CN101152223A - Use of poplar leaf phenols extract in treating cardiovascular disease and extracting method thereof - Google Patents
Use of poplar leaf phenols extract in treating cardiovascular disease and extracting method thereof Download PDFInfo
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Abstract
The invention provides a new purpose of poplar leaf total phenolic extracts to treat cardiovascular diseases and the extraction method. The invention relates to a purpose poplar leaf total phenolic extracts to treat cardiovascular diseases and the extraction method. The invention provides a new purpose of poplar leaf extracts and a high content effective components extraction method. The poplar leaf total phenolic extracts are used to treat cardiovascular diseases. The extraction method is: 1. decoction; 2. decocted fluid filtration and concentration; 3. column chromatography elution; 4. cooling-sedimentation after pH value acid regulation; 5. Cooling-sedimentation after pH value alkali regulation, and cooling-sedimentation after pH value acid regulation with supernatant solution; 6. pump filtration and filtration and washing with alcohol. The content of the phenolic compounds prepared in the extraction method is higher than 95 percent. The extracts of the invention has the functions of resisting inflammatory, activating blood, removing stagnated blood, antioxidation, alleviating myocardial ischemia, improving the symptoms of arrhythmia and enhancing anoxia test of patients, and is applicable to treat cardiovascular diseases like coronary heart disease, etc.
Description
Technical field
The present invention relates to purposes and the extracting method thereof of a kind of Folium Populi Pseudo-simonii extract in the treatment disease.
Background technology
Along with the raising of expanding economy, living standards of the people and the aggravation of socialization process, the sickness rate of cardiovascular disease rises year by year, has become human main causes of death, and the people's life and health in serious threat.According to the statistics of WHO, because of the number of cardiovascular and cerebrovascular disease death accounts for 28.8% of total death toll to global various diseases death.Expect 2008, this ratio will rise to 36%, become " the No.1 killer " that threaten human health, cause huge spirit and financial burden for the people and society.At present clinically to the treatment of cardiovascular disease based on Western medicine, comprise beta-blocker, calcium ion antagonist, angiotensin converting enzyme inhibitor, angiotensin ii receptor antagonist and HMG-CoA reductase inhibitor etc., but take these medicines and not only easily produce drug resistance, and toxic side effect, even cause patient's arrhythmia.
Though Folium Populi Pseudo-simonii is not record in " Chinese medicine voluminous dictionary ", folk medicine is thought Folium Populi Pseudo-simonii bitter in the mouth, cold in nature, but treating disorders in five ZANG-organs wind heat poison, the detoxifcation relieving constipation.Medical research in recent years finds that the leaf protein of Folium Populi Pseudo-simonii can decompose corrupt protein, and putrefaction removing and tissue regeneration promoting is arranged, and promotes the effect of ulcer wound healing; Also contain sterilization, antipathogenic composition in the Folium Populi Pseudo-simonii, the effect of infection and help muscle and skin growth is arranged.But there is not report up to now about Folium Populi Pseudo-simonii and extract for treating cardiovascular disease thereof; And owing to contained substance classes in the Folium Populi Pseudo-simonii is various, ingredient is extracted and is had the low defective of effective component content.
Summary of the invention
The objective of the invention is to propose a kind of new purposes of Folium Populi Pseudo-simonii extract and solve the low defective of active constituent content in the ingredient extraction, and purposes and the extracting method thereof of a kind of poplar leaf phenols extract that provides in the treatment cardiovascular disease.
Poplar leaf phenols extract of the present invention is used for the treatment of cardiovascular disease.
Above-mentioned poplar leaf phenols extract extracts according to the following steps: one, decoct 2～3 times through Folium Populi Pseudo-simonii water or alcohol after cleaning, pulverizing, decoct 2～4h at every turn, the weight of water or alcohol is 8～14 times of Folium Populi Pseudo-simonii weight; Two, decoction liquor is filtered, then concentrated filtrate proportion to 1.10～1.15; Three, eluting: will be through spissated filtrate through macroporous adsorbent resin column chromatography, water and concentration are 30%～50% ethanol elution successively; Four, concentrate ethanol elution component proportion to 1.15～1.18, add isopyknic distilled water then, reuse hydrochloric acid is regulated pH value to 1～3, is cooled to below 20 ℃ standing sedimentation 12～24h; Five, in the solid phase precipitation thing, add isopyknic deionized water after the abandoning supernatant, and regulating pH value to 6～7 with NaOH solution, standing sedimentation 12～24h extracts supernatant more then, regulate supernatant pH value to 1～3 with hydrochloric acid, afterwards standing sedimentation 12～24h again; Six, sucking filtration obtains precipitate, and with concentration be 95% washing with alcohol precipitate to neutral, promptly obtain poplar leaf phenols extract.
Through chromatography, comprise polyphenol and flavone compound in the poplar leaf phenols extract of the present invention.The content of phenolic compounds that extracting method of the present invention obtains is up to more than 95%.
The effect that poplar leaf phenols extract of the present invention has antiinflammatory, blood circulation promoting and blood stasis dispelling, antioxidation, alleviate myocardial ischemia, improves the arrhythmia symptom and improve patient's hypoxia-bearing capability can be used for treating cardiovascular disease such as coronary heart disease.
The specific embodiment
The specific embodiment one: the present embodiment poplar leaf phenols extract is used for the treatment of cardiovascular disease.
The present embodiment extract adds adjuvant and adopts conventional preparation technology to make different peroral dosage form (oral liquids, tablet, powder, pill or capsule), as add wetting agent (water or water honey) and binding agent (Mel, rice is stuck with paste or batter) make pill, add filler (starch, dextrin, Icing Sugar, lactose, inorganic salts [calcium sulfate, medicinal calcium carbonate], microcrystalline Cellulose or mannitol) and wetting agent (distilled water or ethanol), binding agent (starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methylcellulose, ethyl cellulose, hydroxypropyl emthylcellulose or gelatin solution) etc. make tablet.
The specific embodiment two: the difference of the present embodiment and the specific embodiment one is: poplar leaf phenols extract is used for the treatment of coronary heart disease.
The specific embodiment three: the present embodiment poplar leaf phenols extract extracts according to the following steps: one, decoct 2～3 times through Folium Populi Pseudo-simonii water or alcohol after cleaning, pulverizing, decoct 2～4h at every turn, the weight of water or alcohol is 8～14 times of Folium Populi Pseudo-simonii weight; Two, decoction liquor is filtered, then concentrated filtrate proportion to 1.10～1.15; Three, eluting: will be through spissated filtrate through macroporous adsorbent resin column chromatography, water and concentration are 30%～50% ethanol elution successively; Four, concentrate ethanol elution component proportion to 1.15～1.18, add isopyknic distilled water then, reuse hydrochloric acid is regulated pH value to 1～3, is cooled to below 20 ℃ standing sedimentation 12～24h; Five, in the solid phase precipitation thing, add isopyknic deionized water after the abandoning supernatant, and regulating pH value to 6～7 with NaOH solution, standing sedimentation 12～24h extracts supernatant more then, regulate supernatant pH value to 1～3 with hydrochloric acid, afterwards standing sedimentation 12～24h again; Six, sucking filtration obtains precipitate, and with concentration be 95% washing with alcohol precipitate to neutral, promptly obtain poplar leaf phenols extract.
Present embodiment is measured under 40 ℃ of temperature and is reclaimed cream proportion.After testing in the present embodiment poplar leaf phenols extract phenolic compound purity up to more than 95%.Contain flavone in the present embodiment extract.
The specific embodiment four: the difference of the present embodiment and the specific embodiment three is: alcohol is that concentration is 10%～95% ethanol, methanol or propanol in the step 1.Other step and parameter are identical with embodiment three.
The specific embodiment five: the difference of the present embodiment and the specific embodiment three is: the macroporous resin column in the step 3 is a D101 type macroporous resin column.Other step and parameter are identical with embodiment three.
The specific embodiment six: the difference of the present embodiment and the specific embodiment three is: the eluting concentration of alcohol in the step 3 is 30%.Other step and parameter are identical with embodiment three.
The specific embodiment seven: the difference of the present embodiment and the specific embodiment three is: step 2 concentrated filtrate proportion to 1.14.Other step and parameter are identical with embodiment three.
The specific embodiment eight: the difference of the present embodiment and the specific embodiment three is: step 4 concentrates ethanol elution component proportion to 1.16～1.17.Other step and parameter are identical with embodiment three.
The specific embodiment nine: the difference of the present embodiment and the specific embodiment three is: regulate pH value to 6.8 with NaOH solution in the step 5.Other step and parameter are identical with embodiment three.
The specific embodiment ten: the difference of the present embodiment and the specific embodiment three is: regulate supernatant pH value to 1.6～1.8 with hydrochloric acid in the step 5.Other step and parameter are identical with embodiment three.
The specific embodiment 11: the difference of the present embodiment and the specific embodiment three is: regulate pH value to 1.6～1.8 with hydrochloric acid in the step 4.Other step and parameter are identical with embodiment three.
The specific embodiment 12: the difference of present embodiment and the specific embodiment three or 11 is: the concentration of hydrochloric acid is 3%～5% in the step 4.Other step and parameter are identical with the embodiment hentriaconta-.
The specific embodiment 13: present embodiment and the specific embodiment three or nines' difference is: the concentration of NaOH solution is 4% in the step 5.Other step and parameter are identical with embodiment three or nine.
The specific embodiment 14: present embodiment and the specific embodiment three or tens' difference is: the concentration of hydrochloric acid is 3%～5% in the step 4.Other step and parameter are identical with embodiment three or ten.
The specific embodiment 15: the difference of the present embodiment and the specific embodiment three is: Folium Populi Pseudo-simonii is pulverized the back particle diameter and is not less than 100 orders in the step 1.Other step and parameter are identical with embodiment three.
The specific embodiment 16: the present embodiment poplar leaf phenols extract extracts according to the following steps: one, decoct 3 times with 30% ethanol through the Folium Populi Pseudo-simonii after cleaning, pulverizing, decoct 3h at every turn, alcoholic acid weight is 10 times of Folium Populi Pseudo-simonii weight; Two, decoction liquor is filtered, then concentrated filtrate proportion to 1.12 (measuring under 40 ℃ of temperature); Three, eluting: will be through spissated filtrate through D101 type macroporous adsorbent resin column chromatography, water and concentration are 30% ethanol elution successively; Four, concentrate ethanol elution component proportion to 1.16 (measuring under 40 ℃ of temperature), add isopyknic distilled water then, reuse hydrochloric acid is regulated pH value to 1.5, is cooled to below 20 ℃ standing sedimentation 12h; Five, add isopyknic deionized water after the abandoning supernatant in the solid phase precipitation thing, and regulate pH value to 6.8 with NaOH solution, standing sedimentation 12h extracts supernatant more then, with hydrochloric acid adjusting supernatant pH value to 1.75, and standing sedimentation 12h more afterwards; Six, sucking filtration obtains precipitate, and with concentration be 95% washing with alcohol precipitate to neutral, promptly obtain poplar leaf phenols extract.
Pharmacological testing:
1, hypoxia-bearing capability test
40 healthy mices are divided into 4 groups at random, and (first group is that matched group, second group are that isoproterenol group, the 3rd group are that poplar leaf phenols extract low dose group, the 4th group are 10 every group of poplar leaf phenols extracts (high dose group).
The 3rd group of mice is by every kilogram of lumbar injection poplar leaf phenols extract of body weight 30mg, the 4th group of mice is by every kilogram of lumbar injection poplar leaf phenols extract of body weight 150mg, first group and the second group of equal lumbar injection isometric(al) of mice normal saline, second and third and four groups of mices are all by body weight subcutaneous injection isoproterenol 20mg/kg, with the increase myocardial oxygen consumption behind the 15min.Through behind the 15min mice is put into the 250mL wide mouthed bottle that fills sodica calx again, after sealing, the record mice time-to-live.Test data is as shown in table 1.
Table 1
| Group | Isoproterenol (mg/kg) | Poplar leaf phenols extract (mg/kg) | Time-to-live (min) | Prolong percentage rate |
| First group | - | - | 46.21± 2.41 | 51.41 ** |
| Second group | 20 | - | 30.52± 3.94 | 0 |
| The 3rd group | 20 | 30 | 42.16± 4.28 | 38.14 * |
| The 4th group | 20 | 150 | 47.95± 4.94 | 57.11 ** |
First group has utmost point significant difference (p＜0.01) with second group of mice time-to-live, the 3rd group with second group of mice time-to-live there were significant differences (p＜0.05), there are utmost point significant difference (p＜0.01) the 4th group and second group of mice time-to-live; Test data shows, the subcutaneous injection isoproterenol can obviously shorten the time-to-live of mice under the normobaric hypoxia state, the 3rd group and the 4th group of time-to-live of mice under the normobaric hypoxia state all have obvious antagonism, illustrate that the poplar leaf phenols extract that present embodiment is prepared can obviously improve the mice hypoxia-bearing capability.
2, myocardial preservation experiment
40 healthy Wistar rats are divided into 4 groups at random, and (the 1st group is that matched group, the 2nd group are that pituitrin group, the 3rd group are that poplar leaf phenols extract low dose group, the 4th group are 10 every group of poplar leaf phenols extracts (high dose group).
The 3rd group of rat got thing 20mg by every kilogram of lumbar injection poplar leaf phenols of body weight, the 4th group of rat is by every kilogram of lumbar injection poplar leaf phenols extract of body weight 100mg, the 1st group and the 2nd group of equal lumbar injection isometric(al) of rat normal saline, 20% urethane (5mL/kg) intraperitoneal injection of anesthesia that behind the 1h is with concentration, anesthetized rat is faced upward the position be fixed on the Mus platform and and link to each other, trace electrocardiogram with BL-420 biological function experimental system.The 1st group of rat sublingual vein injecting normal saline, the 2nd, 3 and 4 group of rat sublingual vein injection of pituitrin 1U/kg, be respectively injection at once, observe electrocardiogram behind the injection 1min and change.Change into index with T ripple and ST section, judge degree of myocardial ischemia and drug effect.Abdominal aortic blood behind the injection 60min, centrifugalize serum is measured lactic acid dehydrogenase (LDH), superoxide dismutase (SOD), malonaldehyde (MDA) content in the serum, and test data is as shown in table 2.
Table 2
| Group | LDH(U/L) | MDA(nmol/mL) | SOD(U/mL) |
| The 1st group | 8691.45±1513.32 | 2.69±0.58 | 94.34±9.49 |
| The 2nd group | 10831.37±1301.92 | 5.01±1.45 | 78.35±8.69 |
| The 3rd group | 9189.451703.36 | 3.09±0.79 | 81.15±8.74 |
| The 4th group | 8997.58±1592.64 | 3.08±0.83 | 90.76±9.43 |
The poplar leaf phenols extract that test data explanation present embodiment is prepared can obviously improve the situation of acute myocardial ischemia, the protection cardiac muscle.
30 female Sexual health Wistar rats are divided into 3 groups at random, and (first group is that matched group, second group are that poplar leaf phenols extract low dose group, the 3rd group are 10 every group of poplar leaf phenols extracts (high dose group).Second group of mice is by every kilogram of lumbar injection poplar leaf phenols extract of body weight 20mg, the 3rd group of mice is by every kilogram of lumbar injection poplar leaf phenols extract of body weight 100mg, the first group of equal lumbar injection isometric(al) of rat normal saline, 30min pneumoretroperitoneum injection urethane 1g/kg anaesthetizes rat, FM6501 multi-parameter patient monitoring instrument with the production of development zone, Tianjin Foster development in science and technology company limited is observed the II lead electrocardiogram continuously, import the aconitine normal saline solution by femoral vein with the 1.25ug/min constant speed, write down each Mus arrhythmia time of occurrence respectively and bring out premature ventricular beat (VP), ventricular tachycardia (VT), ventricular fibrillation (VF), the aconitine consumption of asystole (CA), experimental data is as shown in table 3.
Table 3
| Group | Arrhythmia interval (t/min) | Aconitine implantation dosage (ρ B/ μ gkg) | |||
| VP | VT | VF | CA | ||
| First group | 6.04±1.27 | 40.2±6.8 | 49.0±4.1 | 72.1±7.9 | 123.4±9.7 |
| Second group | 7.35±1.34 * | 49.1±5.4 * | 59.8±7.4 * | 90.8±9.7 * | 154.5±19.7 * |
| The 3rd group | 8.01±1.27 * | 50.9±7.3 * | 61.5±6.9 * | 95.0±11.3 * | 183.9±23.8 * |
Second group, the 3rd group with the arrhythmia interval of first group of rat and bring out the aconitine implantation dosage of premature ventricular beat (VP), ventricular tachycardia (VT), ventricular fibrillation (VF), asystole (CA) there were significant differences (p＜0.05); illustrate that the poplar leaf phenols extract that present embodiment is prepared can obviously improve the arrhythmia symptom, have the effect of protection cardiac muscle.
3, antiinflammatory test
40 healthy mices (body weight is 17～23g, male and female half and half) are divided into 4 groups at random, and (first group is that normal saline group, second group of aspirin group, the 3rd group are that poplar leaf phenols extract low dose group, the 4th group are 10 every group of poplar leaf phenols extracts (high dose group).Every day first group of mice normal saline irritate stomach once, second group of aspirin in mouse (200mg/kg) irritate stomach once, the 3rd group of mice poplar leaf phenols extract (20mg/kg) irritate stomach once, the 4th group of mice poplar leaf phenols extract (130mg/kg) irritate stomach once; Gastric infusion is 4 days continuously, the tail vein injection concentration is 0.5% the blue normal saline solution (0.1mL/10g) of ivens behind the last administration 30min, lumbar injection concentration is 0.8% glacial acetic acid physiological salt liquid (0.1mL/10g) immediately then, after 20 minutes mice being taken off cervical vertebra puts to death, cut off skin of abdomen muscle, with 6mL normal saline washing abdominal cavity, collect cleaning mixture with suction pipe, merge the back and add normal saline to 10mL, again with the centrifugal 15min of the rotating speed of 3000r/min, get colorimetric determination trap under the ultraviolet light that supernatant is 590nm in wavelength, experimental data is as shown in table 4.
Table 4
| Group | The OD value |
| First group | 0.35±0.07 |
| Second group | 0.20±0.04 * |
| The 3rd group | 0.23±0.08 * |
| The 4th group | 0.19±0.09 * |
Second and third, four groups with first group of mice OD value there were significant differences (p＜0.05), illustrate that the poplar leaf phenols extract that present embodiment is prepared has antiinflammatory action, can suppress Dichlorodiphenyl Acetate induced mice abdominal cavity capillary permeability and increase.
Claims (10)
1. the purposes of poplar leaf phenols extract in the treatment cardiovascular disease is characterized in that poplar leaf phenols extract is used for the treatment of cardiovascular disease.
2. the purposes of poplar leaf phenols extract according to claim 1 in the treatment cardiovascular disease is characterized in that poplar leaf phenols extract is used for the treatment of coronary heart disease.
3. the extracting method of poplar leaf phenols extract as claimed in claim 1, it is characterized in that poplar leaf phenols extract extracts according to the following steps: one, decoct 2～3 times through Folium Populi Pseudo-simonii water or alcohol after cleaning, pulverizing, each 2～4h that decocts, the weight of water or alcohol is 8～14 times of Folium Populi Pseudo-simonii weight; Two, decoction liquor is filtered, then concentrated filtrate proportion to 1.10～1.15; Three, eluting: will be through spissated filtrate through macroporous adsorbent resin column chromatography, water and concentration are 30%～50% ethanol elution successively; Four, concentrate ethanol elution component proportion to 1.15～1.18, add isopyknic distilled water then, reuse hydrochloric acid is regulated pH value to 1～3, is cooled to below 20 ℃ standing sedimentation 12～24h; Five, in the solid phase precipitation thing, add isopyknic deionized water after the abandoning supernatant, and regulating pH value to 6～7 with NaOH solution, standing sedimentation 12～24h extracts supernatant more then, regulate supernatant pH value to 1～3 with hydrochloric acid, afterwards standing sedimentation 12～24h again; Six, sucking filtration obtains precipitate, and with concentration be 95% washing with alcohol precipitate to neutral, promptly obtain poplar leaf phenols extract.
4. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that alcohol is that concentration is 10%～95% ethanol, methanol or propanol in the step 1.
5. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that the macroporous resin column in the step 3 is a D101 type macroporous resin column.
6. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that step 2 concentrated filtrate proportion to 1.12.
7. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that step 4 concentrates ethanol elution component proportion to 1.16～1.17.
8. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that regulating pH value to 6.8 with NaOH solution in the step 5.
9. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that regulating supernatant pH value to 1.6～1.8 with hydrochloric acid in the step 5.
10. the extracting method of poplar leaf phenols extract according to claim 3 is characterized in that regulating pH value to 1.6～1.8 with hydrochloric acid in the step 4.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104352874A (en) * | 2014-11-26 | 2015-02-18 | 薛学明 | Medicine for treating obesity and preparation method of medicine |
| CN104383334A (en) * | 2014-11-30 | 2015-03-04 | 薛学明 | Medicine for cleaning intestinal toxins and preparation method of medicine |
| CN104623587A (en) * | 2014-12-18 | 2015-05-20 | 青岛市中心医院 | Chinese medicinal preparation for treating cardiovascular diseases and preparation method of Chinese medicinal preparation |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1069535C (en) * | 1998-11-20 | 2001-08-15 | 阎凤海 | Clinical medicinal prepns. contg. flavongly coside extracted for persimmon leaves, and process for preparing same |
| CN1093409C (en) * | 1999-08-16 | 2002-10-30 | 王在田 | Fracture treating plaster |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104352874A (en) * | 2014-11-26 | 2015-02-18 | 薛学明 | Medicine for treating obesity and preparation method of medicine |
| CN104383334A (en) * | 2014-11-30 | 2015-03-04 | 薛学明 | Medicine for cleaning intestinal toxins and preparation method of medicine |
| CN104623587A (en) * | 2014-12-18 | 2015-05-20 | 青岛市中心医院 | Chinese medicinal preparation for treating cardiovascular diseases and preparation method of Chinese medicinal preparation |
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