CN101450092A - Vladimiria root extract, composition containing the vladimiria root extract and use thereof - Google Patents
Vladimiria root extract, composition containing the vladimiria root extract and use thereof Download PDFInfo
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Abstract
The invention relates to a vladimiria root extract belonging to medicament field, especially to an extract from vladimiria root used to treat ulcer diseases. The extract using medicinal vladimiria root as material is prepared by: extracting by supercritical fluid or alcohols solvent, suspending by water, extracting by nonpolar organic solvent and separating, collecting the organic phase part and drying. The extracting method in the invention can achieves vladimiria root extract with stable controllable quality, can make the cost largely lower than the single compound preparation. The exact prepared by the process of the invention achieves evident curative effect for treating ulcer diseases, and provides another selection for treating ulcer diseases in clinic.
Description
Technical field
The present invention relates to a kind of Radix Vladimiriae extract, specifically, is that Radix Vladimiriae is that the raw material extraction is prepared from, and can be used for preparing the treatment for the treatment of Peptic Ulcers, belongs to field of medicaments.
Background technology
The peptic ulcer major part occurs in stomach or duodenum, is called for short Peptic Ulcers, and clinical manifestation is based on stomachache, and pain is limited to epigastrium, has chronic, periodically and 3 characteristics of rhythmicity.Peptic Ulcers is the commonly encountered diseases of internal medicine.Data shows: sickness rate is up to 20%, and it is longer to show as the course of disease, and is changeable repeatedly, and complication sees more, has a strong impact on people's work and quality of life.The active drug of Chinese traditional treatment Peptic Ulcers such as CHENXIANGLU BAILU PIAN; the upright health sheet of stomach etc.; Western medicine such as ranitidine capsule; losec capsule etc.; statistics shows in the chemicals commonly used (except the mucosa protective agent); mechanism of action difference but exercising result is all identical; by changing the acid environment of stomach; help ulcer healing; but in therapeutical effect, exist the digestive system untoward reaction without exception; specifically comprise nauseating; vomiting; diarrhoea or constipation etc.; and relapse rate is high, and (with safe stomach U.S.A is example, is generally for 4～6 weeks the course of treatment, and total effective rate is more than 80%; 6 months relapse rates are that 24%, 1 year relapse rate can be up to 85% after the drug withdrawal).[Liu Yutao, Sun Hua. the mechanism of action of treatment peptic ulcer medicine and the relation of its untoward reaction. professional and healthy .2004,20 (8): 128].
Radix Vladimiriae is the dry root of feverfew Radix Vladimiriae Vladimiria souliei (Franch.) Ling and gray wool Radix Vladimiriae Vladimiriasouliei (Franch.) Ling var.cinerea Ling, the function promoting the circulation of QI to relieve pain, being used for the treatment of diseases such as abdominal distention, liver and gall pain, is the medicine commonly used of treatment digestive tract disease.Radix Vladimiriae be in, hide to intersect medicine variety, the clear Baconic's heat of Radix Vladimiriae function all on the books in the classical masterpiece Four-Volume Medical Code of Tibetan medicine, " brilliant pearl book on Chinese herbal medicine ", granulation promoting fat, with control the abdomen flatulence completely, inappetence, gastric ulcer etc.
From Radix Vladimiriae, separate the total lactone position that obtains, form by multiple lactones such as dehydrocostuslactone and costunolide and Radix Vladimiriae lactones, wherein key component is dehydrocostuslactone [Li Zhaolin, Xue Dunyuan, Wang Mingkui, research Lanzhou University journal (natural science edition) 1991 Deng the Radix Vladimiriae chemical composition of volatile oil, 7 (4): 94] [China's book on Chinese herbal medicine (the 7th the 21st volume) Shanghai is compiled by editorial board of State Administration of Traditional Chinese Medicine's " China's book on Chinese herbal medicine ": the 1999:816 of Shanghai science tech publishing house] [Chang Xinquan, the Ding Lixia chief editor. active ingredient of Chinese herbs is analyzed handbook. Beijing: Xueyuan Press, 2002.262].
Adopt the patent application of the derivant of dehydrocostuslactone and costunolide, that directly related is JP05331062 and JP07138157, adopt dehydrocostuslactone as active matter, be used for antiulcer report, as: " Sesquiterpenecompound and antiulcer agent comprising the same as active ingredient (lactone compound and as the purposes of antiulcer medicine active component) ".Japanese patent application publication No.: 06-211826 1994, discloses the new purposes that is used for antiulcer (Antiulcer) with the derivant of dehydrocostuslactone and costunolide; " Antiulcer agent (antiulcer medicine) " (Japanese patent application publication No.: 05331062,1993 year) discloses with the reactive compound of dehydrocostuslactone as the antiulcer medicine; " Anti-ulcer agent (antiulcer medicine) " (Japanese patent application publication No.: 07-138157, nineteen ninety-five), the method for preparing the antiulcer medicine is disclosed, be used for the antiulcer treatment with the complex of dehydrocostuslactone and Polyethylene Glycol.
At present, the existing report that adopts Muxiang total lactone extract to be used for treatment of diseases such as cholecystitis, cholelithiasis, as: " extract of a kind of Radix Aucklandiae total lactones and its production and application " (Chinese patent, application number: 200410014854.1), the total lactone that extracts from the Chinese medicine Radix Aucklandiae is wherein disclosed, wherein contain compositions such as dehydrocostuslactone and costunolide, be used for the treatment of or prevent cholecystitis, cholelithiasis.But the raw material Radix Aucklandiae of this patent disclosure is the dry root of feverfew Aucklandia lappaDecne., the raw material Radix Vladimiriae of adopting with the present invention belongs to equal not generic plant, different on both ingredients and the component content, even both adopt identical or close method for extraction and purification, but the composition in extract obtained also is different with component content, its medicinal effects also has very big difference with extract of the present invention, as the present invention is the treatment that extract is applied to Peptic Ulcers, and this patent is to use total lactone to be used for the treatment of or to prevent cholecystitis, cholelithiasis.
Above-mentioned document and patent documentation have been reported the dehydrocostuslactone in the Radix Vladimiriae, are useful on the pharmacological action that suppresses or suppress experimental gastric ulcer indirectly; The dehydrocostuslactone in the Radix Vladimiriae and the derivant of costunolide are used for the treatment of the purposes of ulcer or Peptic Ulcers; But to from Radix Vladimiriae, extract the monomer of dehydrocostuslactone and costunolide, or and then to prepare the cost of its derivant very high, but the dehydrocostuslactone and the content determination of costunolide that are extracted by Radix Vladimiriae are extremely low.And do not have synthetic method to prepare the relevant report of dehydrocostuslactone and costunolide at present yet.In view of the Chinese crude drug complicated component, the composition of medicine performance drug effect is indeterminate, only finds wherein single component or position, can not accurately judge the drug effect of medicine.
Summary of the invention
It is low that technical problem solved by the invention provides a kind of preparation cost, and can effectively treat the Radix Vladimiriae extract of Peptic Ulcers.
This Radix Vladimiriae extract is to be raw material with the medical material Radix Vladimiriae, and supercritical extraction or add after alcohols solvent extracts adds aqueous suspension, adopts the non-polar organic solvent extract and separate, collects organic facies and partly is drying to obtain; Wherein, described alcohols solvent is ethanol, methanol, isopropyl alcohol etc., because methanol has toxicity, is the extraction solvent from extraction cost and extraction effect aspect preferred alcohol; Described non-polar organic solvent is ether, petroleum ether or ethyl acetate.Medical material Radix Vladimiriae source is the dry root of feverfew Radix Vladimiriae Vladimiria souliei (Franch.) Ling and gray wool Radix Vladimiriae Vladimiria souliei (Franch.) Lingvar.cinerea Ling.
Radix Vladimiriae extract of the present invention is by following method preparation:
A, take by weighing Radix Vladimiriae and pulverize, adopt supercritical extraction method, solvent supersonic extraction method, solvent percolation extraction method or solvent refluxing extraction method to extract, solvent is an alcohols solvent;
B, with the solvent extraction liquid of a step preparation, behind the reclaim under reduced pressure alcohols solvent, add aqueous suspension, adopt the non-polar organic solvent extract and separate, behind the reclaim under reduced pressure non-polar organic solvent, drying under reduced pressure organic facies part promptly gets Radix Vladimiriae extract of the present invention again
According to pharmacodynamics test; Radix Vladimiriae extract of the present invention has tangible promoting the circulation of QI to relieve pain effect; also has tangible antiulcer action; can reach the effect for the treatment of both the principal and secondary aspects of a disease to the treatment of Peptic Ulcers; according to traditional medication custom and early-stage Study; the antiulcer action that compares ranitidine; the Radix Vladimiriae ethyl acetate extract is better than acetic acid type to the effect of reserpine type gastric ulcer; may be to the protective effect of reserpine type ulcer with to improve mechanism of action such as gastrointestinal motility and stomachial secretion relevant, being suitable for the abdominal distention gastric abscess is the Peptic Ulcers of cardinal symptom.From the clinical practice performance analysis, adopt Radix Vladimiriae and preparation determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs on the basis of traditional medication experience thereof, in case of necessity as the adjuvant drug of chemotherapy, avoid long-term but sour treatment back drug withdrawal to cause the rising of gastric acid secretion bounce-back property, reduce the recurrence of ulcer disease rate, reduce untoward reaction, increase the effectiveness of clinical practice.
Radix Vladimiriae extract of the present invention provides a kind of new selection for the clinical treatment Peptic Ulcers, this Radix Vladimiriae method for preparing extractive is simple, Radix Vladimiriae extract stable and controllable for quality of preparation gained, cost is significantly less than preparation unification compound, the cost of preparation Radix Vladimiriae extract is about 0.2 ten thousand yuan/kg, can not be higher than 10,000 yuan/kg (conversion is dehydrocostuslactone) by lower bound 25% accounting price of contained dehydrocostuslactone; The price of dehydrocostuslactone (purity is more than 98%) is 148 a yuan/20mg (national standard product price, price is lower), the monomeric price of conversion back dehydrocostuslactone is 7,400,000 yuan/kg, (with dehydrocostuslactone content in the Radix Vladimiriae extract is basic calculation, the dehydrocostuslactone monomer than the height of Radix Vladimiriae extract price 740 times), the two compares, and the Costco Wholesale of Radix Vladimiriae extract is with the obvious advantage.Radix Vladimiriae extract of the present invention is that the pharmacy quality that is prepared from of active component is controlled, taking convenience, and the medicine bioavailability improves, and drug effect is remarkable.
Description of drawings
Fig. 1 dehydrocostuslactone reference substance chromatogram
Fig. 2 Radix Vladimiriae extract chromatogram
Fig. 3 Radix Vladimiriae extract and effective ingredient cause the inhibitory action of the experimental gastric ulcer of chmice acute to reserpine
Fig. 4 Radix Vladimiriae extract Dichlorodiphenyl Acetate causes the inhibitory action of the experimental gastric ulcer of rat chronic
The specific embodiment
The invention provides a kind of new Radix Vladimiriae extract, it is to be raw material with the medical material Radix Vladimiriae, and supercritical extraction or add after alcohols solvent extracts adds aqueous suspension, adopts the non-polar organic solvent extract and separate, collects organic facies and partly is drying to obtain; Wherein, described alcohols solvent is ethanol, methanol, isopropyl alcohol etc., because methanol has toxicity, is the extraction solvent from extraction cost and extraction effect aspect preferred alcohol; Described non-polar organic solvent is ether, petroleum ether or ethyl acetate.
Concentration of alcohol is 30%～95%, and it is good selecting concentration of alcohol 70～95% according to the total ester amount of index components content.Show that according to comprehensive pharmacological effect experiment ethyl acetate is better as the sick curative effect of comprehensive antiulcer of separating the Radix Vladimiriae extract that gets with the organic solvent extraction preparation.
When the concentration of extracting etoh solvent is 30%～95%, can realizes extracting and contain total ester amount with dehydrocostuslactone C
15H
18O
2Meter is greater than the extract of 50～80%w/w.
Contain total ester amount in the extract with dehydrocostuslactone C
15H
18O
2Count 50～80%w/w, contain dehydrocostuslactone C
15H
18O
2Be 25～70%w/w, total ester amount and dehydrocostuslactone are as the quality control standard of Radix Vladimiriae extract of the present invention; Therefore contained single component amount is not carried out concrete dose-effect relationship relatively as yet in the Radix Vladimiriae extract, and this content is mainly as quality evaluation index.From the data of test the example 3 as can be seen: the result of three administration groups of dehydrocostuslactone monomeric compound does not demonstrate clear and definite dose-effect relationship, in, the effect of low dosage is better relatively, there was no significant difference on the statistics.
Radix Vladimiriae extract of the present invention is by following method preparation:
A, take by weighing Radix Vladimiriae and pulverize, adopt supercritical extraction method, solvent supersonic extraction method, solvent percolation extraction method or solvent refluxing extraction method to extract, solvent is an alcohols solvent;
B, with the solvent extraction liquid of a step preparation, behind the reclaim under reduced pressure alcohols solvent, add aqueous suspension, adopt the non-polar organic solvent extract and separate, behind the reclaim under reduced pressure non-polar organic solvent, drying under reduced pressure organic facies part promptly gets Radix Vladimiriae extract of the present invention again.
Show that according to pharmacological effect experiment the purposes of Radix Vladimiriae extract of the present invention in the medicine of preparation treatment Peptic Ulcers is purposes in the medicine of Peptic Ulcers of cardinal symptom with the abdominal distention gastric abscess in the preparation treatment especially.
The present invention also provides a kind of pharmaceutical composition, it is that Radix Vladimiriae extract with effective dose is an active component, add the medicament that acceptable accessories or complementary composition are prepared from, take dosage forms such as to be prepared into capsule commonly used, soft capsule, tablet, granule, powder, drop pill, oral liquid or pill for the ease of the patient.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.
The specific embodiment of form and screening experiment are described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
One, the extraction process screening test of Radix Vladimiriae extract of the present invention:
Embodiment 1: the preparation of Radix Vladimiriae extract of the present invention
A, take by weighing Radix Vladimiriae and pulverize, adopt the supercritical extraction method to extract; Or
Take by weighing Radix Vladimiriae and pulverize, adopt the solvent supersonic extraction method to extract, used concentration of alcohol is 30%～95%; Or
Take by weighing Radix Vladimiriae and pulverize, adopt solvent percolation extraction method to extract, used concentration of alcohol is 30%～95%; Or
Take by weighing Radix Vladimiriae and pulverize, adopt the solvent refluxing extraction method to extract, used concentration of alcohol is 30%～95%;
B, with the solvent extraction liquid of A step preparation, decompression and solvent recovery adds aqueous suspension, organic solvent extraction separates, drying under reduced pressure behind the reclaim under reduced pressure organic solvent promptly gets Radix Vladimiriae extract of the present invention.
Wherein the supercritical extraction method of Cai Yonging, solvent supersonic extraction method, solvent percolation extraction method or solvent refluxing extraction method are conventional publicly-owned technology.
1, concentration of alcohol screening
Adopt evenly table of design
Arrange optimization Test, test, get Radix Vladimiriae coarse powder 50g, add the concentration of regulation under each experiment numbers item and the ethanol of consumption, load onto reflux condensing tube, reflux condensation mode in water-bath by drafting method in the experiment table.
The total lactone of table 1 Radix Vladimiriae extracts the test uniform designs table
| Numbering | Concentration of alcohol (%) | Consumption (doubly) | Time (h) | Number of |
| 1 | 30 | 4 | 1 | 1 |
| 2 | 30 | 6 | 2 | 1 |
| 3 | 50 | 8 | 1 | 2 |
| 4 | 50 | 10 | 2 | 2 |
| 5 | 70 | 4 | 0.5 | 3 |
| 6 | 70 | 6 | 1.5 | 3 |
| 7 | 90 | 8 | 0.5 | 4 |
| 8 | 90 | 10 | 1.5 | 4 |
Table 2 sample contains ester quantitative determination result
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| Contain ester amount (%) | 4.41 | 9.73 | 23.04 | 15.54 | 11.78 | 19.71 | 20.63 | 27.17 |
Above-mentioned experimental result explanation is when concentration of alcohol is 70～90%, and the ester amount that contains of extract obviously improves, and concentration of alcohol is 90% o'clock, and it is the highest that extract contains the ester amount.
2, supercritical extraction method, solvent supersonic extraction method, three kinds of extracting method yields of solvent percolation extraction method are relatively seen Table 3, wherein supercritical fluid is SF-CO
2, wherein solvent species is 90% ethanol, wherein the sample to solvent supersonic extraction method, solvent percolation extraction method continues to have adopted the ethyl acetate extraction separation.
Table 3 supercritical extraction method, solvent supersonic extraction method, solvent percolation extraction method yield are relatively
| Extracting method | The supercritical extraction method | The solvent supersonic extraction method | Solvent percolation extraction method |
| Extract yield (%) | 5.96 | 6.25 | 5.62 |
| Contain ester amount (%) | 66.11 | 63.20 | 66.90 |
| Dehydrocostuslactone content (%) | 43.27 | 40.60 | 41.48 |
The yield that last table shows three kinds of extracting method is approaching, taking cost into account selects suitable extracting method so can reach according to the simplification of practical operation.
According to the statistical software stepwise regression analysis showed, determine following optimization Test scheme:
Scheme 1: get the Radix Vladimiriae coarse powder, add 90% alcohol reflux secondary, alcohol adding amount is 10 times of amounts, each 1 hour, filter, and get filtrate for later use;
Scheme 2: the extraction of getting scheme 1 remainder filters residue, adds 90% alcohol reflux secondary, and alcohol adding amount is 10 times of amounts, and each 1 hour, filter, get filtrate for later use.
Demonstration test result
The total lactone extraction ratio of the Radix Vladimiriae of scheme 1 measurement result is: 18.38%;
The total lactone extraction ratio of the Radix Vladimiriae of scheme 2 measurement result is: 4.14%.
Total lactone of the present invention is meant the complex that is grouped into by one-tenth such as dehydrocostuslactone, and the described ester amount that contains is meant that this complex accounts for the ratio content of extract.
Draw by above-mentioned data, the content of the total lactone of Radix Vladimiriae that extracts once more in the residue after extracting for the 2nd time seldom, so optimizing technology is scheme 1.
Conclusion (of pressure testing)
The total lactone composition of Radix Vladimiriae is 10 times of amounts with 90% alcohol reflux secondary, alcohol adding amount, and the total lactone extraction ratio of this method Radix Vladimiriae was the highest in each 1 hour, was optimum process; With above gained ethanol extract, decompression recycling ethanol adds aqueous suspension, ethyl acetate extraction, and the reclaim under reduced pressure ethyl acetate, drying under reduced pressure is collected the Radix Vladimiriae extract.
Two, Radix Vladimiriae extract quality control of the present invention
1, total lactone content is measured in the Radix Vladimiriae extract of the present invention
Total ester amount: get embodiment 1 respectively and adopt 90% ethanol refluxing process to prepare the about 0.5g of Radix Vladimiriae extract of gained, accurate claim surely, add ethanol 50ml, put that tepor makes dissolving in the water-bath, put cold, add 10% Lead monosubacetate test solution 5ml, constantly jolting was placed 1 hour, precipitation fully, filter, precipitation ethanol thorough washing merges washing liquid and filtrate, add 25% metabisulfite solution 5ml, placed 2 hours after the jolting, add active carbon 1g again, put and boil 10 minutes in the water-bath, sucking filtration while hot, through being covered with the buchner funnel of 1g active carbon layer, and, merge washing and filtrate with hot ethanol thorough washing active carbon layer, reclaim ethanol to about 50ml, put coldly, add 3 of water 80ml and instructions phenolphthalein solutions, (0.1mol/L) is neutralized to blush with caustic lye of soda.Accurate sodium hydroxide volumetric solution (0.1mol/L) 25ml that adds, the close plug of lucifuge was placed 24 hours, disappeared to redness with hydrochloric acid volumetric solution (0.1mol/L) titration and took off, and titration results is proofreaied and correct with blank assay, promptly.Every 1ml sodium hydroxide volumetric solution (0.1mol/L) is equivalent to 23.1mg dehydrocostuslactone (C
15H
18O
2).
The Radix Vladimiriae extract is pressed dry product and is calculated, and contains the ester amount with dehydrocostuslactone (C
15H
18O
2) calculate, should be not less than 50%.
2, dehydrocostuslactone assay in the Radix Vladimiriae extract of the present invention
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D).
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-water (70:30) is a mobile phase; The detection wavelength is 220nm.Theoretical cam curve is calculated by the dehydrocostuslactone peak should be not less than 6000.
The preparation of reference substance solution: it is an amount of to remove hydrogen constuslactone reference substance, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains dehydrocostuslactone 0.02mg, promptly.
The preparation of need testing solution: get embodiment 1 and adopt 90% ethanol refluxing process to prepare the Radix Vladimiriae extract 20mg of gained, the accurate methanol 50ml that adds claims to decide weight, reflux, extract, 45 minutes, put coldly, claim to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, get in the measuring bottle that subsequent filtrate 1ml is diluted to 10ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
The chromatogram of dehydrocostuslactone reference substance is seen Fig. 1, and the chromatogram of Radix Vladimiriae extract is seen Fig. 2.
Assay method reproducible test results RSD is 1.19%; Assay method Precision test result RSD is 0.91%; Assay method average recovery rate result of the test is 97.51%, RSD=2.38%; Above result of the test shows that methodology is good, sees Fig. 1 and Fig. 2.
3, the thin layer discrimination method of dehydrocostuslactone and costunolide in the Radix Vladimiriae extract of the present invention
Get embodiment 1 and adopt 90% ethanol refluxing process to prepare the Radix Vladimiriae extract 10mg of gained, add methanol 5ml and make dissolving as need testing solution.Get costunolide respectively, the dehydrocostuslactone reference substance is an amount of, add methanol respectively and make solution that every 1ml contains 0.5mg product solution in contrast.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 B) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose that contains 2.5% silver nitrate, with cyclohexane extraction-ethyl acetate-acetone (8: 1: 1) is developing solvent, launch, take out, dry, spray 5% vanillin sulfuric acid solution, it is clear to be heated to speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.Silver nitrate modification thin layer shows that need testing solution and dehydrocostuslactone, costunolide the same color speckle occurs in same position.Adopt thin-layer developing condition under Radix Vladimiriae item of Chinese Pharmacopoeia version in 2005 simultaneously, the thin layer of pharmacopeia condition shows that need testing solution and dehydrocostuslactone, costunolide the same color speckle occurs in same position.The result shows: can confirm to contain dehydrocostuslactone and costunolide in the Radix Vladimiriae extract of the present invention by the thin layer discrimination method.
Below beneficial effect by pharmacodynamics test proof Radix Vladimiriae extract of the present invention.
The screening test of test example 1 Radix Vladimiriae extract promoting the circulation of QI to relieve pain effective site of the present invention
Radix Vladimiriae is multi-national intersection medication in the area, Sichuan.The tradition medication experience shows that Radix Vladimiriae has the drug effect of promoting the circulation of QI to relieve pain, warming middle-JIAO for easing the stomach, and main application form is single usefulness, takes after mixing it with water and the compatibility other drug is made ball and loose or medicated wine with wine.Pharmacological experimental methods such as this The thesis intestinal advances, stripped myenteron, hot plate method, writhing method promote enterokinesia, gastrointestinal antispasmodic, analgesia drug effect position to screen to Radix Vladimiriae.
1. experiment material
1.1 laboratory animal
Mice: Kunming kind, Chengdu University of Traditional Chinese Medicine's Experimental Animal Center, the laboratory animal quality certification number: the real moving pipe o.11 in river.
Rabbit: the big ear of Japan is white, is provided the laboratory animal quality certification number by Sichuan Academy of Medical Sciences's Experimental Animal Center: SCXK (river 2004-15).
1.2 the preparation of medicinal liquid
Get Radix Vladimiriae medical material Vladimiria souliei (Franch.) Ling var.cinerea Ling, (purchase in Aba state, Sichuan Province Chinese crude drug company limited, the place of production: Xiaojin County, Aba state, Sichuan Province, in October, 2004) make extractum with 75% ethanol percolation, extract with multiple solvent, obtain petroleum ether part, ether position, ethyl acetate extract, water position.The Radix Vladimiriae leaf gets 75% ethanol extract of extractum with 75% ethanol percolation.
1.3 experimental drug
Motilium (batch number: C50126127, the accurate font size of traditional Chinese medicines: H10910003, manufacturer: Xian-Janssen Pharmaceutical Ltd.); Dolantin (5%, product batch number 690101); (lot number is acecoline: 0000420); Atropine sulfate injection (Xuzhou Lai En pharmaceutcal corporation, Ltd, lot number: 011020); Tyrode's solution; Sodium chloride (chemical chemical reagent work of burnt alkali group, lot number: 20000706); Potassium chloride (Chengdu chemical reagent factory, lot number: 990928); Magnesium sulfate (MgSO
47H
2O, 20040902), sodium dihydrogen phosphate (Kingsoft, Chengdu chemical reagent factory, lot number: 20001011), sodium bicarbonate (goes up marine rainbow photoinitiator chemical factory, lot number: 010421) Long Huagongshijichang of Chengdu section, lot number:; Calcium chloride (chemical plant among Deng of Tianjin City Tanggu, lot number: 20000106); Glucose (Sichuan Fei Deli pharmaceutical Co. Ltd, lot number: 040901); Aspirin effervescent tablets (product batch number: 0407002); Glacial acetic acid (Chengdu letter He Huagongshijichang, lot number: 030405).
1.4 experimental apparatus
Magnus' bath (50ml type); BL-410 microcomputer type bio signal is gathered and processing system (Chengdu TME Technology Co., Ltd.); Physiological pressure transducer (YH-4 type, Beijing Astronautics Medical Engineering Inst.); YLS-6A intelligence hot-plate instrument (Shandong Academy of Medical Sciences equipment station).
2. experimental technique and result
2.1 each extract of Radix Vladimiriae is to the influence of mice enterokinesia
Get 130 of white mice, male and female half and half, laboratory rearing 3 days, be divided into 13 groups at random, every group 10, water 24h is can't help in fasting, and the continuous irrigation stomach gives respective concentration medicine group 3 days, behind the last administration 30min, gavage carbon powder suspension (10% active carbon, 10% arabic gum) 0.2ml/ only puts to death behind the 20min, opens the abdominal cavity, separate mesentery, the clip upper end is to pylorus, and the lower end places on glass to the intestinal tube of ileocecum portion, gently that small intestinal is stretching, measuring intestinal tube length with ruler is " small intestinal total length ", and the distance from pylorus to the carbon powder forward position is " carbon powder is at the advance distance of enteral ", calculates carbon powder and advances percentage rate (carbon powder propelling percentage rate (%)=carbon powder advance distance (cm)/small intestinal total length (cm) * 100%.The results are shown in Table 4.
Each extract of table 4 Radix Vladimiriae is to the influence of mice enterokinesia (x ± s)
| Group | Number of animals (only) | Dosage (g (extractum)/Kgd) | Charcoal end propelling degree (%) |
| The blank group | 10 | / | 74.71±17.53 |
| The motilium group | 10 | 5×10 -3 | 99.29±1.83** |
| Leaf extractum group | 10 | 1.05 | 72.15±15.13 |
| The extractum high dose group | 10 | 1.05 | 93.91±8.23** |
| The extractum low dose group | 10 | 0.525 | 83.76±10.38 |
| The petroleum ether high dose group | 10 | 0.24 | 68.02±15.64 |
| The petroleum ether low dose group | 10 | 0.12 | 89.08±10.39* |
Continuous table 4
| The ether high dose group | 9 | 0.399 | 69.59±15.74 |
| The ether low dose group | 9 | 0.1995 | 89.42±9.75* |
| The ethyl acetate high dose group | 10 | 0.24 | 88.63±13.76* |
| The ethyl acetate low dose group | 10 | 0.12 | 77.37±17.12 |
| The water layer high dose group | 10 | 1.05 | 90.34±10.93* |
| The water layer low dose group | 10 | 0.525 | 74.58±29.82 |
Annotate: * and blank group compare, P＜0.05; * and blank group compare, P＜0.01
The result shows: motilium group, extractum high dose group can promote enterokinesia (P＜0.01) fairly obviously, and petroleum ether low dose group, ether low dose group, ethyl acetate high dose group, water layer high dose group can obviously promote enterokinesia (P＜0.05), and other groups are to promoting the enterokinesia effect not obvious.
2.2 each extract of Radix Vladimiriae is to the enhanced influence of rabbit duodenum smooth muscle contraction ability due to the acetylcholine
Adjust Magnus' bath, oxygen generator, pressure transducer and BL-410 microcomputer type bio signal and gather and processing system, make operate as normal, zeroing and calibration.Get rabbit (fasting 12h before the experiment), hitting occipital bone with wooden mallet puts to death, cut the abdominal cavity immediately open, take out one section of duodenum, place the beaker that fills tyrode's solution, fall mesentery along the intestinal wall simple separation, and intestinal contents is rinsed well with tyrode's solution, intestinal tube is cut into the intestinal segment of 1.5～2.5cm, observes the motion of normal myenteron earlier; Adjust myenteron tension force in the Magnus' bath, making relaxes contract stablize 10～30min after, the curve of tracing one section myenteron proper motion then, in Magnus' bath, drip various medicinal liquids (seeing below) again, the easypro curve that contracts of immediate record (is all used tyrode's solution flushing myenteron 2～3 times before dripping medicinal liquid at every turn, fresh body lotion can stop 1～2min and bleed off then, waits to relax to contract to return to before the medication to add group medicinal liquid down again after the level).
The order of administration of acetylcholine modeling: can on a myenteron, finish following experiment.Observe the effect of Radix Vladimiriae extract solution: 1. write down normal value; 2. add acetylcholine solution 25 μ l behind the 60s; 3. add Radix Vladimiriae behind the 30s and extract solution 100 μ l; 4. add Radix Vladimiriae behind the 30s again and extract solution 100 μ l; 5. add Radix Vladimiriae behind the 30s again and extract solution 100 μ l; With tyrode's solution flushing myenteron 2～3 times, fresh body lotion can stop 1～2min and bleed off then, waits that relaxing contracts and return to add group medicinal liquid down again after the level before the medication.Observe atropinic effect (because atropinic inhibitory action is too strong, so the experiment of putting behind): 9. write down normal value; 10. add acetylcholine solution 25 μ l behind the 60s; (11) add atropine 25 μ l behind the 30s; With tyrode's solution flushing myenteron 2～3 times, fresh body lotion can stop 1～2min and bleed off then, waits that relaxing contracts and return to add group medicinal liquid down again after the level before the medication.Experimental result sees Table 5.
Each extract of table 5 Radix Vladimiriae is to (the x ± s) of the enhanced influence of rabbit duodenum smooth muscle contraction ability due to the acetylcholine
| Group | Sample | Drug dose (g/ml) | Mean tension |
| Acetylcholine-normal | 10 | / | 3.63±2.31 |
| Atropine-normal | 10 | 0.00025 | -0.94±0.74** |
| Extractum high dose group-normal | 10 | 1.05 | 1.59±1.4** |
| Extractum low dose group-normal | 10 | 0.35 | 3.81±2.43 |
| Petroleum ether high dose group-normal | 9 | 0.2415 | 1.35±1.84** |
| Petroleum ether low dose group-normal | 9 | 0.0805 | 3.87±1.48 |
| Ether high dose group-normal | 10 | 0.399 | 1.62±1.89** |
| Ether low dose group-normal | 10 | 0.133 | 3.9±2.87 |
| Ethyl acetate high dose group-normal | 10 | 0.2415 | 1.73±1.23* |
| Ethyl acetate low dose group-normal | 10 | 0.0805 | 2.46±1.38 |
| Water layer high dose group-normal | 9 | 1.05 | 2.62±1.87 |
| Water layer low dose group-normal | 10 | 0.35 | 2.79±1.16 |
Annotate: * and blank group compare, P＜0.05; * and blank group compare, P＜0.01.
The result shows, atropine, the petroleum ether high dose group, the extractum high dose group, the fairly obvious inhibition acetylcholine of ether high dose group energy is to the contraction (P＜0.01) of smooth muscle, the ethyl acetate high dose group can obviously suppress the contraction (P＜0.05) of acetylcholine to smooth muscle, and it is not obvious to the contraction of smooth muscle that other groups suppress acetylcholine.
2.3. each extract of Radix Vladimiriae causes the influence that mice licks the metapedes reaction to hot plate
The thermoregulation of hot-plate instrument is caused 55 ± 0.5 ℃, preheating 10min.The record mice from drop into hot plate to the time that metapedes occurs licking (s) as the normal pain threshold of this Mus (survey twice altogether, with meansigma methods greater than 5s and be no more than 30s person for qualified).Get complete female mice, 130, be divided into 13 groups at random, 10 every group; The 1st group is the blank group, gives the equal-volume distilled water at every turn; The 2nd group is the dolantin group, only last lumbar injection 0.2% dolantin 0.2ml/10g; Other each group is respectively 11 extracts of Radix Vladimiriae, measure normal pain threshold after, be subjected to the reagent thing, after the last administration, measure a pain threshold at 60min, surpass 60s as it, promptly stop test and in 60s.Experiment finishes, and presses the threshold of pain raising percentage rate that the pain threshold of being surveyed calculates each time point, and formula is the average pain threshold before threshold of pain improvement value=(the average pain threshold before the pain threshold medication after the medication)/medication.Experimental result sees Table 6.
Table 6 Radix Vladimiriae extract causes the influence that mice licks metapedes reaction (x ± s) to hot plate
Annotate: * and blank group compare, P＜0.05; * and blank group compare, P＜0.01.
The result shows: dolantin dosage group, petroleum ether high dose group, water layer high dose group cause mice to hot plate and lick the metapedes reaction fairly obvious inhibitory action (P＜0.01) is arranged, can obviously prolong the time of occurrence of licking metapedes, the water layer low dose group causes mice to hot plate and licks metapedes reaction obvious suppression effect (P＜0.05) is arranged, can obviously prolong the time of occurrence of licking metapedes, other dosage groups are not obvious to the inhibitory action that mice licks the metapedes reaction.
2.4. each extract Dichlorodiphenyl Acetate of Radix Vladimiriae causes the influence of mouse writhing reaction
Get 130 of mices, be divided into 13 groups at random, 10 every group by body weight; The 1st group is the blank group, gives the equal-volume distilled water every day; The 2nd group is the aspirin group, every day ig aspirin 0.03g/kg; Other groups are respectively 11 extracting solution of Radix Vladimiriae; More than the equal successive administration of each administration group 5 days.Behind the last administration next day 30min, difference lumbar injection 0.6% glacial acetic acid (0.2ml/ is only); Observe and to occur turning round the number of times that occurs writhing response in time (turning round the body latent time) of embodying elephant and the 20min first, the calculating medicine is to the suppression ratio (suppression ratio=(blank group writhing response mean administration group writhing response mean)/blank group writhing response mean) of writhing response.Experimental result sees Table 7.
Each extract Dichlorodiphenyl Acetate of table 7 Radix Vladimiriae causes the influence (x ± s) of mouse writhing reaction
| Group | Number of animals (only) | Drug dose (g/kg) | Turn round body latent time (min) | Writhing response number (inferior) | Suppression ratio (%) |
| The blank group | 10 | / | 4.5±3.03 | 15.6±12.23 | 0 |
| The aspirin group | 10 | 0.03 | 18.3±5.38** | 0.8±2.53** | 95 |
| Leaf extractum group | 10 | 1.5 | 4.4±0.70 | 17.60±11.26 | 0 |
| The extractum high dose group | 10 | 1.05 | 5.5±1.58 | 16.10±10.05 | 0 |
| The extractum low dose group | 10 | 0.525 | 7±4.78 | 10.80±10.91 | 31 |
| The petroleum ether high dose group | 10 | 0.24 | 5.6±5.15 | 10±7.9 | 36 |
| The petroleum ether low dose group | 10 | 0.12 | 4.8±1.03 | 9.2±6.46 | 41 |
| The ether high dose group | 9 | 0.399 | 5±1.83 | 8.75±6.8 | 44 |
| The ether low dose group | 9 | 0.1995 | 5±3.37 | 16.25±8.99 | 0 |
| The ethyl acetate high dose group | 10 | 0.24 | 7.7±6.58 | 7.6±5.3* | 51 |
| The ethyl acetate low dose group | 10 | 0.12 | 6.4±4.9 | 5.9±4.43* | 62 |
| The water layer high dose group | 10 | 1.05 | 6.5±4.91 | 12.30±8.63 | 21 |
| The water layer low dose group | 9 | 0.525 | 4±1.32 | 14.11±7.08 | 10 |
Annotate: * and blank group compare, P＜0.05; * and blank group compare, P＜0.01.
The result shows: the aspirin Dichlorodiphenyl Acetate causes the mouse writhing reaction fairly obvious inhibitory action (P＜0.01), also can prolong fairly obviously and turn round body latent time (P＜0.01); Ethyl acetate high dose group, ethyl acetate low dose group Dichlorodiphenyl Acetate cause the mouse writhing reaction obvious suppression effect (P＜0.05).
3. pharmacological testing The selection result
The medicine efficacy screening result of the test shows that Radix Vladimiriae medical material (root) extracts extractum and each chemical position effect is obvious, and the effect of dis-medicinal part (leaf) extraction extractum is not obvious.
By pharmacodynamics evaluation study to chemical position, petroleum ether, the effect of ether position are obviously, have the contraction of very remarkable inhibition smooth muscle and very significant central analgesia effect, has tangible promotion enterokinesia effect, close with the effect of extraction extractum, identical with the effect of Radix Vladimiriae promoting the circulation of QI to relieve pain.
Each extract promoting the circulation of QI to relieve pain pharmacodynamic analysis (estimating) of table 8 Radix Vladimiriae with significant level
| The position | Test (1) | Test (2) | Test (3) | Test (4) |
| Root extractum position | ** | ** | ||
| Petroleum ether part | * | ** | ** | |
| The ether position | * | ** | ||
| Ethyl acetate extract | * | * | * | |
| The water position | * | ** | ||
| Leaf extractum position |
*: represent utmost point significant level, i.e. P＜0.01; *: represent significant level, i.e. P＜0.05.
4. conclusion
Petroleum ether, ether position are the main effective site that has the effect of promoting the circulation of QI to relieve pain in the Radix Vladimiriae, and carry out chemical constituent at effective site and separate, and obtain the index composition.Above-mentioned test explanation, the ethyl acetate extract effect balance of medicine Radix Vladimiriae of the present invention is weaker than petroleum ether, ether position.Adopt ethyl acetate as refining solvent, can contain petroleum ether, ether position in extraction, safety is higher, has feasibility.
The 2 extract antiulcer tests of the present invention of test example
1. be subjected to the preparation method of reagent thing preparation method and pharmacological testing screening sample:
The Radix Vladimiriae extract: get Radix Vladimiriae and be ground into coarse powder, adding concentration is 85% ethanol, after the infiltration, and reflux, extract,, decompression recycling ethanol adds the water suspendible, ethyl acetate extraction, the reclaim under reduced pressure ethyl acetate adds Polyethylene Glycol-400 and makes the solution that every 1ml contains the 0.5g primary crude drug, and is standby.
2. experiment equipment: slide gauge, magnifier etc.
Experimental drug: Radix Vladimiriae extract, ranitidine hydrochloride capsules (SouthWest Synthetic Pharmaceutical Corp. Ltd, product batch number: 050604), reserpine (Guangdong Bangmin Pharmaceutical Co., Ltd., product batch number: 041027), formaldehyde (Long Huagongshijichang of Chengdu section, product batch number: 20050702)
Laboratory animal: KM mice, male and female half and half, body weight 20g, Chengdu University of Traditional Chinese Medicine's Experimental Animal Center, the laboratory animal quality certification number: the real moving pipe o.11 in river.
3. experimental technique: random packet, every group 10, each organizes the medicine of capacity such as gavaging, continuous 7 days, 1h after the last administration, subcutaneous injection of reserpine 10mg/kg behind the water 24h is can't help in fasting, and animal is put to death in the cervical vertebra dislocation behind the 6h, stomach is got in dissection, the pars cardiaca of ligation stomach and pyloric part also inject 1% formaldehyde 2ml through coat of the stomach in gastral cavity, stomach is taken out immerse in 1% formalin, and 30min tailing edge greater gastric curvature is cut off, the counting ulcer index, the blood stasis point is 1 minute, and wire blood stasis length＜1mm is 2 minutes, and 1-2mm is 3 minutes, 34mm is 4 minutes, is 5 minutes greater than 5mm.
Table 9 screening test table and table as a result
The Radix Vladimiriae extract has tangible ulcer protective effect.
The antiulcer test (comparison being arranged) of test example 3 extracts of the present invention with dehydrocostuslactone
1, experiment material
1.1 laboratory animal
The KM mice: regular grade, male and female half and half, body weight 18～22g is provided by Sichuan Academy of Medical Sciences institute of lab animals, credit number SCXK (river) 2004-15.
The SD rat: the SPF level, male and female half and half, body weight 150～180g is provided by Sichuan Academy of Medical Sciences institute of lab animals, credit number SCXK (river) 2004-15.
Raising condition: herbal pharmacology laboratory animal observation ward of pharmaceutical college of Chengdu University of Traditional Chinese Medicine (credit number: 015).After animal enters observation ward, female, male sub-cage rearing, standby.
1.2 experimental drug
Radix Vladimiriae: (Abazangzuqiangzu Autonomous Prefecture Xiaojin County, Sichuan Province) fixed point purchase in the Radix Vladimiriae main producing region, identify through Chengdu University of Traditional Chinese Medicine's national medicine institute, meet the pertinent regulations under a Radix Vladimiriae Radix of Chinese Pharmacopoeia version in 2005 the vladimiriae item.The ranitidine capsule (SouthWest Synthetic Pharmaceutical Corp. Ltd, lot number: 061003), reserpine injection (Beijing Double-Crane Pharmaceutical Co., Ltd company limited, lot number: 051027).
1.3 experiment reagent
Glacial acetic acid (Long Huagongshijichang of Chengdu section), 40% formalin (Long Huagongshijichang of Chengdu section), PEG400 (Long Huagongshijichang of Chengdu section), above reagent is analytical pure.
1.4 experiment equipment
Tweezers, operating scissors, surgical thread, operation needle, microsyringe, syringe needle, cotton swab, electronic balance, filling stomach syringe needle, syringe, mosquito forceps, slide gauge, magnifier.
1.5 test sample preparation and dosage setting
Preparation of model control group test sample and dosage setting: replace medicine with moisture Polyethylene Glycol-400, equal capacity gastric infusion.
Preparation of ranitidine test sample and dosage setting: get the ranitidine capsule 's content, add moisture Polyethylene Glycol-400 and make the solution (dosage is 50mg/kg) that every 1ml contains 5mg, standby.
Preparation of dehydrocostuslactone test sample and dosage setting: get Radix Vladimiriae, be ground into coarse powder, adding concentration is 90% ethanol, after the infiltration, reflux, extract,, decompression recycling ethanol adds the water suspendible, defat with petroleum ether, extracted with diethyl ether reclaims solvent, adds silica gel and mixes thoroughly, silica gel column chromatography, cyclohexane extraction-ethyl acetate gradient elution, collecting ring hexane-ethyl acetate (95: 5) eluent, decompression and solvent recovery, crystallization, cyclohexane extraction-ether recrystallization is collected colourless column crystallization, gets colourless column crystallization, add moisture Polyethylene Glycol-400 and make high dose group test sample (dosage is 90mg/kg), middle dosage group test sample (dosage is 45mg/kg), low dose group test sample (dosage is 15mg/kg), standby.
Preparation of ethyl acetate extract test sample and dosage setting: get Radix Vladimiriae, be ground into coarse powder, adding concentration is 90% ethanol, reflux, extract,, decompression recycling ethanol adds the water suspendible, ethyl acetate extraction, the reclaim under reduced pressure ethyl acetate adds moisture Polyethylene Glycol-400 and makes high dose group test sample (dosage is 9g/kg), low dose group test sample (dosage is 4.5g/kg), and is standby.
Preparation of ethanol extraction test sample and dosage setting: get Radix Vladimiriae, be ground into coarse powder, adding concentration is 90% ethanol, reflux, extract,, and decompression recycling ethanol adds moisture Polyethylene Glycol-400 and makes high dose group test sample (dosage is 9g/kg), and is standby.
2, experimental technique
2.1 Radix Vladimiriae causes the influence of the experimental gastric ulcer of chmice acute to reserpine
Get 80 of KM mices, divide 8 groups of (model control group at random, the ranitidine matched group, the dehydrocostuslactone height, in, low dose group, the ethyl acetate extract height, middle dosage group, the ethanol extraction high dose group), every group 10, each organizes gastric infusion 0.2ml/20g, model control group gives the moisture Polyethylene Glycol of equivalent, continuous 5 days, water 24h is can't help in fasting before the experiment, each organizes administration again 1 time, subcutaneous injection of reserpine 6mg/kg behind the 1h, animal is put to death in the cervical vertebra dislocation behind the 6h, stomach is got in dissection, the pars cardiaca of ligation stomach and pyloric part also inject 1% formaldehyde 2ml through coat of the stomach in gastral cavity, stomach is taken out in immersion 1% formalin, 30min tailing edge greater gastric curvature is cut off, clean gastric content, gastric mucosa is launched, and the strip that gastric mucosa injury shows as glandular stomach district mucosa is hemorrhage, measures the degree of gastric mucosa injury with slide gauge, (point system: not damaged is 0 minute, the blood stasis point is 1 minute, and wire blood stasis length＜1mm is 2 minutes, and 1～2mm is 3 minutes, 2～3mm is 4 minutes, greater than 3mm is 5 minutes), calculate ulcer index, and calculate ulcer inhibition rate.
2.2 the Radix Vladimiriae Dichlorodiphenyl Acetate causes the influence of the experimental gastric ulcer of rat chronic
Get 40 of SD rats, behind the fasting 12h, the stomach wall hair is cut in lumbar injection 5% chloral hydrate 2ml anesthesia, with iodine tincture and alcohol disinfecting.2～2.5cm place cuts off skin and stomach wall in xiphoid-process hypogastric region center, with curved hemostat below the liver of right side with duodenum gently hook go out, pull out glandular stomach portion outside the abdomen gently then, facies ventralis at stomach, microsyringe is put down 0.4～0.5mm place under the serous coat that thrusts body of stomach and pyloric antrum intersection, annotate people 40% acetic acid 0.05ml, sew up the incision, the postoperative routine feeding, divide 4 groups (model control group, ranitidine matched group, ethyl acetate extract height, middle dosage groups) next day at random, every group 10, every day gastric infusion 2ml/200g, successive administration 14 days.Put to death animal on the 15th day, take out behind the stomach with the fixing 10min of 1% formalin, cut off coat of the stomach along greater gastric curvature, stomach is turned up, clean gastric content, get the glandular stomach district and launch, blot moisture in the ulcer with paper, measure the degree of gastric mucosa injury with slide gauge, with the equal value representation ulcer index of the major diameter of ulcer and minor axis, and the calculating ulcer inhibition rate.
3, date processing and result judge
Experimental data is with mean+SD (x ± s) expression, the one factor analysis of variance method variance analysis (Homogeneity of variance/ANOVA/Test of Homogeneity of Variances) of adopting SPSS software to provide, program by variance analysis is carried out homogeneity test of variance earlier: homogeneity of variance, add up with the comparative approach in twos (Multiple Comparisons/LSD method) of mean between a plurality of experimental grouies and matched group; Heterogeneity of variance carries out the conversion of suitable variable to data, wait to satisfy normal state or variance are neat require after, add up with the data after changing; If do not reach homogeneity of variance yet after the variable conversion, use rank test instead and add up.
3.1 Radix Vladimiriae causes the inhibitory action of the experimental gastric ulcer of chmice acute to reserpine
Data statistic analysis is the result show: compare with model control group, Radix Vladimiriae extract and effective ingredient thereof have highly significant inhibition reserpine and cause the experimental gastric ulcer formation of chmice acute effect (P＜0.01) (table 10, Fig. 3).
Data statistic analysis is the result also show: Radix Vladimiriae extract and effective ingredient thereof, and suppress reserpine and cause the experimental gastric ulcer formation of chmice acute effect, close with the ranitidine matched group, do not have significant differences (P〉0.05); Ethyl acetate extract high dose group wherein, the ulcer inhibition rate significance is higher than ranitidine matched group (P＜0.05); Each dosage group of ethyl acetate extract wherein, each dosage group of ulcer inhibition rate and dehydrocostuslactone relatively do not have significance meaning (P〉0.05).
Each test dose group and model control group compare; ulcer inhibition rate is higher than model control group; and statistical procedures difference has the significance meaning, can judge that Radix Vladimiriae extract and effective ingredient thereof cause the experimental gastric ulcer of chmice acute to reserpine defencive function is arranged.
Wherein the effect of ethyl acetate extract high dose group is obvious, and ulcer inhibition rate is higher than each dosage group of dehydrocostuslactone, is higher than ethanol position dosage group simultaneously, also is higher than the ranitidine matched group simultaneously.
Table 10 Radix Vladimiriae causes the inhibitory action result of the test of the experimental gastric ulcer of chmice acute to reserpine
| Group | Number of animals only) | Drug dose | Ulcer index (x ± s) | Ulcer inhibition rate (%) |
| 1. model control group | 10 | 0 | 31.57±9.69 | 0 |
| 2. ranitidine matched group | 10 | 50mg/kg | 19.78±7.76** | 37.36 |
| 3. dehydrocostuslactone high dose group | 10 | 90mg/kg | 16.22±7.76** | 48.62 |
| 4. dosage group in the dehydrocostuslactone | 10 | 45mg/kg | 13.56±8.05** | 57.06 |
| 5. dehydrocostuslactone low dose group | 10 | 15mg/kg | 14.89±7.54** | 52.84 |
| 6. ethyl acetate extract high dose group 1 | 10 | 9g/kg | 11.33±10.81**△ | 64.10 |
| 7. dosage group in the ethyl acetate extract 1 | 10 | 4.5g/kg | 18.22±9.44** | 42.28 |
| 8. ethanol position high dose group 1 | 10 | 9g/kg | 12.36±8.26** | 60.84 |
Compare with model control group: * P＜0.05; * P＜0.01; Compare with ranitidine contrast dosage group: Δ P＜0.05
1: ethyl acetate extract, ethanol position, the pharmaceutical dosage unit of its each dosage group is (a g primary crude drug/kg mice body weight), can extract extract 60～80g of the present invention approximately according to calculating 1kg Radix Vladimiriae medical material, 1kg Radix Vladimiriae medical material can extract dehydrocostuslactone monomer 10～30g approximately, dehydrocostuslactone content is suitable in extract high dose group of the present invention as can be known and the dehydrocostuslactone high dose group of converting, in the extract of the present invention in dosage group and the dehydrocostuslactone in the dosage group dehydrocostuslactone content suitable.
In the actual production, the extract position that 1g Radix Vladimiriae medical material can prepare: ethanol position (the present invention only adopts alcohols solvent to extract dry solvent and partly gets extract) 240mg (yield about 24%); Ethyl acetate extract (being extract of the present invention) 60mg (yield 6%); Dehydrocostuslactone 10mg (yield 1%)
3.2 the Radix Vladimiriae Dichlorodiphenyl Acetate causes the inhibitory action of the experimental gastric ulcer of rat chronic
Experimental result shows: ethyl acetate extract (high dose group), and suppress acetic acid and cause the experimental gastric ulcer formation of rat chronic effect, have highly significant meaning (comparing P＜0.01) with model control group; Ethyl acetate extract (middle dosage group), inhibition acetic acid cause the experimental gastric ulcer formation of rat chronic effect and have significance meaning (comparing P＜0.05 with model control group); Ranitidine contrast dosage group, inhibition acetic acid cause the experimental gastric ulcer formation of rat chronic effect and have highly significant meaning (comparing P＜0.01 with model control group) (table 11, Fig. 4).
Data statistic analysis is the result also show: Radix Vladimiriae extract and effective ingredient thereof, and suppress reserpine and cause the experimental gastric ulcer formation of chmice acute effect, compare with the ranitidine matched group, have significant differences (P＜0.01).
Each test dose group and model control group compare; ulcer inhibition rate is higher than model control group; and statistical procedures difference has the significance meaning, and Radix Vladimiriae extract Dichlorodiphenyl Acetate causes the experimental gastric ulcer of rat chronic defencive function, and wherein the effect of ethyl acetate extract high dose group is obvious.
Table 11 Radix Vladimiriae Dichlorodiphenyl Acetate causes the inhibitory action result of the test of the experimental gastric ulcer of rat chronic
| The group preface | Sample number (only) | Drug dose | Ulcer index (x ± s) | Ulcer inhibition rate (%) |
| 1. model control group | 10 | 0 | 4.69±1.28△△ | 0 |
| 2. ranitidine contrasts the dosage group | 10 | 50mg/kg | 1.29±1.10** | 72.43 |
| 3. ethyl acetate extract high dose group 1 | 10 | 9g/kg | 3.17±0.70**△△ | 32.42 |
| 4. dosage group in the ethyl acetate extract 1 | 10 | 4.5g/kg | 3.64±0.93*△△ | 22.28 |
Compare with model control group: * P＜0.05; * P＜0.01
Compare with ranitidine contrast dosage group: Δ P＜0.05; Δ Δ P＜0.01
1: ethyl acetate extract, the pharmaceutical dosage unit of its each dosage group are (g primary crude drug/kg mice body weight)
4, discuss
4.1 ranitidine is as typical H2 acceptor inhibitor, in the treatment of Peptic Ulcers, good curative effect is arranged, mainly by gastric acid inhibitory secretion, reduce acidity and suppress ulcer, from this research the data of ulcer inhibition rate as can be seen the effect of Dichlorodiphenyl Acetate type gastric ulcer be better than the reserpine type.
According to traditional medication custom and early-stage Study; the antiulcer action that compares ranitidine; the Radix Vladimiriae ethyl acetate extract is better than acetic acid type to the effect of reserpine type gastric ulcer; may be to the protective effect of reserpine type ulcer with to improve mechanism of action such as gastrointestinal motility and stomachial secretion relevant; being suitable for the abdominal distention gastric abscess is the Peptic Ulcers of cardinal symptom, reaches the antiulcer effect with promoting the circulation of QI to relieve pain (abdominal distention → gastric abscess → gastric ulcer).
From the clinical practice performance analysis, adopt Radix Vladimiriae and preparation determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs on the basis of traditional medication experience thereof, in case of necessity as the adjuvant drug of chemotherapy, avoid long-term but sour treatment back drug withdrawal to cause the rising of gastric acid secretion bounce-back property, reduce the recurrence of ulcer disease rate, reduce untoward reaction, increase the effectiveness of clinical practice.
4.2 from the experimental data statistic analysis result as can be seen: to reserpine type gastric ulcer, ethyl acetate extract and other ulcer index of dehydrocostuslactone respective sets do not have significant difference; Dichlorodiphenyl Acetate type gastric ulcer compares with model control group, and the ulcer inhibition rate of Radix Vladimiriae ethyl acetate extract high dose group is higher, has the statistical significance of highly significant.
Radix Vladimiriae ethyl acetate extract preparation method is simple, preparation cost is far below unification compound (dehydrocostuslactone), under equal dosage, the cost effect is with the obvious advantage, the value of furtheing investigate and applying is arranged as the Radix Vladimiriae effective extract, this paper result of study will be better, cheap for seeking curative effect, and the Mucosta thing of few side effects provides the experimental basis of Application and Development.
In sum, the Radix Vladimiriae extract has clear and definite antiulcer action and promoting the circulation of QI to relieve pain effect, by illustrating the traditional medication experience and the mechanism of action thereof of Radix Vladimiriae, in conjunction with the domestic and foreign literature research data, will find to provide the research thinking for the new drug of Peptic Ulcers.
Adopt extract of the present invention can be prepared as the preparation of following form:
1, the preparation of medicinal dropping ball of the present invention: get Radix Vladimiriae extract 200g, add in the 800g poloxamer Polyethylene Glycol-6000 (1:3) of heating and melting, stir, be transferred in the drop pill machine under 60-70 ℃ of heat-retaining condition, splash in 10 ℃ the dimethicone, collect the drop pill of molding, removing oil, make the 1000g drop pill, promptly.
Oral, three times on the one, each consumption 0.25g, every 0.25g contains Radix Aucklandiae extract 50mg.
2, the preparation of medicine capsule of the present invention: get Radix Vladimiriae extract 50g, beta cyclodextrin 250g, adopt polishing to carry out enclose, inclusion complex is loaded into 1000 capsules, promptly.
Oral, three times on the one, 1 of each consumption, per 1 contains Radix Aucklandiae extract 50mg.
3, the preparation of medicinal soft capsule agent of the present invention: get Radix Vladimiriae extract 50g, add 200g soybean oil, 10g tween 80,40g Cera Flava, mix homogeneously in the homogenizer is pressed into 1000 soft capsules, promptly.
Oral, three times on the one, 1 of each consumption, per 1 contains Radix Vladimiriae extract 50mg.
4, the preparation of medicinal tablet of the present invention: get Radix Vladimiriae extract 50g, beta cyclodextrin 500g, adopt polishing to carry out enclose, inclusion complex adds starch 50g, and mixing is made granule, is pressed into 1000, the bag film-coat, promptly.
Oral, three times on the one, 1 of each consumption, per 1 contains Radix Aucklandiae extract 50mg.
Claims (10)
- [claim 1] Radix Vladimiriae extract is characterized in that, it is to be raw material with the medical material Radix Vladimiriae, and supercritical extraction or add after alcohols solvent extracts adds aqueous suspension, adopts the non-polar organic solvent extract and separate, collects organic facies and partly is drying to obtain.
- [claim 2] Radix Vladimiriae extract according to claim 1 is characterized in that described alcohols solvent is an ethanol, and concentration of alcohol is 30%～95%; Described non-polar organic solvent be in ether, petroleum ether or the ethyl acetate any one.
- [claim 3] Radix Vladimiriae extract according to claim 2 is characterized in that described concentration of alcohol is 70～95%.
- [claim 4] Radix Vladimiriae extract according to claim 1 is characterized in that, it is by following method preparation:A, take by weighing Radix Vladimiriae and pulverize, adopt supercritical extraction method, solvent supersonic extraction method, solvent percolation extraction method or solvent refluxing extraction method to extract, solvent is an alcohols solvent;B, with the solvent extraction liquid of a step preparation, behind the reclaim under reduced pressure alcohols solvent, add aqueous suspension, adopt the non-polar organic solvent extract and separate, behind the reclaim under reduced pressure non-polar organic solvent, drying under reduced pressure organic facies part promptly gets Radix Vladimiriae extract of the present invention again.
- [claim 5] prepares the method for the described Radix Vladimiriae extract of claim 1, it is characterized in that, it is by following method preparation:A, take by weighing Radix Vladimiriae and pulverize, adopt supercritical extraction method, solvent supersonic extraction method, solvent percolation extraction method or solvent refluxing extraction method to extract, solvent is an alcohols solvent;B, with the solvent extraction liquid of a step preparation, behind the reclaim under reduced pressure alcohols solvent, add aqueous suspension, adopt the non-polar organic solvent extract and separate, behind the reclaim under reduced pressure non-polar organic solvent, drying under reduced pressure organic facies part promptly gets Radix Vladimiriae extract of the present invention again.
- The purposes of the described Radix Vladimiriae extract of [claim 6] claim 1 in the medicine of preparation treatment Peptic Ulcers.
- The described Radix Vladimiriae extract of [claim 7] claim 1 is purposes in the medicine of Peptic Ulcers of cardinal symptom with the abdominal distention gastric abscess in preparation treatment.
- [claim 8] a kind of pharmaceutical composition, it is that Radix Vladimiriae extract with each described effective dose of claim 1-4 is an active component, adds the medicament that acceptable accessories or complementary composition are prepared from.
- [claim 9] pharmaceutical composition according to claim 8 is characterized in that: described medicament is capsule, soft capsule, tablet, granule, powder, drop pill, oral liquid or pill.
- The purposes of the described pharmaceutical composition of [claim 10] claim 8 in the medicine of preparation treatment Peptic Ulcers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710202833 CN101450092B (en) | 2007-12-03 | 2007-12-03 | Vladimiria root extract, composition containing the vladimiria root extract and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710202833 CN101450092B (en) | 2007-12-03 | 2007-12-03 | Vladimiria root extract, composition containing the vladimiria root extract and use thereof |
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| Publication Number | Publication Date |
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| CN101450092A true CN101450092A (en) | 2009-06-10 |
| CN101450092B CN101450092B (en) | 2013-01-09 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200710202833 Expired - Fee Related CN101450092B (en) | 2007-12-03 | 2007-12-03 | Vladimiria root extract, composition containing the vladimiria root extract and use thereof |
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| Country | Link |
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| CN (1) | CN101450092B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102133247A (en) * | 2011-01-28 | 2011-07-27 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN102716169A (en) * | 2011-07-01 | 2012-10-10 | 成都中医药大学 | Application of common vladimiria root or extract of common vladimiria root in preparing medicines for treating chronic atrophic gastritis |
| CN103784489A (en) * | 2012-10-30 | 2014-05-14 | 中国医学科学院药用植物研究所 | Banksia rose extract product, preparation method and application thereof |
| CN110353689A (en) * | 2019-07-19 | 2019-10-22 | 淄博职业学院 | A kind of writhing assay monitoring device |
| CN110946889A (en) * | 2019-12-27 | 2020-04-03 | 西南大学 | Application of radix aucklandiae or radix aucklandiae extract in preparation of anti-cholestasis drug |
| CN117045563A (en) * | 2023-09-28 | 2023-11-14 | 云南英格生物技术有限公司 | Radix aucklandiae root extract and its preparation method and application |
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| CN100333772C (en) * | 2005-01-25 | 2007-08-29 | 蒋迎春 | Medicine for treating digestive tract ulcer and stomach cercinoma prophase pathologic change |
| CN101007044B (en) * | 2006-01-23 | 2010-06-09 | 成都中医药大学 | A medicine composition for treating peptic ulcer, preparing method and use thereof |
| CN104848325B (en) * | 2015-04-24 | 2018-09-11 | 珠海格力电器股份有限公司 | Absorption heat pump type heat exchanger unit |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102133247A (en) * | 2011-01-28 | 2011-07-27 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN102133247B (en) * | 2011-01-28 | 2013-03-13 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN102716169A (en) * | 2011-07-01 | 2012-10-10 | 成都中医药大学 | Application of common vladimiria root or extract of common vladimiria root in preparing medicines for treating chronic atrophic gastritis |
| CN102716169B (en) * | 2011-07-01 | 2014-07-30 | 成都中医药大学 | Application of common vladimiria root or extract of common vladimiria root in preparing medicines for treating chronic atrophic gastritis |
| CN103784489A (en) * | 2012-10-30 | 2014-05-14 | 中国医学科学院药用植物研究所 | Banksia rose extract product, preparation method and application thereof |
| CN110353689A (en) * | 2019-07-19 | 2019-10-22 | 淄博职业学院 | A kind of writhing assay monitoring device |
| CN110946889A (en) * | 2019-12-27 | 2020-04-03 | 西南大学 | Application of radix aucklandiae or radix aucklandiae extract in preparation of anti-cholestasis drug |
| CN117045563A (en) * | 2023-09-28 | 2023-11-14 | 云南英格生物技术有限公司 | Radix aucklandiae root extract and its preparation method and application |
| CN117045563B (en) * | 2023-09-28 | 2023-12-29 | 云南英格生物技术有限公司 | Radix aucklandiae root extract and its preparation method and application |
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| CN101450092B (en) | 2013-01-09 |
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