CN1290518C - Extracting method of propolis effective component and its use - Google Patents
Extracting method of propolis effective component and its use Download PDFInfo
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- CN1290518C CN1290518C CNB2004100258363A CN200410025836A CN1290518C CN 1290518 C CN1290518 C CN 1290518C CN B2004100258363 A CNB2004100258363 A CN B2004100258363A CN 200410025836 A CN200410025836 A CN 200410025836A CN 1290518 C CN1290518 C CN 1290518C
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- propolis
- extract
- present
- water
- potassium chloride
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Abstract
The present invention relates to an extraction method of effective components of bee glue and the application of the effective components. More specifically, the present invention relates to a method for extracting water soluble components in the bee glue and the application of the water soluble components of the bee glue. The extraction method is characterized in that the bee glue is extracted by water. The water soluble extract products obtained by the extraction method can be applied to the following two aspects: the preparation of medicine for treating coronary artery relaxation and the preparation of phytoestrogen medicine.
Description
Technical field
The present invention relates to the extracting method of active ingredient in a kind of propolis, and the purposes of this active ingredient.More exactly, the present invention relates to extract the purposes of water-soluble components in the method for water-soluble components in the propolis and the propolis that proposed.
Background technology
Propolis is the resin secretions and the Cera Flava of Apis herborization, through chewing a kind of resinoid that modulation forms.Composition is very complicated in the propolis, and both at home and abroad to the chemical composition of propolis, biological agent, pharmacological action have a lot of research reports in recent years.It is antibiotic to discover that propolis has, antiviral, antiulcer, antiinflammatory, local anesthesia, blood pressure lowering, effects such as enhancing immunity.For active ingredient in the propolis, concentrate on organic solutions such as ethanol and ether in the prior art and extract, perhaps extract with high concentration organic solution soluble in water.As disclosed content in the Chinese invention patent 97111643.1.This patent adopts acetone, or acetic acid, or ethanol, or methanol, or isopropyl alcohol, perhaps with the active ingredient 3-[4-3 in the mixture extraction propolis of aforementioned solvents, and 5 hydroxyls-(two methyl-2 cyclobutenyls of 3-) phenyl]-2-acrylic acid, be used for anticorrosion, hair growth promoting and anticancer.In the prior art propolis extracted with non-organic solution and do not appear in the newspapers.
Summary of the invention
First purpose of the present invention provides a kind of water-soluble components method of extracting from propolis; Second purpose of the present invention is the purposes of the water-soluble components extracted from propolis of open usefulness method of the present invention.
Method of the present invention is that the propolis water is extracted.
The present invention more specifically method for optimizing is that propolis is pulverized, and water extracts again, and extracting solution is got its supernatant through centrifugal or abundant post precipitation, again with the supernatant lyophilizing.
Purposes of the present invention is meant with application and the application in preparation plant estrogens medicine of water solubility extract in the resulting propolis of method of the present invention in preparation coronary artery diastole medicine.
Extracting method of the present invention is very simple, and extraction cost is low.Propolis extract after the inventive method is handled is formed simple.Can also continue after adopting method of the present invention to handle therefore can make full use of the propolis resource with other active ingredient in the prior art extraction propolis.In addition, extract of the present invention can be used for preparing coronary artery diastole medicine, does not appear in the newspapers in this point prior art.Extract of the present invention is used for preparing the application of plant estrogens medicine, for the production of estrogens medicine provides new approach and raw material sources.
Description of drawings
Fig. 1 is the thin slice scan result of Flavonoid substances in the propolis extract.
Fig. 2 is that propolis extract concentration of the present invention is to direct diastole effect curves coronarius.
Fig. 3 is that propolis extract concentration of the present invention is to the pre-shrunk coronary artery inhibitory action of potassium chloride curve.
Fig. 4 is a nitric oxide synthase inhibitor activity, dispels coronary artery endothelial, methylene blue, and propranolol and indomethacin are to the direct diastole function influence of propolis extract of the present invention curve.
Fig. 5 is a nitric oxide synthase inhibitor activity, dispels coronary artery endothelial, methylene blue, propranolol and indomethacin to propolis extract of the present invention to the pre-shrunk inhibiting influence curve coronarius of potassium chloride.
Fig. 6 is the influence of propolis extract of the present invention to porcine coronary potassium chloride shrinkage curve.
Fig. 7 is the influence of propolis extract of the present invention to porcine coronary calcium chloride shrinkage curve.
The specific embodiment
The result of the test of following detailed explanation method of the present invention and associated uses.
One, propolis extracting method
Get fresh propolis, pulverize and be placed on distilled water (W/W=1: 20) under room temperature, stirred 48 hours; Filter; Filtering residue adds water and extracts repeatedly, filters; Merging filtrate, 15, under the 000rpm condition centrifugal 60 minutes;
Merge supernatant; After handling, lyophilizing gets the water-soluble bee glue composition.
After measured, the water-soluble bee glue that is extracted contains three kinds of flavone components, referring to accompanying drawing 1.Fig. 1 is the result of flavone thin slice scan under visible wavelength in the propolis water extract, and used developer is ferric chloride-alcoholic solution of 5%.
Two, propolis extract purposes of the present invention research
Experimental technique and condition:
Get the Cor Sus domestica of just having slaughtered from pig slaughtering factory and exist side by side soon that Cor Sus domestica places 0 ℃ normal saline, be transported to laboratory then immediately.At room temperature the pig left anterior descending coronary artery is cut and places the Krebs liquid of the carbon dioxide that is connected with 95% oxygen and 5% from cardiac muscle, temperature remains on 37 ℃.Carefully wipe out fat and its hetero-organization thereof of being attached on the arteria coronaria.Being cut into length then is the long coronary artery ring of 4-5mm.Immediately arterial ring is placed 10ml glass bath pipe with two parallel stainless hook levels, bathe in the pipe and contain Krebs liquid, it consists of (mM) NaCl 118, and KCl 4.7, NaH
2PO
41.2, MgCl
21.2, NaHCO
315.4, CaCl
22.5 and glucose 11.5, and continue to feed 95%O
2And 5%CO
2Mist, temperature remains on 37 ℃ in whole experiment.Preload gives 2g.Blood vessel produces tension force and is recorded in computer by pressure receptor (JH-2) with BL-410E biotic experiment system.
1, the propolis water extract is to direct effect coronarius
The porcine coronary ring hatched 90 minutes in bathing pipe after, again the propolis water extract is added the balneation pipe successively by the 0.316-100ug/ml variable concentrations, normal saline with equivalent adds another group porcine coronary ring at the bath pipe of another similarity condition simultaneously, as blank contrast experiment.Observation experiment result, result show that the propolis water extract can dose-dependent diastole coronary artery EC50=(75.35 ± 1.24) ug/ml (n=8).Show directly diastole coronary artery of propolis water extract, its maximum diastolic rate reaches 56.33%, referring to accompanying drawing 2.NS is the normal saline group among the figure, and WEP is a propolis water extract group.
2, the propolis water extract is to the pre-shrunk effect coronarius of potassium chloride
The porcine coronary ring hatched 90 minutes in bathing pipe after, add KCl (40mM) preshrinking coronary artery, after 15 minutes the propolis water extract is added the balneation pipe successively by the variable concentrations of 0.316-100ug/ml, normal saline with equivalent adds another group porcine coronary ring at the bath pipe of another similarity condition simultaneously, as blank contrast experiment.Observation experiment result, result show that the propolis water extract can dose-dependent diastole coronary artery IC50=(65.11 ± 1.72) ug/ml (n=8).Show that propolis can effectively suppress the contraction that causes because of potassium chloride, its maximum diastolic rate reaches 64.77%, referring to accompanying drawing 3.NS is the normal saline group among the figure, and WEP is a propolis water extract group.
3, nitric oxide synthase inhibitor activity is dispeled coronary artery endothelial, methylene blue, and propranolol and indomethacin are to the influence of propolis water extract diastole effect
Prepare coronary artery and before being cut to vascular ring, wipe blood vessel endothelium gently, to measure of the diastole effect of propolis water extract it with cotton swab.
The coronary blood pipe ring of not dispeling coronary artery endothelial is tested its function with Kallidin I, think that after tested the good vascular ring of endothelial function is as carrying out the experiment to the influence of propolis water extract diastole effect of nitric oxide synthase inhibitor activity, methylene blue, propranolol and indomethacin.
With nitric oxide synthase inhibitor activity (10
-4M), methylene blue (10
-5M), propranolol (10
-5M) and indomethacin (10
-5M) adding propolis water extract (100ug/ml) 15 minutes before respectively, perhaps add and added the balneation pipe before in 15 minutes at potassium chloride (40mM), the experimental group that adds potassium chloride also adds the balneation pipe with propolis water extract (100ug/ml) again adding potassium chloride after 15 minutes.Test result shows, nitric oxide synthase inhibitor activity, dispel endothelium and methylene blue and all can significantly lower the propolis water extract, or the propolis water extract is had obvious suppression effect (all P values are all less than 0.05) to the diastole effect of the pre-shrunk arteria coronaria of potassium chloride direct diastole effect coronarius; Propranolol and indomethacin then do not have this effect (all P values are all greater than 0.05), referring to accompanying drawing 4 and Fig. 5.
In accompanying drawing 5 and Fig. 6: L-NNA is the nitric oxide synthase inhibitor activity experimental group; MB is the methylene blue experimental group; Prop is the propranolol experimental group; Indo is the indomethacin experimental group; Denu is for dispeling the coronary artery endothelial experimental group.
4, the propolis water extract is to the inhibitory action of potassium chloride concentration dependency shrinkage curve
Successively the concentration of potassium chloride by 4-100mM is added the coronary blood pipe ring hatched 90 minutes in bathing pipe after, obtain potassium chloride dose dependent shrinkage curve, fully flushing returns to original state down to tension force then, then propolis water extract (100ug/ml) is added the balneation pipe, again potassium chloride is added successively by 4-100mM concentration again after 20 minutes.For comparing with experimental result, other establishes the blank experimental group, promptly adds another with the equivalent normal saline under condition of equivalent and organizes crown blood arterial ring.The result shows that the propolis water extract can significantly suppress the inhibition curve of potassium chloride.The EC50 value of potassium chloride curve is that (n=8, control), the EC50 value of the potassium chloride curve after propolis water extract (100ug/ml) suppresses is that (n=8, p＜0.05vs.control) is referring to accompanying drawing 6 for (93.12 ± 1.21) mM to (16.07 ± 1.42) mM.
In the accompanying drawing 7: KCl is the potassium chloride shrinkage curve; KCl+WEP is for successively adding the shrinkage curve of propolis and potassium chloride; NS is the curve of blank.
6, the propolis water extract is to the inhibitory action of calcium chloride concentration dependency shrinkage curve
The coronary artery ring places no calcium Krebs liquid, wherein contains the EGTA of 0.1mM, adds potassium chloride (80mM) after hatching 90 minutes, adds by 10 successively after 15 minutes
-5-10
-2The calcium chloride of M obtains calcium chloride dose dependent shrinkage curve, and then fully flushing is tried the coronary vasodilator ring, so that its tension force returns to original state, again propolis water extract (100ug/ml) is added the balneation pipe, again calcium chloride is pressed 10 after 20 minutes
-5～10
-2The concentration of M adds again successively.For comparing with experimental result, for comparing with experimental result, other establishes the blank experimental group, promptly adds another with the equivalent normal saline under condition of equivalent and organizes crown blood arterial ring.Experimental result shows that the propolis water extract can significantly suppress the inhibition curve of calcium chloride.The EC50 value that calcium chloride suppresses curve is (3.76 ± 0.58) * 10
-3(n=8, control), the EC50 value of the potassium chloride curve after propolis water extract (100ug/ml) suppresses is (7.98 ± 1.24) * 10 to M
-3M (n=8, p＜0.05vs.control), ginseng accompanying drawing 7.
In the accompanying drawing 7: Calcium is the calcium chloride shrinkage curve; Calcium+WEP is for successively adding the shrinkage curve of propolis and potassium chloride; NS is the curve of blank.
Because the propolis extract of the present invention diastole effect good to coronary artery, therefore propolis extract of the present invention has manifested the good prospect of the medicine that can be used as treatment coronary heart disease.So the application prospect of propolis water extract is widely, be huge particularly to its value on the therapeutic use of coronary heart disease.Women's cause of coronary heart disease after menopause significantly rises in addition, and estrogenic alternative medicine can obviously reduce its sickness rate, but women's breast tumor sickness rate is improved.The phytoestrogen of available technology adopting but can be avoided this harmful effect.The propolis water-soluble flavone class material that the present invention extracts from propolis extract, this type of material is the material with estrogen-like effects equally, so the present invention provides new production approach and the new raw material of a class for the preparation of plant estrogens medicine.
Claims (1)
1. the application of the extract that extracts of propolis water in preparation coronary artery diastole medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100258363A CN1290518C (en) | 2004-01-08 | 2004-01-08 | Extracting method of propolis effective component and its use |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100258363A CN1290518C (en) | 2004-01-08 | 2004-01-08 | Extracting method of propolis effective component and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1555814A CN1555814A (en) | 2004-12-22 |
| CN1290518C true CN1290518C (en) | 2006-12-20 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100258363A Expired - Fee Related CN1290518C (en) | 2004-01-08 | 2004-01-08 | Extracting method of propolis effective component and its use |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102005031361A1 (en) * | 2005-06-30 | 2007-01-04 | Biotronik Vi Patent Ag | Use of propolis as a coating material for medical implants |
| CN102078337A (en) * | 2011-01-14 | 2011-06-01 | 哈药集团密山蜂业有限公司 | Propolis natural reduction extraction process method |
| CN103125802B (en) * | 2013-03-11 | 2014-07-30 | 宁夏原野蜂业有限公司 | Nutrition taking propolis cream, medlar cream, honey and desert ginseng powder as raw materials and preparation method of nutrition |
| CN104557520B (en) * | 2015-01-21 | 2016-01-13 | 聊城大学 | A kind of method of separation and purification coffic acid, forulic acid and isoferulic acid from Chinese Water extracts of propolis |
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2004
- 2004-01-08 CN CNB2004100258363A patent/CN1290518C/en not_active Expired - Fee Related
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| CN1555814A (en) | 2004-12-22 |
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