CN1686241A - Tupistra Chinensis Bak extract medicinal composition, and its preparation method and use same - Google Patents
Tupistra Chinensis Bak extract medicinal composition, and its preparation method and use same Download PDFInfo
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- CN1686241A CN1686241A CN 200510020677 CN200510020677A CN1686241A CN 1686241 A CN1686241 A CN 1686241A CN 200510020677 CN200510020677 CN 200510020677 CN 200510020677 A CN200510020677 A CN 200510020677A CN 1686241 A CN1686241 A CN 1686241A
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Abstract
An extract of tupistra Chinensis Bark, which contains its water-soluble and liposoluble components and its saponin, a medicine containing said extract, and its preparing process and application are disclosed.
Description
Technical field
The present invention relates to a kind of Tupistra chinensis Bak. extract, particularly, is the extract with the natural drug feedstock production, the invention still further relates to contain the pharmaceutical composition that this extract is an active component, belongs to drug world.
Background technology
Pharyngolaryngitis is a kind of commonly encountered diseases, frequently-occurring disease clinically, belongs to upper respiratory disease, is the inflammatory lesion of bottleneck throat mucous membrane tissue, is often brought out by suffer from cold, fatigue etc., invades the mucosa of bottleneck throat and causes with antibacterial, virus.Time according to morbidity is different with symptom, can be divided into acute pharyngolaryngitis and chronic pharyngolaryngitis.Acute pharyngolaryngitis is a kind of acute inflammation that is caused by antibacterial or virus, and chronic pharyngolaryngitis is the chronic nonspecific inflammation of bottleneck throat; And acute pharyngolaryngitis shows effect repeatedly and can change chronic pharyngolaryngitis into.Studies show that inflammation generally was divided into for three phases: inflammation is early stage, claim the vascular reaction phase again, show as telangiectasis and permeability increases, ooze out edema; Inflammation mid-term claims the cell effect phase again, shows as leukocyte and gathers to the inflammatory position; Inflammation late period claims tissue reaction's phase again, shows as proliferation of fibrous tissue, and the inflammation barrier forms.This shows that the numerous and pathogenesis of the cause of disease of pharyngolaryngitis relates to a plurality of aspects, a plurality of link, thereby the prompting Drug therapy should play comprehensive treatment by multipath.
Tupistra chinensis Bak. (Tupistra Chinensis Bak.) is a Liliaceae Herba Convallariae family Tupistra chinensis Bak. platymiscium, and its former plant has another name called Rhizoma Belamcandae, Rohdea japonica Roth.The record of Chinese medicine voluminous dictionary, the Tupistra chinensis Bak. sweet and slightly bitter taste, cold in nature, cure mainly diseases such as consumptive fever cough, traumatic injury, rheumatic arthralgia, menoxenia
[1]Shaanxi Chinese herbal medicine record, Tupistra chinensis Bak. removes rheumatism, clearing away heat-fire, pain easing and hemostasis, regulating menstruation and activating blood, YIN nourishing tonify deficiency.Treatment rheumatic arthritis, lumbago and skelalgia, traumatic injury, impairment caused by overstrain, menoxenia, hectic fever due to YIN-deficiency consumptive fever
[1].Chengdu Chinese herbal medicine record, Tupistra chinensis Bak. benefiting QI for activating blood circulation, lung heat clearing, detoxifcation
[1]Hubei Chinese herbal medicine will record, the cold of Tupistra chinensis Bak. nature and flavor sweetness and bitterness, poisonous.Have lung heat clearing, sore-throat relieving, invigorate blood circulation, the analgesic effect, cure mainly diseases such as headache, throat pain, lumbar and back pain, arthralgia, cough, traumatic injury, burn, scald
[2]The sweet peaceful blue or green Chinese herbal medicine choosing record in Shan, the Tupistra chinensis Bak. mildly bitter flavor, cold in nature, slightly poisonous.Have dispel the wind, the effect of pain relieving, dissipating blood stasis.Be used for diseases such as rheumatic arthritis, arthralgia, traumatic injury, hectic fever due to YIN-deficiency consumptive disease heat
[3]Whole nation Chinese herbal medicine compilation record, the Tupistra chinensis Bak. nature and flavor are sweet, little hardship, cold, poisonous.Effect with heat-clearing and toxic substances removing, eliminating stasis to stop pain.Cure mainly diseases such as diphtheria, rheumatic arthritis, lumbago and skelalgia, traumatic injury, lyssodexis
[4]Its collutory treatment pharyngolaryngitis of Tujia and Shennongjia usefulness among the people, tonsillitis, evident in efficacy
[5]The chemical constituent of chloroform extract has had report in the Tupistra chinensis Bak. methanolic extract
[6-8]The structure of having identified is mainly steroid sapogenin, flavone, pterocarpin etc.The chemical constituent of other extract part of Tupistra chinensis Bak. and the compatibility of extract part use and do not appear in the newspapers as yet.
Summary of the invention
Technical scheme of the present invention provides a kind of Tupistra chinensis Bak. extract, and another technical scheme of the present invention is pharmaceutical composition that contains the Tupistra chinensis Bak. extract and its production and use.
The invention provides Tupistra chinensis Bak. Tupistra Chinensis Bak. extract, it contains the component of following weight proportion:
0～10 part of Rhizoma Tupistrae Chinensis aqueous extract, 0～10 part of Rhizoma Tupistrae Chinensis lipoclastic, 1～10 part of Rhizoma Tupistrae Chinensis saponin.
Further, it contains the component of following weight proportion:
10 parts of Rhizoma Tupistrae Chinensis aqueous extracts, 10 parts of Rhizoma Tupistrae Chinensis lipoclastics, 3 parts of Rhizoma Tupistrae Chinensis saponins.
Wherein, described Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, saponin are by following method preparation:
With Liliaceae Herba Convallariae family Tupistra chinensis Bak. platymiscium Tupistra chinensis Bak. Tupistra Chinensis Bak. rhizome is raw material, is solvent with methanol, extracts the dry dark-brown extractum that gets, and is methanolic extract; This extract and distilled water are with 1: 1 mixing of volume ratio, earlier with chloroform extraction, the dry Rhizoma Tupistrae Chinensis lipoclastic that gets; Aqueous solution after the extraction is with water saturated n-butanol extraction, dry Rhizoma Tupistrae Chinensis saponin; The dry Rhizoma Tupistrae Chinensis aqueous extract that gets of remaining aqueous solution.
Wherein, though described Rhizoma Tupistrae Chinensis saponin may be fat-soluble or water solublity, by above-mentioned preparation method as can be known, Rhizoma Tupistrae Chinensis aqueous extract of the present invention, lipoclastic are meant the hydrotrope and the lipoclastic that does not contain saponin.
The present invention also provides a kind of pharmaceutical composition, and it is to be mixed into active component by one of Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, saponin or its, adds acceptable accessories or complementary composition, the preparation that is prepared from.
Wherein, the HPLC finger printing of this pharmaceutical composition is made up of 5 characteristic peaks as shown in Figure 1,
Wherein chromatographic condition is: chromatographic column YMC-Pack ODS-AQ, s-6um, 120,250mm * 4.6mmI.D.; Mobile phase is acetonitrile-K
2HPO
4/ KH
2PO
4, gradient buffer, wherein K
2HPO
4/ KH
2PO
4Concentration is 1mmol/L, and pH value is 5.83, acetonitrile and K
2HPO
4/ KH
2PO
4The volume ratio initial value be 20: 80,10min is 22: 78,15min is 25: 75,20min is 28: 72,34min is 34: 66,44min is 43: 57,48min is 80: 20,52min is 85: 15,60min is 20: 80; Flow velocity is 1.0ml/min; Column temperature is 40 ℃; The detection wavelength is 203nm.
The retention time RT value of described 5 characteristic peaks is respectively: No. 1 peak: 23.2min ± 0.86; No. 2 peak: 24.6min ± 0.90; No. 3 peak: 27.2min ± 0.84; No. 4 peak: 30.4min ± 0.96; No. 5 peak: 33.8min ± 0.98.
The present invention also provides this preparation of drug combination method, comprises the steps:
The preparation of a, Tupistra chinensis Bak. extract: with Liliaceae Herba Convallariae family Tupistra chinensis Bak. platymiscium Tupistra chinensis Bak. TupistraChinensis Bak. rhizome is raw material, is solvent with methanol, extracts the dry dark-brown extractum that gets, and is methanolic extract; This extract with and distilled water with 1: 1 mixing, earlier with chloroform extraction, dry Rhizoma Tupistrae Chinensis lipoclastic; Aqueous solution after the extraction is with water saturated n-butanol extraction, dry Rhizoma Tupistrae Chinensis saponin; The dry Rhizoma Tupistrae Chinensis aqueous extract that gets of remaining aqueous solution;
B, Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, the saponin of getting a step preparation are active component, press column weight amount proportioning and add acceptable accessories or complementary composition, the preparation that is prepared from: 0～10 part of Rhizoma Tupistrae Chinensis aqueous extract, 0～10 part of Rhizoma Tupistrae Chinensis lipoclastic, 1～10 part of Rhizoma Tupistrae Chinensis saponin.
The present invention also provides the purposes of this pharmaceutical composition in the preparation anti-inflammatory drug.Particularly, described medicine is the medicine of treatment pharyngitis.Further, described medicine be suck, oral drugs, injectable drug.
The present invention also provides the purposes of this pharmaceutical composition in the preparation expelling phlegm drugs.
This experiment employing Tupistra chinensis Bak. water and methanolic extract carry out antiinflammatory and test to such an extent that methanolic extract has antiinflammatory curative effect preferably; Because Chinese medicine ingredients complexity, in order further to understand the antiphlogistic active site of Tupistra chinensis Bak., thus the further separation and purification of Tupistra chinensis Bak. methanolic extract is got aqueous solution part (being water soluble ingredient), chloroform part (being liposoluble constituent), n-butyl alcohol part (being the saponin part), confirm that by preliminary experiment the antiphlogistic active site of Tupistra chinensis Bak. is saponin part (the molish reaction is positive); In addition, acute toxicity testing confirms that Rhizoma Tupistrae Chinensis saponin toxicity is minimum, is equivalent to the day for human beings 100 times with dosage, thereby has guaranteed clinical application safety; Adopt pharyngeal spraying ammonia to set up the activity of acute pharyngitis animal model evaluation Tupistra chinensis Bak. treatment acute pharyngitis, the result shows, large ear rabbit pharyngeal pathology and ultrastructure are obviously improved after the Rhizoma Tupistrae Chinensis saponin treatment, and have significance (P＜0.01) with Herba Pileae Scriptae group comparing difference.Bacteriostatic experiment shows that Rhizoma Tupistrae Chinensis saponin has good in-vitro bacteriostasis (MIC:0.063) to bottleneck throat common pathogen such as staphylococcus aureus, staphylococcus epidermidis etc.But in DNCB induced mice DTH,, the inductive mouse DTH of DNCB is not made significant difference (P＞0.05) with continuous 8 days of Rhizoma Tupistrae Chinensis saponin ig administration, the anti-allergic effects that Tupistra chinensis Bak. is described a little less than.Adopting the phenol red excretion method of trachea section to estimate in the expectorant activity of Tupistra chinensis Bak., the phenol red excretion amount of the high, medium and low dosage group of Tupistra chinensis Bak. trachea section is that the OD value is high than the NS group, and significant differences (P＜0.001) is arranged, and NH
4The Cl group more only has significant difference (P＜0.05) than the NS group.Day for human beings clothes dosage is saponin 5～20mg.Each 5～10mg, 2～3 times on the one.Taking dose is little.Rhizoma Tupistrae Chinensis aqueous extract, Rhizoma Tupistrae Chinensis lipoclastic, Rhizoma Tupistrae Chinensis saponin have all been brought into play drug effect, have the effect of Synergistic, and safety, effectively, controllability is strong, stable, provides a kind of new selection for clinical.
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 is 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.
Description of drawings
Five kinds of main saponin constituents and retention time thereof in 70% eluate of Fig. 1 Tupistra chinensis Bak.
The specific embodiment
The preparation of embodiment 1 Tupistra chinensis Bak. extract of the present invention:
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water are fully rotated mixing with volume ratio at 1: 1, dry that brown extractum (fat-soluble) is preserved in refrigerator with chloroform extraction earlier, aqueous solution after the extraction is dry that extractum (saponin) is standby in the freezing preservation of refrigerator with water saturated n-butanol extraction, the dry aqueous solution part that gets of remaining aqueous solution.In the extracting method of three effective kind parts, leaching process, temperature, solvent for use etc. are that very important parameter is as follows:
1, extracts solvent: water, 0-100% methanol aqueous solution, 0-100% ethanol water.
2, extracting mode: except the said extracted method, can use replacements such as carbon dioxide supercritical fluid extraction, solvent extraction or ultrasonic hydrotropy extraction for steroid sapogenin.
Temperature control:
| Extracting mode | Temperature parameter (℃) |
| Water | ????30-100℃ |
| The 0-100% methanol aqueous solution | ????30-70℃ |
| The 0-100% ethanol water | ????30-82℃ |
| Ultrasonic hydrotropy extraction | Room temperature |
| Extracting solution thickening temperature scope | Room temperature-65 ℃ |
| The medicinal material drying temperature range | Room temperature-65 ℃ |
The preparation of embodiment 2 medicine Rhizoma Tupistrae Chinensis saponins of the present invention
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water with 1: 1 mixing of volume ratio, are got brown extractum with chloroform extraction earlier and preserve in refrigerator, and the aqueous solution after the extraction is dry must yellow extractum standby in the freezing preservation of refrigerator with water saturated n-butanol extraction.
The preparation of embodiment 3 medicinal tablets of the present invention
Rhizoma Tupistrae Chinensis aqueous extract 10kg, the Rhizoma Tupistrae Chinensis lipoclastic 10kg, the Rhizoma Tupistrae Chinensis saponin 3kg that get embodiment 1 preparation mix.Get mixture, add starch and 1% magnesium stearate, tabletting makes every to contain saponin 5mg.(used adjuvant can be one or more in tablets such as microcrystalline Cellulose, modified starch, ethyl cellulose, methylcellulose, hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, the sodium carboxymethyl cellulose adjuvant commonly used).
The preparation of embodiment 4 medicine capsules of the present invention
Get Rhizoma Tupistrae Chinensis aqueous extract 8kg, Rhizoma Tupistrae Chinensis lipoclastic 5kg, Rhizoma Tupistrae Chinensis saponin 10kg.Get mixture, add starch, granulate, granulate, encapsulated, contain saponin 10mg in every capsules.(used adjuvant can be one or more in capsules such as microcrystalline Cellulose, modified starch, ethyl cellulose, methylcellulose, hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, the sodium carboxymethyl cellulose adjuvant commonly used).
The preparation of embodiment 5 medicine capsules of the present invention
Get Rhizoma Tupistrae Chinensis aqueous extract 8kg, Rhizoma Tupistrae Chinensis lipoclastic 5kg, Rhizoma Tupistrae Chinensis saponin 10kg.Get mixture, add starch, granulate, granulate, encapsulated, contain saponin 8mg in every capsules.(used adjuvant can be one or more in capsules such as microcrystalline Cellulose, modified starch, ethyl cellulose, methylcellulose, hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, the sodium carboxymethyl cellulose adjuvant commonly used).
The preparation of embodiment 6 medicinal tablets of the present invention
Rhizoma Tupistrae Chinensis aqueous extract 10kg, the Rhizoma Tupistrae Chinensis lipoclastic 10kg, the Rhizoma Tupistrae Chinensis saponin 10kg that get embodiment 1 preparation mix.Get mixture, add starch and 1% magnesium stearate, tabletting makes every to contain saponin 10mg.(used adjuvant can be one or more in tablets such as microcrystalline Cellulose, modified starch, ethyl cellulose, methylcellulose, hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, the sodium carboxymethyl cellulose adjuvant commonly used).
The preparation of embodiment 7 medicinal tablets of the present invention
The Rhizoma Tupistrae Chinensis saponin 10kg that gets embodiment 1 preparation mixes.Get mixture, add starch and 1% magnesium stearate, tabletting makes every to contain saponin 5mg.(used adjuvant can be one or more in tablets such as microcrystalline Cellulose, modified starch, ethyl cellulose, methylcellulose, hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, the sodium carboxymethyl cellulose adjuvant commonly used).
Embodiment 8 medicine HPLC controls of the present invention
The HPLC test condition
The Waters-2690 pump, detector; Chromatographic column YMC-Pack ODS-AQ, s-6um, 120,250mm * 4.6mmI.D.; Mobile phase is that acetonitrile-buffering is to gradient (K
2HPO
4/ KH
2PO
4, concentration is 1mmol/L, pH value is 5.83), acetonitrile and K
2HPO
4/ KH
2PO
4The volume ratio initial value be 20: 80,10min is 22: 78,15min is 25: 75,20min is 28: 72,34min is 34: 66,44min is 43: 57,48min is 80: 20,52min is 85: 15,60min is 20: 80; Flow velocity is 1.0ml/min; Column temperature is 40 ℃; The detection wavelength is 203nm.
The results are shown in Figure 1.
Below prove beneficial effect of the present invention by pharmacodynamics test.
The screening test of test example 1 medicine Tupistra chinensis Bak. active site of the present invention
1, medicine
1.1 the preparation of Tupistra chinensis Bak. water extract
Get 200 gram Tupistra chinensis Bak. powder, add suitable quantity of water extract 55.4g extractum, as water extract group medicine.
1.2 the preparation of Tupistra chinensis Bak. methanolic extract
Get 150 gram Tupistra chinensis Bak. powder arts, add an amount of methanol extraction and obtain 70g Tupistra chinensis Bak. powder, as methanolic extract group medicine.
1.3 reagent dimethylbenzene (AR), Jinan Jiaozuo City chemical industry three factories.
2, animal Kunming mouse, body weight (22 ± 2) g is provided by SanXia University's Experimental Animal Center.The quality certification number: 0035537
3, instrument
-60 ℃ of horizontal cryostates, Great Wall, Shenyang medical apparatus and instruments factory; Electronic analytical balance, METTLER TOLEDO company; The FD-1 freezer dryer, Beijing rich doctor health technology company; Bacillus Calmette-Gu, Shanghai syringe factory.
4, method and result
30 of mices, ♂ is divided into 5 groups at random.Each organizes the same group of dosage ig administration.Once a day, continuous 7 days, 1h after the last administration, each group mouse right ear upper and lower faces is smeared dimethylbenzene 20 μ l respectively once cause inflammation, mice is put to death in left ear contrast (not being coated with) behind the 20min, cut ears, sweep away auricle with the card punch of 6mm diameter, immediately at scales/electronic balance weighing and record.Deduct left ear weight as the swelling degree with auris dextra weight, calculate inhibitory rate of intumesce.The results are shown in Table 1.
Inhibitory rate of intumesce=(negative control group difference-administration group difference)/negative control group difference * 100%
The comparison of table 1 Tupistra chinensis Bak. extract antiinflammatory action (X ± S)
Group dosage (gkg
1) number of animals swelling degree suppression ratio
Normal saline group 10 1.60
Methanolic extract group 15 10 1.42 ± 0.35
*11.25
Water extract group 15 10 1.53 ± 0.27 4.38
Annotate:
*Compare with the NS group P＜0.05.
5 discuss
Experimental result shows that Tupistra chinensis Bak. has certain antiinflammatory action.Wherein, the antiinflammatory action of methanolic extract is the strongest, and the efficient height of methanol extraction.For this reason, select methanolic extract to do the pharmacodynamic experiment of Tupistra chinensis Bak..
Test example 2 Tupistra chinensis Bak. antiinflammatory action experimentatioies
1, medicine methanol (AR); NH
4Cl (AR), Henan Jiaozuo City chemical industry three factories; Dimethylbenzene (AR), Henan Jiaozuo City chemical industry three factories.
2, instrument Rotary Evaporators, Wuxi City star sea king's biochemical equipment company limited; Electronic analytical balance, METTLERTOLEDO company.
3, animal Kunming mouse, body weight (22 ± 2) g is provided by SanXia University's Experimental Animal Center.
4, method and result
72 of mices, ♂ is divided into 6 groups at random, and each group is pressed document [1] record dosage ig administration.Positive controls ig ibuprofen 0.15g.kg
-1, negative control group is given with volume NS.Once a day, continuous 7 days, 1h after the last administration, each group mouse right ear upper and lower faces is smeared dimethylbenzene 20 μ l respectively once cause inflammation, mice is put to death in left ear contrast (not being coated with) behind the 15min, cut ears, sweep away auricle with the card punch of 6mm diameter, immediately at scales/electronic balance weighing and record.Deduct left ear weight as the swelling degree with auris dextra weight, calculate inhibitory rate of intumesce.The results are shown in Table 2.
Inhibitory rate of intumesce=(negative control group difference-administration group difference)/negative control group difference * 100%
Table 2 Tupistra chinensis Bak. each several part extract antiinflammatory action experimentation
Group dosage (mgkg
-1) swelling degree suppression ratio (%)
Normal saline group 4.87 ± 0.66
Ibuprofen group 90 4.70 ± 0.21
*3.49
Saponin high dose group 148.3 4.67 ± 0.32
*4.11
The dosage group 74.2 3.43 ± 0.14 in the saponin
*29.57
Liposoluble constituent group 7.2 3.63 ± 0.37
*25.46
Water soluble ingredient group 110.62 3.93 ± 0.58 19.30
Annotate:
*Compare with the NS group, P＜0.05,
*Compare with the ibuprofen group P＜0.01.
5, discuss
This shows that the antiinflammatory action of Tupistra chinensis Bak. is the coefficient result of each constituents, and wherein the antiinflammatory action of n-butyl alcohol part (being saponin) is the strongest, selecting n-butyl alcohol thus for use partly is that total saponins partly carries out following antiinflammatory experiment.
Test example 3 Tupistra chinensis Bak. water soluble ingredients, liposoluble constituent, the screening of saponin part proportioning
72 of mices, ♂ is divided into 6 groups, gastric infusion at random.Positive controls is irritated stomach aspirin 150mg.kg
-1, negative control group is given with the volume normal saline.Once a day, continuous 7 days, 1h after the last administration, each group mouse right ear upper and lower faces is smeared dimethylbenzene 20 μ l respectively once cause inflammation, mice is put to death in left ear contrast (not being coated with) behind the 20min, cut ears, sweep away auricle with the card punch of 6mm diameter, immediately at scales/electronic balance weighing and record.Deduct left ear weight as the swelling degree with auris dextra weight, calculate inhibitory rate of intumesce.The results are shown in Table 3.
Inhibitory rate of intumesce=(negative control group difference-administration group difference)/negative control group difference * 100%
Table 3 Tupistra chinensis Bak. each several part extract antiinflammatory action proportioning screening experiment research X ± S
Group dosage (mgkg
-1) swelling degree (mg) suppression ratio (%)
Normal saline group 6.72 ± 0.74
Aspirin group 150 4.93 ± 0.4** 26.6
The hydrotrope, lipoclastic, 50: 50: 5 5.08 ± 0.57** 24.4 of soap
Glycosides (10: 10: 1)
The hydrotrope, lipoclastic, 150: 50: 5 5.14 ± 0.54** 23.5 of soap
Glycosides (30: 10: 1)
The hydrotrope, lipoclastic, 50: 50: 15 3.96 ± 0.43** 41.1 of soap
Glycosides (10: 10: 3)
Saponin (10) 50 5.29 ± 0.48** 21.3
Annotate: * * and normal saline group compare, P＜0.01
Discuss
This shows that the antiinflammatory action of Tupistra chinensis Bak. mainly is liposoluble constituent, the synergistic result of saponin part, the water soluble ingredient influence is less, and the best proportioning of three constituents is the hydrotrope, lipoclastic, saponin (10: 10: 3).
The experimentation of test example 4 Rhizoma Tupistrae Chinensis saponins treatment pharyngitis
1, material
1.1 animal
15 of large ear rabbits, male and female half and half, body weight 2.02 ± 0.17kg raises with full-valence pellet feed, and regularly clean every day, provided by SanXia University's Experimental Animal Center.The quality certification number: 0035537.
1.2 medicine is by the saponin of embodiment 1 preparation
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water with 1: 1 mixing, are got brown extractum with chloroform extraction earlier and preserve in refrigerator, and the aqueous solution after the extraction is dry must yellow extractum standby in the freezing preservation of refrigerator with water saturated n-butanol extraction.
1.3 reagent ammonia, concentration 25%-27%; Herba Pileae Scriptae Tabellae, Jiangzhong Pharmaceutical Factory, Jiangxi Prov.
1.4 instrument Daphne Larynol aerosol apparatus, Hong Kong Mei Jiakang medicines and health protection company.
2, method
2.1 the foundation of animal model
15 large ear rabbits are divided into model control group, Tupistra chinensis Bak. high dose group, Tupistra chinensis Bak. low dose group, Herba Pileae Scriptae group, normal control group at random, every group each 5.Model control group: the 1-3 days mornings, afternoon respectively spray its pharyngeal 1 time with 15% ammonia, lifts with aerosol apparatus spray 3 at every turn, and 4-7 days spray distilled water, blood smear is got in docking in the 8th day.Tupistra chinensis Bak. height, middle dosage group: pharyngeal spray ammonia is with the model matched group, 4-7 days with laryngeal spray to large ear rabbit spray Rhizoma Tupistrae Chinensis saponin suspension (being made into distilled water) according to people and the conversion of large ear rabbit body surface coefficient, docked in the 8th day and to get blood smear.The Herba Pileae Scriptae group: pharyngeal spray ammonia is with the model matched group, 4-7 days with laryngeal spray to large ear rabbit spray Herba Pileae Scriptae suspension (being made into distilled water) according to people and the conversion of large ear rabbit body surface coefficient, docked in the 8th day and to get blood smear.The normal control group: spray equivalent distilled water, blood smear is got in docking in the 4th day.After each treated animal is got blood smear, take off pharyngeal mucosa immediately behind the femoral artery sacrificed by exsanguination animal and undertissue makes sections observation.
2.2 observation index
2.2.1 objective indicator
Large ear rabbit pharyngeal mucosa and undertissue's section thereof are made pathology morphology and Ultrastructural observation.Docking was got blood smear and is measured routine blood test before each treated animal was put to death.Observe the pharyngeal situation of animal every day from modeling beginning in the 2nd day, comprise mucosa form, color and luster etc., put to death animal, and got pharyngeal sticking to mould and undertissue thereof and make sections observation in the 8th day.
2.2.2 auxiliary characteristics experiment each treated animal mode of appearance of observed and recorded every day is observed the pharyngeal situation of animal every day 1 time, comprises mucosa form, color and luster etc.
2.3 detection method
Test each treated animal administration on the 8th and finish the post-tensioning ridge and put to death, get pharyngeal mucosa and submucous tissue immediately and be divided into two parts and carry out pathomorphology and Ultrastructural observation respectively.Take out after observing the fixation of tissue 48h of pathomorphology, be cut to 2mm thickness, sheet is observed, is taken the photograph in dehydration routinely, transparent, paraffin embedding, section, HE dyeing under optical microscope.Observe Ultrastructural tissue and fix through 2.5% glutaraldehyde, the propionic aldehyde dehydration, embedding, ultrathin section, electron staining places and observes, takes the photograph sheet under the H-500 transmission electron microscope.
2.4 statistical method
Adopt the SPSS11.0 statistical software, the result represents that with x ± S group difference relatively adopts the t check.
3, result
3.1 general state
Model control group is from modeling the 2nd day, and most of large ear rabbit engenders to disturb grabs oral area, drinking-water and amount is many, pharyngeal congestion is symptom and Signs such as cerise, swelling frequently, and symptom was more obvious in the 3rd day; Above-mentioned situation does not then appear in the normal control group; The Tupistra chinensis Bak. high dose group, the Herba Pileae Scriptae treatment is after 4 days, and it is normal that above-mentioned symptom, sign are recovered substantially; It is poor slightly that low dose group is recovered; The above-mentioned symptom of model control group, sign then still exist.
3.2 the routine blood test index changes
The results are shown in Table 4
Each animal groups routine blood test variation of table 4 (x ± S)
The number of animals total white blood cells
Group (only) (10
9/ L)
Normal control group 5 5.12 ± 1.21
*
Model control group 5 9.46 ± 1.34
High dose group 5 6.56 ± 1.12
* ※ ※
Low dose group 5 8.23 ± 1.14
* ※
Herba Pileae Scriptae group 5 7.36 ± 1.13
*
Annotate: compare with model control group,
*P＜0.01; Compare with the Herba Pileae Scriptae group,
※P＜0.05
※ ※P＜0.01
3.3 the pharyngeal pathology change
The normal control group: large ear rabbit pharyngeal mucous epithelium comes in every shape in different parts.This research large ear rabbit mucous epithelium is a stratified squamous epithelium, and mucous epithelium for lamina propria, is made up of dense connective tissue down, dispersive lymphatic nodule and diffuse lymphoid tissue are arranged in the lamina propria, also have more mucous gland, lamina propria lower floor is the flesh layer, elastic fiber is more, forms the elastic fiber film.Model control group: the outer keratinization of the mucous epithelium of pharynx tissue, part comes off, the multiple layer of mucous epithelium is hypertrophy obviously, tela submucosa is very thin does not have obvious boundary with lamina propria, little vasodilation hyperemia in the lamina propria, edema, the necrosis of part mucous gland epithelial cell, downright bad mucous gland body forms cavity or forms structureless mucus netted; A large amount of inflammatory cell infiltrations is arranged between lamina propria zone and mucous gland, and lamina propria lower floor is the flesh layer, flesh layer muscle fiber irregular arrangement.The Herba Pileae Scriptae group: pharynx tissue adherence stratified squamous epithelium, last outer cortex keratinization, part comes off, mucous epithelium for lamina propria, is made up of dense connective tissue down, and many mucous glands are arranged in the lamina propria, the necrosis of mucous gland epithelial cell seldom has slight inflammatory cell infiltration in the lamina propria and between mucous gland.High dose group: pharynx tissue adherence stratified squamous epithelium, last outer cortex keratinization, the stratified squamous epithelium hypertrophy is not obvious, and more mucous gland is arranged in the lamina propria, and slight inflammatory cell infiltration is arranged between lamina propria and mucous gland, shows that the pharynx tissue has greatly improved.Low dose group: the mucosa stratified squamous epithelium of pharynx tissue, do not thicken, lamina propria under the mucosa, there are many mucous glands to distribute therein, the inflammatory cell infiltration that moderate is arranged between lamina propria and mucous gland, lamina propria lower floor is the flesh layer, muscle fiber is arranged not very rule, its pathological changes and normal control group are poorer, but more also make moderate progress with model group.
3.4 pharyngeal Change of Ultrastructure
The normal control group: the nucleus of cell (N) nuclear membrane is double-deck under the large ear rabbit pharyngeal mucosa, and Nuclear pore (NP) arranged, be dispersed in more ribosome (RI) in the kytoplasm, ribosome is attached to constituting rough surface endoplasmic reticulum (RER) on the membrana sacciformis, some regional accidental ribosome membrana sacciformis bright finish type endoplasmic reticulum (SER) that has, mitochondrion (MI) is shaped as circle or ellipse, surround by duplicature, the inner membrance invaginate constitutes ridge, the basophilic granule that engrain is arranged in the substrate between ridge, the various lysosome (LY) of content form that monolayer surrounds.Model control group: confluent monolayer cells pyknosis under the pharyngeal mucosa, ultrastructure is destroyed, and organelle disintegrates, has the nuclei dyeing chromaticness to be irregular netted in the middle of it; The obvious degeneration of mitochondrion, arrangement disorder, swelling, ridge reduces and breaks, the formation myelin figure that has, minority forms the pile tube ridge, and the substrate electron density that has reduces, and makes substrate be mottled, cavity change; The kytoplasm rough endoplasmic reticulum is significantly expanded.The Herba Pileae Scriptae group: the slight degeneration of the mitochondrion of cell under the pharyngeal mucosa, most of mitochondrial crista do not reduce and shorten, and have only the minority ridge to reduce and shorten, and the substrate electron density reduces, and other organelles change not obvious.High dose group: the nucleus of cell (N) nuclear membrane is double-deck under the pharyngeal mucosa, nuclear chromatin is evenly distributed, the structure of mitochondria of most of cell changes not obvious, mostly be circular or oval, surrounded by duplicature, the inner membrance invaginate constitutes ridge, but the slight degeneration of minority mitochondrion is also arranged, ridge reduces or shortens, and the ultrastructure that shows cell makes moderate progress and recovers.
Above-mentioned test explanation, medicine Rhizoma Tupistrae Chinensis saponin of the present invention has the effect of treatment pharyngitis, and positive drug is a Herba Pileae Scriptae Tabellae, illustrates that medicine of the present invention has the effect identical with positive drug, all has oral, as to suck treatment pharyngitis effect.
Test example 5 medicine Rhizoma Tupistrae Chinensis saponin bacteriostatic experiments of the present invention
1, material
1.1 culture medium
Broth bouillon and nutrient agar all dispose by operation instruction.
1.2 antibacterial
1.2.1 strain
Staphylococcus aureus (ATCC25923), colon bacillus (ATCC25922), the false monospore (ATCC27853) of Aerugo is provided by Medical College of Sanxia University's microbiology and immunology teaching and research room; Beta hemolytic streptococcus, klebsiella pneumoniae, staphylococcus epidermidis is provided by Yichang No. 2 People's Hospital.
1.2.2 bacteria suspension preparation
Before the experiment various antibacterials are got 1 oese (or inoculating loop) respectively and be inoculated in the 10ml broth bouillon, cultivate 18h in 37 ℃.With this bacterial suspension inoculation in Nutrient agar, cultivate 24 h after the single colony inoculation of picking in the 5ml broth bouillon, be diluted to 10 with NS from stock solution after cultivating 18h
-7Totally 7 dilution factors, obtaining concentration with the flat-plate bacterial colony numeration then is 105CFUml
-1Bacteria suspension standby.
1.3 antibacterials: by the saponin of embodiment 1 preparation.
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water were fully rotated mixing with 1: 1, use earlier chloroform extraction,, the n-butyl alcohol extract concentrate drying is got total saponins (the molish reaction is positive), be and be subjected to reagent the water saturated n-butanol extraction of remaining aqueous solution after the extraction.Get Rhizoma Tupistrae Chinensis saponin 20g and add the stock solution that the 20ml distilled water diluting becomes 1000mg/ml, standby with the cold preservation of Pasteur's moral method.
2, method and result (test tube doubling dilution)
To 60 in the band plug sterilization test tube of 1.0ml meat soup be housed, press the 1-10 numbering, divide 6 groups and be arranged in test tube rack, by the sterile working, in first row's test tube, add Tupistra chinensis Bak. stock solution 1.0ml respectively, take out 1.0ml behind the mixing and put into the 2nd pipe, get 1.0ml from the 2nd pipe behind the same mixing and put into the 3rd pipe, method is diluted to the 9th pipe by pipe according to this.The 10th pipe does not add medicine manages in contrast, and each pipe all adds concentration and is about 5 * 10
5CFUml
-1Each cause of disease indicator bacteria bacterium liquid 0.1ml, put into 37 ℃ of incubators behind the mixing and cultivate 24h.Above-mentioned test tube culture fluid is got 100 μ l respectively coat nutrient agar panel, parallel 2 wares of doing of every pipe, observed result behind 37 ℃ of cultivation 18h.Repeat 2 times, asepsis growth represents that this pipe dose promptly is the minimum inhibitory concentration (MIC) that is subjected to reagent.The results are shown in Table 5.
The external bacteriostatic activity of table 5 Tupistra chinensis Bak.
Strain dilution factor minimum inhibitory concentration (gml
-1)
Staphylococcus aureus 64 0.063
Staphylococcus epidermidis 64 0.063
The false monospore 64 0.063 of Aerugo
Beta hemolytic streptococcus 8 0.25
Escherichia coli 8 0.25
Klebsiella pneumoniae 8 0.25
The result shows that Tupistra chinensis Bak. has the good in-vitro bacteriostasis to bottleneck throat common pathogen such as staphylococcus aureus, beta hemolytic streptococcus etc.
The 6 medicine Rhizoma Tupistrae Chinensis saponin phlegm-dispelling functions experiments of the present invention of test example
1, material
1.1 the preparation of Tupistra chinensis Bak. extract
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water were fully rotated mixing with 1: 1, use earlier chloroform extraction,, the n-butyl alcohol extract concentrate drying is got total saponins (the molish reaction is positive), be and be subjected to reagent the water saturated n-butanol extraction of remaining aqueous solution after the extraction.
1.2 medicine methanol (AR); NH
4Cl (AR), Henan Jiaozuo City chemical industry three factories; Phenol red, recovery fine chemistry industry institute; NaHCO
3The north of the Changjiang River, Wuhan City chemical reagent factory.
1.3 the instrument Bacillus Calmette-Gu, south, the west of a city, Jintan City township Yao Zhusheqichang makes; UV-754 type ultraviolet-uisible spectrophotometer, Shanghai the 3rd analytical tool factory.
1.4 the animal Kunming mouse, body weight (22 ± 2) g is provided by SanXia University's Experimental Animal Center.The quality certification number: 0035537
2 methods and result
50 of mices, ♀ ♂ half and half is divided into 5 groups at random.(dosage is pressed LD to each group by various dose
501/10 calculate, 2,3 groups more last group is successively decreased half) the ig administration, the administration volume is 10mlkg
-1Body weight, positive controls is given 0.15gkg
-1NH
4Cl solution, negative control group is given the NS with volume, once a day, gives 7 times altogether, hungry 16h before the art time administration, 30min after the administration, lumbar injection 5% phenol red NS solution 0.5ml behind the injection 30min, puts to death animal, and back of the body position is fixing, separates trachea, uses 5%NaHCO
3The solution flushing, each 1.0ml washes 3 times altogether, merges flushing liquor, measures the OD value at 546nm wavelength place, compares with matched group.The results are shown in Table 6.
Table 6 Tupistra chinensis Bak. extract to the influence of the disconnected phenol red excretion of mice trachea (x ± S, n=10)
Group dosage (mgkg
-1) the OD value
Normal saline 2.49 ± 0.04
Rhizoma Tupistrae Chinensis saponin 2.45 2.72 ± 0.02
* *
Rhizoma Tupistrae Chinensis saponin 1.23 2.68 ± 0.01
* *
Rhizoma Tupistrae Chinensis saponin 0.62 2.67 ± 0.01
* *
Ammonium chloride 0.15 2.57 ± 0.03
*
**P＜0.05???
***P＜0.001?vs?NS
The result shows that the 0D value of the phenol red excretion amount of the high, medium and low dosage group of Tupistra chinensis Bak. trachea section for high, has significant differences (P＜0.001) than the NS group, and NH
4The Cl group more only has significant difference (P＜0.05) than the NS group.
Test example 7 medicine Rhizoma Tupistrae Chinensis saponins of the present invention are to the influence of delayed hypersensitivity
1, material
1.1 the preparation of Tupistra chinensis Bak. extract: the method preparation of pressing embodiment 1
48 ℃ of dried overnight of Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water were fully rotated mixing with 1: 1, use earlier chloroform extraction,, the n-butyl alcohol extract concentrate drying is got total saponins (the molish reaction is positive), be and be subjected to reagent the water saturated n-butanol extraction of remaining aqueous solution after the extraction.
1.2 medicine DNCB (AR), Shanghai Pujiang chemical industry company limited; Acetone (AR), the north of the Changjiang River, Wuhan City chemical reagent factory.
1.3 the instrument microsyringe, Shanghai Medicine Laser Instrument Plant.
1.4 the animal Kunming mouse, body weight (22 ± 2) g is provided by SanXia University's Experimental Animal Center.The quality certification number: 0035537
2, method
2.1 influence to DNCB induced mice DTH
40 of mices, ♀ ♂ half and half is divided into 4 groups at random.Every mouse web portion cropping, the about 2cm of area * 2cm size, 50 μ l evenly spread upon cropping district sensitization with the 1.25%DNCB acetone soln, and (dosage is pressed LD
501/10 calculate, 2,3 groups more last group is successively decreased half) ig administration and sensitization every day 1 time, continuous 8 days, 1.25%DNCB acetone soln 20 μ l evenly were applied in the mouse right ear two sides in the 8th day after the sensitization and attack, left ear is coated with the contrast of equal volume acetone, and the cervical vertebra dislocation is put to death behind the 24h, sweep away left and right sides auricle with diameter 6mm card punch, on electronic balance, weigh immediately, deduct left ear weight as the swelling degree, calculate inhibitory rate of intumesce with auris dextra weight.The results are shown in Table 7.
Inhibitory rate of intumesce=(negative control group difference-administration group difference)/negative control group difference * 100%
3, result
Table 7 Tupistra chinensis Bak. extract to the influence of DNCB induced mice DTH (x ± S, n=10)
Group dosage (mgkg
-1) swelling degree suppression ratio (%)
Normal saline 8.82 ± 0.55
Rhizoma Tupistrae Chinensis saponin 2.45 5.91 ± 1.39 32.99
Rhizoma Tupistrae Chinensis saponin 1.23 6.55 ± 0.90 25.74
Rhizoma Tupistrae Chinensis saponin 0.62 8.20 ± 0.74 7.03
Annotate: compare P＞0.05 with the normal saline group
The result shows, continuous 8 days of Rhizoma Tupistrae Chinensis saponin ig administration does not make significant difference (P＞0.05) to the inductive mouse DTH of DNCB, illustrates that Rhizoma Tupistrae Chinensis saponin has certain anti-allergic effects, but a little less than the effect.
Test example 8 medicine Rhizoma Tupistrae Chinensis saponin acute toxicity testings of the present invention
1, material
1.1 48 ℃ of dried overnight of medicament preparation Tupistra chinensis Bak. rhizome, cracker for medicine is pulverized the back and is crossed 60 mesh sieves, is solvent with methanol, adopts multi-functional pressure percolation jar in normal pressure, 65 ℃ of extractions down, concentrate (temperature: 48 ℃) with Rotary Evaporators behind the extracting liquid filtering, the dry dark-brown extractum that gets.This extractum and distilled water were fully rotated mixing with 1: 1, use earlier chloroform extraction,, the n-butyl alcohol extract concentrate drying is got total saponins (the molish reaction is positive), be and be subjected to reagent the water saturated n-butanol extraction of remaining aqueous solution after the extraction.
1.2 the animal Kunming mouse, body weight 18-22g, male and female half and half, pregnant Mus need not, dosage 0.2mL/10g.
1.3 the instrument Bacillus Calmette-Gu, Shanghai syringe factory.
2, method and result
This test method adopts Sun Shi improvement Guan Shi method to survey median lethal dose(LD 50) LD
50, be divided into prerun experiment and formal experiment.
2.1 prerun experiment
The prerun experiment: 10 of white mice, with 100%, 200%, 300% Tupistra chinensis Bak. experimentizes.Result: 100%:4 is only all normal.200%:3 only tests, and is all normal.300%:3 only tests, dead 2.(observing after 24 hours).Perfusion 200%: poured into behind 100% medicinal liquid 4 hours, 100% medicinal liquid of same dosage is poured in fasting again after 5 hours.Perfusion 300%: with 100% medicinal liquid perfusion 3 times, perfusion in per 5 hours is once poured into fasting in preceding 1 hour.
Observe after one week, 200% is all dead, 300% only 1 survival, and other gets 15 white mice, joins 17.5ml100% Tupistra chinensis Bak. liquid, does 100%, 200% experiment.Experimental result: 100% is all normal, and 200% is all dead.(5 every group)
Pre-test result: tentatively measure the dosage range of mortality rate at 0%-100%, experimental result shows.Mortality rate is 100% at 0% Cmax, and mortality rate is 200% at 100% Cmin.
2.2 formal experiment
50 of mices are divided into 5 groups at random, weigh and labelling, and concentration group common ratio is 0.7 between group, behind the gastric infusion, observes each death toll after 24 hours.
2.2.1 calculate every group of dose
V=10 (group number) * 20 (every treated animal body weight) * 0.2ml/10g=4ml is for avoiding waste every assembly 5ml medicinal liquid.
2.2.2 calculate mother solution volume and concentration
V mother solution=5/ (1-0.7)=17ml C concentration=200% (liquor strength during maximum lethal dose)
2.2.3 join solution-low ratio serial dilution
Mother solution: get the 34g adding distil water to 17ml, promptly get 200% medicinal liquid.
Drawing 5ml from the 17ml mother solution places dry No. 1 medicine bottle (concentration 200%) for first group of usefulness; In remaining 12ml mother solution (content of dispersion 24.0g), add the 5ml distilled water, draw 5cm and be put in No. 2 bottles (concentration 142.2%), for second group of use to 17ml; In remaining 12ml medicinal liquid (content of dispersion 16.9g), add the 5ml distilled water, draw 5ml and be put in kingpin (concentration 49.6%), for the 5th group of use to 17ml.
2.2.4 calculating median lethal dose(LD 50)
LD50＝lg-1[Xm-I*(∑P-0.5)]
Wherein Xm is the logarithm value of maximal dose, and I is the logarithm value of two adjacent groups dose ratio, and P is each group death toll.
2.2.5 experiment relative recording
Table 8 Tupistra chinensis Bak. acute toxicity testing (n=10)
Dosage (mgkg
-1) the death toll mortality rate
159.9????????????4????????????0.4
113.6????????????2????????????0.2
79.6?????????????1????????????0.1
56.4?????????????0????????????0
39.7?????????????0????????????0
Calculate LD by formula
50=37.19gkg
-1, be equivalent to crude drug 148.7gkg
-1, promptly be equivalent to the day for human beings 2974 times with dosage, hence one can see that, and the toxicity of Tupistra chinensis Bak. is very little.
Vertical the above, Rhizoma Tupistrae Chinensis aqueous extract of the present invention, Rhizoma Tupistrae Chinensis lipoclastic, Rhizoma Tupistrae Chinensis saponin three compositions are all effective, and use at the extract compatibility, have brought into play the effect of Synergistic, and safe, controllability is strong, and is stable, provides a kind of new selection for clinical.
List of references is as follows:
1. the new medical college in Jiangsu is compiled Chinese medicine dictionary, the first volume, Shanghai: Shanghai science tech publishing house, 1985,10.pp?907
2. Hubei Province health bureau of revolutionary committee compiles, Hubei Chinese herbal medicine will (one), Wuhan: People's Health Publisher, Hubei, 1978,6.pp?146
3. the Lanzhou health subdepartment of military area logistics department is compiled, the sweet peaceful blue or green Chinese herbal medicine choosing in Shan, 1971.pp?236
4. national Chinese herbal medicine compilation group is compiled, national Chinese herbal medicine compilation, the first volume.Beijing: People's Health Publisher, 1975,9.pp?365
5. scholar's Qu brightness, Tujia's medicine Tupistra chinensis Bak. collutory clinical practice treatment pharyngolaryngitis 45 examples [J], Chinese national folk medical magazine, 1999, (3): 140
6. yellow beautiful, Liao Quanbin, Zou Kun, Ruan Hao, Nie Dandan, Hu Chaolin, Hu Yanliang.The assay of steroid sapogenin in the Tupistra chinensis Bak..SanXia University's journal (natural science edition) 2003,25,6,562-564 (instructing undergraduate's paper)
7.Pan，W.B.；Chang，F.R.；Wu，Y.C.J.Nat.Prod.2000，63，861-863.
8.Pan，W，B.；Chang，F.R.；Wu，Y.C.Chem.Pharm.Bull.2000，48，1350-1353.
9.Pan，W.B.；Chang，F.R.；Wei，L.M.；Wu，Y.C.J.Nat.Prod?2003，65，232-239.
Claims (11)
1, Tupistra chinensis Bak. Tupistra Chinensis Bak. extract, it is characterized in that: it contains the component of following weight proportion:
0～10 part of Rhizoma Tupistrae Chinensis aqueous extract, 0～10 part of Rhizoma Tupistrae Chinensis lipoclastic, 1～10 part of Rhizoma Tupistrae Chinensis saponin.
2, Tupistra chinensis Bak. extract according to claim 1, it is characterized in that: it contains the component of following weight proportion:
10 parts of Rhizoma Tupistrae Chinensis aqueous extracts, 10 parts of Rhizoma Tupistrae Chinensis lipoclastics, 3 parts of Rhizoma Tupistrae Chinensis saponins.
3, Tupistra chinensis Bak. extract according to claim 1 and 2 is characterized in that: described Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, saponin are by following method preparation:
With Liliaceae Herba Convallariae family Tupistra chinensis Bak. platymiscium Tupistra chinensis Bak. Tupistra Chinensis Bak. rhizome is raw material, is solvent with methanol, extracts the dry extractum that gets, and is methanolic extract; This extract and distilled water are with 1: 1 mixing of volume ratio, earlier with chloroform extraction, the dry Rhizoma Tupistrae Chinensis lipoclastic that gets; Aqueous solution after the extraction is with water saturated n-butanol extraction, dry Rhizoma Tupistrae Chinensis saponin; The dry Rhizoma Tupistrae Chinensis aqueous extract that gets of remaining aqueous solution.
4, a kind of pharmaceutical composition, it is to be mixed into active component by one of Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, saponin or its, adds acceptable accessories or complementary composition, the preparation that is prepared from.
5, pharmaceutical composition according to claim 4 is characterized in that: the HPLC finger printing of this pharmaceutical composition is made up of 5 characteristic peaks as shown in Figure 1,
Wherein chromatographic condition is: chromatographic column YMC-Pack ODS-AQ, s-6um, 120,250mm * 4.6mmI.D.; Mobile phase is acetonitrile-K
2HPO
4/ KH
2PO
4, gradient buffer, wherein K
2HPO
4/ KH
2PO
4Concentration is 1mmol/L, and pH value is 5.83, acetonitrile and K
2HPO
4/ KH
2PO
4The volume ratio initial value be 20: 80,10min is 22: 78,15min is 25: 75,20min is 28: 72,34min is 34: 66,44min is 43: 57,48min is 80: 20,52min is 85: 15,60min is 20: 80; Flow velocity is 1.0ml/min; Column temperature is 40 ℃; The detection wavelength is 203nm.
6, pharmaceutical composition according to claim 5 is characterized in that: the retention time RT value of described 5 characteristic peaks is respectively: No. 1 peak: 23.2min ± 0.86; No. 2 peak: 24.6min ± 0.90; No. 3 peak: 27.2min ± 0.84; No. 4 peak: 30.4min ± 0.96; No. 5 peak: 33.8min ± 0.98.
7, a kind of method for preparing the described pharmaceutical composition of claim 4 comprises the steps:
The preparation of a, Tupistra chinensis Bak. extract: with Liliaceae Herba Convallariae family Tupistra chinensis Bak. platymiscium Tupistra chinensis Bak. TupistraChinensis Bak. rhizome is raw material, is solvent with methanol, extracts the dry extractum that gets, and is methanolic extract; This extract with and distilled water with 1: 1 mixing of volume ratio, earlier with chloroform extraction, dry Rhizoma Tupistrae Chinensis lipoclastic; Aqueous solution after the extraction is with water saturated n-butanol extraction, dry Rhizoma Tupistrae Chinensis saponin; The dry Rhizoma Tupistrae Chinensis aqueous extract that gets of remaining aqueous solution;
B, Rhizoma Tupistrae Chinensis aqueous extract, lipoclastic, the saponin of getting a step preparation are active component, press column weight amount proportioning and add acceptable accessories or complementary composition, the preparation that is prepared from:
0～10 part of Rhizoma Tupistrae Chinensis aqueous extract, 0～10 part of Rhizoma Tupistrae Chinensis lipoclastic, 1～10 part of Rhizoma Tupistrae Chinensis saponin.
8, the purposes of the described pharmaceutical composition of claim 4 in the preparation anti-inflammatory drug.
9, purposes according to claim 8 is characterized in that: described medicine is the medicine of treatment pharyngitis.
10, purposes according to claim 9 is characterized in that: described medicine be suck, oral drugs, injectable drug.
11, the purposes of the described pharmaceutical composition of claim 4 in the preparation expelling phlegm drugs.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103520445A (en) * | 2013-10-28 | 2014-01-22 | 武汉大学 | Application of Tupistra chinensis in preparation of antitumor drugs |
| CN103623186A (en) * | 2013-12-06 | 2014-03-12 | 李凯 | Traditional Chinese medicine for treating pseudomembranous colitis |
| CN106509281A (en) * | 2016-12-21 | 2017-03-22 | 神农架林区中医医院 | campylandra chinensis sore-throat relieving tea and preparation method thereof |
| CN107361403A (en) * | 2017-08-07 | 2017-11-21 | 武汉纺织大学 | One kind contains Extracts of Tupistra chinensis additive in cigarette and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100348109C (en) * | 2004-02-27 | 2007-11-14 | 华南农业大学 | Method for extractiong microbicide and anti-staling agent from tupistra ker, and its application |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103520445A (en) * | 2013-10-28 | 2014-01-22 | 武汉大学 | Application of Tupistra chinensis in preparation of antitumor drugs |
| CN103623186A (en) * | 2013-12-06 | 2014-03-12 | 李凯 | Traditional Chinese medicine for treating pseudomembranous colitis |
| CN106509281A (en) * | 2016-12-21 | 2017-03-22 | 神农架林区中医医院 | campylandra chinensis sore-throat relieving tea and preparation method thereof |
| CN107361403A (en) * | 2017-08-07 | 2017-11-21 | 武汉纺织大学 | One kind contains Extracts of Tupistra chinensis additive in cigarette and preparation method thereof |
| CN107361403B (en) * | 2017-08-07 | 2018-09-18 | 武汉纺织大学 | A kind of cigarette of additive in cigarette and preparation method thereof containing Extracts of Tupistra chinensis |
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