molded thromboembolism rate of dog (%, ± s)
Annotate: compare * * P＜0.01, * P＜0.05 with the normal saline group.
2.4 anticoagulation test in the mice body
2.4.1 37 ℃ of thermostatic water-bath methods: the bleeding time of aspirin group mice compares with the normal saline group, property that there were significant differences (p＜0.01); The large and small dosage group of Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol extraction, the heavy dose of group of Ramulus et Folium Cotini Coggygiae stem branch water extract, and the bleeding time of the heavy dose of group of fisetin mice compare with the normal saline group, variant (p＜0.05), and these four dosage groups compare with the aspirin group to the bleeding time of mice, do not have difference (p〉0.05).The result shows: Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol extraction and water extract thereof, fisetin all have blood coagulation resisting function, and the aspirin of its anticoagulant effect and 0.1g/kg is suitable.
The relevant extract of table 3 Ramulus et Folium Cotini Coggygiae, Quercetin and fisetin to the influence in mice bleeding time (± s)
Annotate: compare with the normal saline group, " * * " represents P＜0.01, and " * " represents P＜0.05; Compare " Δ Δ " expression P＜0.01, " Δ " expression P＜0.05 with the aspirin group.
2.4.2 ice-water bath method: owing to using 37 ℃ of thermostatic water-bath methods to measure in the process in mice bleeding time, the individual difference of finding mice is very big, diversity is bigger, the diversity that causes the mice bleeding time increases, so design uses the ice-water bath method that the gained bleeding time in 37 ℃ of thermostatic water-bath methods is reaffirmed the influence in mice bleeding time greater than the dosage group of 7min.The result is as seen: the anticoagulant effect and the normal saline group variant (p＜0.05) of the heavy dose of group of Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol extraction, the 95% ethanol post eluate of Ramulus et Folium Cotini Coggygiae stem branch and fisetin; Except that crossing Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol extraction, Ramulus et Folium Cotini Coggygiae stem branch water extract, fisetin small dose group separately, all the other each sample dose groups are compared with the aspirin group the influence in mice bleeding time, all do not have diversity (p〉0.05).The result shows: 95% ethanol elution of gained part (total flavones) behind the Ramulus et Folium Cotini Coggygiae stem branch 70% ethanol extraction solution upper prop, have anticoagulant effect, and the aspirin of anticoagulant effect and 0.1g/kg is suitable, and its anticoagulant active composition is wherein contained fisetin.
The relevant extract of table 4 Ramulus et Folium Cotini Coggygiae, Quercetin and fisetin to the influence in mice bleeding time (± s)
Embodiment two
1 method
The experiment of mice anoxia enduring
[11]: get 80 of ICR mices, male and female half and half are divided into 8 groups at random by body weight, 10 every group.Be respectively: (1), normal group (irritating the stomach distilled water); (2), Propranolol group: (0.0135g/kg); (3), the large, medium and small dosage group of Ramulus et Folium Cotini Coggygiae (20.0g/kg, 10.0g/kg, 5.0g/kg); (4), the large, medium and small dosage group of compound recipe Ramulus et Folium Cotini Coggygiae (20.0g/kg, 10.0g/kg, 5.0g/kg).Irritate hello administration, irritate the long-pending 10mL/kg of body of stomach, behind the 1h, mice is put into the 125mL tool plug wide mouthed bottle that the 7.5g sodium carbonate is housed respectively, vaseline is smeared bottleneck, sealing, and the time of mice to anoxia asphyxia death is observed and writes down in timing.Finish one and take turns back exchange bottle clockwise.
2 results and conclusion
Ramulus et Folium Cotini Coggygiae sees Table 1 to the influence of mice anoxia enduring.Table 1 as seen, with negative control group relatively, the heavy dose of group of effective component of Cotinus coggygria has the resisting oxygen lack (P＜0.001) of highly significant, middle dosage group has remarkable resisting oxygen lack (P＜0.05), low dose group does not have resisting oxygen lack (P〉0.05).
Table 1 Ramulus et Folium Cotini Coggygiae is to the influence of anoxia in mice time-to-live (x ± s)
Annotate: compare * * P＜0.01, * P＜0.05 with model group.
Ramulus et Folium Cotini Coggygiae has resisting oxygen lack, and when big or middle dosage, its effect is suitable with the 0.0135g/kg Propranolol.
3 discuss
Ramulus et Folium Cotini Coggygiae can be improved mice normal pressure hypoxia-bearing capability, compares with negative control group, can prolong the anoxia mice time-to-live very significantly (P＜0.001), and resisting oxygen lack is arranged.
Claims (10)
1, the new purposes of effective component of Cotinus coggygria, the FOLIUM ET RAMULUS COTINI that it is characterized in that gathering, stem, branch and root are extracted total flavones as improving the body hypoxia-bearing effect and as the medicine of thrombus effect.
2, the new purposes of effective component of Cotinus coggygria according to claim 1 is characterized in that the medicine as prevention and treatment coronary heart disease, angina pectoris and myocardial infarction.
3, the new purposes of effective component of Cotinus coggygria according to claim 1 is characterized in that the medicine as prevention and treatment cerebral thrombosis and thrombotic disease.
4,, it is characterized in that isolating in the described total flavone of effective component of Cotinus coggygria fisetin and Quercetin as medicinal ingredient according to the new purposes of claim 1,2,3 any described effective component of Cotinus coggygria.
5, improve the pharmaceutical preparation of body hypoxia-bearing effect, it is characterized in that containing and isolate fisetin and Quercetin in the total flavone of effective component of Cotinus coggygria as effective ingredient.
6, the pharmaceutical preparation that has the thrombus effect is characterized in that containing and isolates fisetin and Quercetin in the total flavone of effective component of Cotinus coggygria as effective ingredient.
7, the pharmaceutical preparation of raising body hypoxia-bearing according to claim 5 effect, it is characterized in that to isolate fisetin and Quercetin in the total flavone of effective component of Cotinus coggygria, add pharmaceutically acceptable carrier or complementary composition and be prepared from as effective ingredient.
8, the pharmaceutical preparation with thrombus effect according to claim 6 is characterized in that to isolate Quercetin and fisetin in the total flavone of effective component of Cotinus coggygria as effective ingredient, adds pharmaceutically acceptable carrier or complementary composition and is prepared from.
9, the pharmaceutical preparation of raising body hypoxia-bearing according to claim 7 effect is characterized in that the medicine as prevention and treatment coronary heart disease, angina pectoris and myocardial infarction.
10, according to any described pharmaceutical preparation of claim 5-9, it is characterized in that: described pharmaceutical preparation is any in capsule, granule, tablet, powder, injection, oral liquid and the novel form.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102442987A (en) * | 2010-10-09 | 2012-05-09 | 苏州宝泽堂医药科技有限公司 | Fisetin extraction method |
| CN109369589A (en) * | 2018-10-12 | 2019-02-22 | 王刚 | The synchronization extraction process of smoke tree flavonoid glycoside and its application |
| CN115322042A (en) * | 2022-09-14 | 2022-11-11 | 重庆市林业科学研究院 | Cotinus coggygria leaf color regulating agent and using method thereof |
-
2009
- 2009-01-24 CN CNA2009100210460A patent/CN101474220A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102442987A (en) * | 2010-10-09 | 2012-05-09 | 苏州宝泽堂医药科技有限公司 | Fisetin extraction method |
| CN109369589A (en) * | 2018-10-12 | 2019-02-22 | 王刚 | The synchronization extraction process of smoke tree flavonoid glycoside and its application |
| CN115322042A (en) * | 2022-09-14 | 2022-11-11 | 重庆市林业科学研究院 | Cotinus coggygria leaf color regulating agent and using method thereof |
| CN115322042B (en) * | 2022-09-14 | 2023-10-03 | 重庆市林业科学研究院 | Cotinus coggygria leaf color regulating agent and application method thereof |
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