CN104130338B - The preparation method and applications of Buick gracilis polysaccharide - Google Patents
The preparation method and applications of Buick gracilis polysaccharide Download PDFInfo
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- CN104130338B CN104130338B CN201410382303.4A CN201410382303A CN104130338B CN 104130338 B CN104130338 B CN 104130338B CN 201410382303 A CN201410382303 A CN 201410382303A CN 104130338 B CN104130338 B CN 104130338B
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Abstract
The present invention provides the preparation method of a kind of Buick gracilis polysaccharide, be Buick is joined medicinal powder alcohol extraction defat after, water extract-alcohol precipitation obtains Buick gracilis polysaccharide.The present invention also provides for the application in preparation antiinflammatory, analgesic of the Buick gracilis polysaccharide.Through test, Buick gracilis polysaccharide has good antiinflammatory and analgesic effect, it is possible to be used for preparing antiinflammatory, analgesic medicine.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to the preparation method and applications of Buick gracilis polysaccharide.
Background technology
Buick is joined, and (has another name called Siberia Erythronium japonicum, Altai Mountains pig for Liliaceae Erythronium japonicum platymiscium Xinjiang Erythronium japonicum
Tartar) dry meat bulb.Originate from In The North of Xinjiang (Tianshan Area to Altay, Fu Hai), be born in Schattenseite, leaf margin thick grass, woods
Under, under bushes and on sub-alpine meadow, height above sea level 1100-2500m.Also there is distribution in Siberia to Central Asia.June is at stem
Excavating when leaf is not yet withered, boiled water dries after slightly boiling.Property dry, micro-hardship, nourish, keep fit, strong body, how sick cure mainly body void, the acid of waist knee joint
Soft, unable, it is one of time-honored medicine food dual purpose plant of Xinjiang Kazak nationality.
Inflammation refer to that the damage that various pro-inflammatory cytokines cause occurred by the biological tissue with vascular system with defence
Reaction is main pathological process.Basic pathology is changed to go bad, ooze out and hypertrophy.Topical manifestations is red, swollen, hot, pain and function
Obstacle, general reaction has White blood cell etc. in heating, blood.Inflammation is found in the pathological process of multiple disease.Anti-inflammatory drug is
Refer to the class medicine that inflammatory reaction is had inhibitory action or conciliation effect.The antiinflammatory action of drugs, seeks safe and effective
Anti-inflammatory substance is the problem that numerous medicine scholar pays close attention to jointly.The antiinflammatory action of drugs, must first replicate inflammatory animal model,
Choose multiple model and carry out antiinflammatory screening, to reach drug effect is carried out the purpose of overall merit.
Mice caused by dimethylbenzene xylene ear swelling experimental model is typical acute inflammation model.Dominant mechanism is that dimethylbenzene is applied in
After Mice Auricle, cause some inflammatory mediator such as histamine, kassinin kinin and fibrinolysis to discharge, cause local vascular dilation, hair
Tubule permeability increases, and inflammatory cell infiltration causes ear's acute exudative inflammation edema.Comparative experiments group and matched group diformazan
The difference of ear swelling degree after benzene effect, to determine whether given the test agent has the effect of intervention acute inflammation (mice ear).
Mice granuloma induced by implantation of cotton pellets experimental model is typical subacute inflammation model.Pathologic Characteristics is with lymphocyte and list
Core macrophages infiltration is main, has the hypertrophy of the parenchymas such as obvious fibrous connective tissue, blood vessel and epithelial cell, i.e. meat
Bud.The difference of granulation tissue quality is generated, to determine whether given the test agent has intervention Asia by comparative experiments group and matched group
The effect of acute inflammation (mice granuloma induced by implantation of cotton pellets).
It is one of method verifying medicine analgesic effect that glacial acetic acid causes mouse writhing experimental model.Dominant mechanism is by necessarily
The chemical irritants of volume and concentration injects in mouse peritoneal, stimulates visceral layer and parietal peritoneum, cause larger area, longer time
Between inflammatory pain, cause mice that the behavior reactions, referred to as writhing such as abdominal part indent, trunk and hind leg are upheld, hips up occur
Reaction.In this reaction 15min after injection, frequency is higher, therefore determines as pain with the writhing number of times occurred in 15min after injection
Figureofmerit.By the difference of comparative experiments group Yu matched group writhing number of times, to determine whether given the test agent has analgesic effect.
Summary of the invention
The invention provides the preparation method of Buick gracilis polysaccharide and the application in preparing anti-inflammation analgesis medicament thereof.
The present invention provides the preparation method of a kind of Buick gracilis polysaccharide, be Buick is joined medicinal powder alcohol extraction defat after, water carries
Precipitate with ethanol obtains Buick gracilis polysaccharide.
Preferably, described alcohol extraction is ethanol.
Preferably, described method is: weighs Buick ginseng medicinal powder, adds 6-12 times of volume 70-100% ethanol, heat back
Stream extracts 1-5 time, extraction time 1-4h, collects medicinal residues and dries, and adds 6-30 times of volume of water 60-100 in medicinal residues after the drying
DEG C extracting 1-5 time, extraction time is 1-4h, merges water and carries filtrate, drying under reduced pressure to the most thick, 2-6 times of volume of addition anhydrous
Ethanol, cold preservation stands, has white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, and drying under reduced pressure is to weighing apparatus
Weight, to obtain final product.
Most preferably, described method is: weighs Buick ginseng medicinal powder, adds 12 times of volume 95% ethanol, be heated to reflux carrying
Take 5 times, extraction time 4h, collect medicinal residues and dry, medicinal residues after the drying add 30 times of volume of water 100 DEG C and extracts 5 times, extract
Time is 4h, merges water and carries filtrate, and drying under reduced pressure, to thick, adds the dehydrated alcohol of 6 times of volumes, and cold preservation stands, and has white
Polysaccharide separates out, and filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.
The present invention also provides for the Buick gracilis polysaccharide that should prepare with the aforedescribed process.
Third object of the present invention is to provide the application in preparation antiinflammatory, analgesic of the Buick gracilis polysaccharide.
Fourth object of the present invention is to provide a kind of antiinflammatory, analgesic, and its effective ingredient is Buick gracilis polysaccharide.
Through test, Buick gracilis polysaccharide has good effect to antiinflammatory, analgesia, it is possible to for preparing antiinflammatory, the medicine of analgesia class
Thing.
Detailed description of the invention
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiment
Method, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is certainly
Routine biochemistry reagent shop is commercially available.
1 material
1.1 medical material
Buick ginseng medical material picks up from Altay, Xinjiang, is accredited as liliaceous plant through pharmaceutical college of Xinjiang Medicine University
The bulb of Erythronium sibiricum (Fisch. et. Mey.) Kryl Xinjiang Erythronium japonicum.
1.2 animal
Kunming mice, body weight 18-22g, male, Xinjiang Medicine University's experimental animal center provide, normal raise 3d after
Start test.
1.3 reagent
Dehydrated alcohol, acetone, aspirin effervescent tablets (AstraZeneca pharmaceutical Co. Ltd, batch number 1208042), Ah
Amdinocillin capsule (Baiyunshan Pharmaceutical General Factory, batch number 4190047), dimethylbenzene, glacial acetic acid, ether.
1.4 equipment
Water-bath, Rotary Evaporators, vacuum drying oven, analytical balance, card punch.
Embodiment 1
The extraction preparation method of the Buick gracilis polysaccharide of the present invention is as follows:
Weigh Buick ginseng medicinal powder, add 6 times of volume 70% ethanol, heating and refluxing extraction 1 time, extraction time 1h, collect
Medicinal residues are placed in ventilation and dry, and dried medicinal residues are placed in round-bottomed flask, add 6 times of volume of water 60 DEG C and extract 1 time, during extraction
Between be 1h, merge water and carry filtrate, drying under reduced pressure to the most thick, adds the dehydrated alcohol of 2 times of volumes, puts in 4 DEG C of refrigerators and stands
24h, has white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, and 50 DEG C of drying under reduced pressure are to weight, i.e.
?.After measured, Buick gracilis polysaccharide yield: 3.12%.
Embodiment 2
The extraction preparation method of the Buick gracilis polysaccharide of the present invention is as follows:
Weigh Buick ginseng medicinal powder, add 7 times of volume 80% ethanol, heating and refluxing extraction 2 times, extraction time 2h, collect
Medicinal residues are placed in ventilation and dry, and dried medicinal residues are placed in round-bottomed flask, add 12 times of volume of water 70 DEG C and extract 2 times, extract
Time is 2h, merges water and carries filtrate, and drying under reduced pressure, to thick, add the dehydrated alcohol of 3 times of volumes, puts in 4 DEG C of refrigerators quiet
Putting 24h, have white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, 50 DEG C of drying under reduced pressure are to weight, i.e.
?.After measured, Buick gracilis polysaccharide yield: 6.24%.
Embodiment 3
The extraction preparation method of the Buick gracilis polysaccharide of the present invention is as follows:
Weigh Buick ginseng medicinal powder, add 8 times of volume 80% ethanol, heating and refluxing extraction 3 times, extraction time 3h, collect
Medicinal residues are placed in ventilation and dry, and dried medicinal residues are placed in round-bottomed flask, add 18 times of volume of water 80 DEG C and extract 3 times, extract
Time is 3h, merges water and carries filtrate, and drying under reduced pressure, to thick, add the dehydrated alcohol of 4 times of volumes, puts in 4 DEG C of refrigerators quiet
Putting 24h, have white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, 50 DEG C of drying under reduced pressure are to weight, i.e.
?.After measured, Buick gracilis polysaccharide yield: 9.38%.
Embodiment 4
The extraction preparation method of the Buick gracilis polysaccharide of the present invention is as follows:
Weigh Buick ginseng medicinal powder, add 10 times of volume 90% ethanol, heating and refluxing extraction 4 times, extraction time 3h, receive
Collection medicinal residues are placed in ventilation and dry, and dried medicinal residues are placed in round-bottomed flask, add 24 times of volume of water 90 DEG C and extract 4 times, carry
The time of taking is 3h, merges water and carries filtrate, and drying under reduced pressure, to thick, add the dehydrated alcohol of 5 times of volumes, puts in 4 DEG C of refrigerators
Stand 24h, have white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, 50 DEG C of drying under reduced pressure to weight,
Obtain.After measured, Buick gracilis polysaccharide yield: 11.45%.
Embodiment 5
The extraction preparation method of the Buick gracilis polysaccharide of the present invention is as follows:
Weigh Buick ginseng medicinal powder, add 12 times of volume 95% ethanol, heating and refluxing extraction 5 times, extraction time 4h, receive
Collection medicinal residues are placed in ventilation and dry, and dried medicinal residues are placed in round-bottomed flask, add 30 times of volume of water 100 DEG C and extract 5 times, carry
The time of taking is 4h, merges water and carries filtrate, and drying under reduced pressure, to thick, add the dehydrated alcohol of 6 times of volumes, puts in 4 DEG C of refrigerators
Stand 24h, have white polysaccharide to separate out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, 50 DEG C of drying under reduced pressure to weight,
Obtain.After measured, Buick gracilis polysaccharide yield: 12.23%.
Embodiment 6
The anti-inflammatory and antalgic test of Buick gracilis polysaccharide is as follows:
1, mice caused by dimethylbenzene xylene auricle edema test
Taking 50 mices and be randomly divided into 5 groups, often group 10, male, i.e. normal saline group, amoxicillin positive controls
(0.20 g/kg), Buick gracilis polysaccharide basic, normal, high dosage group, dosage is respectively 0.6,1.2,2.4g/kg, adaptability raises 3d.
Each group mice continuous gastric infusion 7d, dosage is 0.1mL/10g, 1 time/d, after last is administered 30min, positive and negative with mouse right ear
Face uniform application dimethylbenzene 50 μ L causes inflammation, and left ear compares, and after 30min puts to death, the dislocation of mice strength vertebra with the punching of diameter 8mm
Device lays round auricle, accurate weighing along the same position of left and right auricle.Result sees table 1.
Swelling rate (%)=((swelling ear weight-comparison ear weight)/comparison ear weight) × 100%；
Inhibitory rate of intumesce (%)=((the average swelling rate of normal saline group-average swelling rate of administration group)/normal saline group is average
Swelling rate) × 100%.
The impact of table 1 Buick gracilis polysaccharide xylol induced mice ear swelling
Note: compared with normal saline group,*P ＜ 0.01,*P ＜ 0.05；Compared with the group of amoxicillin,ΔΔP ＜ 0.01,ΔP
＜ 0.05
Data are through homogeneity test of variance, statistic of test F=2.473, P=0.058.Variance is neat, uses LSD method to carry out group
Between compare two-by-two.Result is as shown in table 1, and experimental data shows, compared with normal saline group, Buick gracilis polysaccharide can suppress dimethylbenzene
The mice ear caused, the biggest suppression ratio of dosage is the strongest, and suppression ratio presents good dose-effect relationship, wherein Buick with dosage
Gracilis polysaccharide high dose group suppression ratio reaches 44.47%, with normal saline group than significant difference (P=0.001)；Right with the aspirin positive
Comparing according to group, Buick gracilis polysaccharide high dose group swelling substantially reduces, significant difference (P=0.005).Prompting Buick gracilis polysaccharide tool
There is the acutely inflamed effect of certain intervention.
2, mice granuloma induced by implantation of cotton pellets test
Taking 50 mices and be randomly divided into 5 groups, often group 10, male, and packet and often group dosage are ibid.Adaptability raises 3d.
At ether light anaesthesia hypogastric region unhairing, abdominal incision, sterilizing cotton balls (about 25mg) is implanted mouse armpit subcutaneous.The same day, gavage was given
Medicine, continuous 7 d, dosage is 0.1mL/10g, 1 time/d, 1h after last administration, and cervical dislocation is put to death, and takes out cotton balls, does for 60 DEG C
Dry 24h, with scales/electronic balance weighing, deducts raw cotton ball and the most i.e. obtains cotton balls granulation tissue quality.Result sees table 2.
Granulation tissue quality=cotton balls granulation dry weight-cotton balls quality；
Suppression ratio=((normal saline group average granulation tissue quality-administration group average granulation tissue quality)/normal saline
Organize average granulation tissue quality) × 100%.
The impact on mice granuloma induced by implantation of cotton pellets of the table 2 Buick gracilis polysaccharide
Note: compared with normal saline group,*P ＜ 0.01,*P ＜ 0.05；Compared with the group of amoxicillin,ΔΔP ＜ 0.01,ΔP
＜ 0.05
Data are through homogeneity test of variance, statistic of test F=0.568, P=0.687, and variance is neat.LSD method is used to carry out group
Between compare two-by-two, result is as shown in table 2, and experimental data shows, compared with normal saline group, Buick gracilis polysaccharide each dosage equal energy of group
Substantially suppress the quality (P=0.008,0.000,0.000) of granuloma induced by implantation of cotton pellets, and the biggest suppression ratio of dosage is the strongest.Prompting Buick ginseng
Polysaccharide has certain effect intervening subacute inflammation.
3, glacial acetic acid causes mouse writhing test
Taking 50 mices and be randomly divided into 5 groups, often group 10, male, i.e. normal saline group, aspirin positive controls
(0.13 g/kg), Buick gracilis polysaccharide basic, normal, high dosage group, dosage is respectively 0.6,1.2,2.4g/kg, adaptability raises 3d.
Each group mice continuous gastric infusion 7d, dosage is 0.1mL/10g, is administered about 1h with last, each mice lumbar injection respectively
0.8% glacial acetic acid 0.1mL/10g, observes and records the writhing number of times that mice in 15min causes because of pain.Result sees table 3.
Suppression ratio=((normal saline group group average writhing number of times-administration group average writhing number of times)/normal saline group is average
Writhing number of times) × 100%.
Table 3 Buick gracilis polysaccharide causes the impact of mouse writhing to glacial acetic acid
Note: compared with normal saline group,*P ＜ 0.01,*P ＜ 0.05；Compared with aspirin group,ΔΔP ＜ 0.01,ΔP
＜ 0.05
Data are through homogeneity test of variance, statistic of test F=1.318, P=0.278, and variance is neat, use LSD method to carry out group
Between compare two-by-two.Result is as shown in table 3, and experimental data shows, compared with normal saline group, and Buick gracilis polysaccharide middle and high dosage group
Can obviously reduce the writhing number of times (P=0.008, P=0.001) of mice.Prompting Buick gracilis polysaccharide has certain analgesic activity.
In embodiment 6, the data processing method of each group test is as follows:
Application SPSS16.0 statistical software carries out Data Management Analysis, and result represents with mean ± standard deviation.Neat through variance
Property inspection after carry out one factor analysis of variance (one-way ANOVA), significance level takes α=0.05, poor between the group with P < 0.05
Different being judged as has statistical significance.
Finally it is noted that the foregoing is only the preferred embodiments of the present invention, it is not limited to the present invention,
Although being described in detail the present invention with reference to previous embodiment, for a person skilled in the art, it still may be used
So that the technical scheme described in foregoing embodiments to be modified, or wherein portion of techniques feature is carried out equivalent.
All within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included in the present invention's
Within protection domain.
Claims (6)
1. Buick gracilis polysaccharide application in preparation antiinflammatory, analgesic.
Application the most according to claim 1, it is characterised in that: the preparation method of described Buick gracilis polysaccharide is: be by Buick
After ginseng medicinal powder alcohol extraction defat, water extract-alcohol precipitation obtains Buick gracilis polysaccharide.
Application the most according to claim 2, it is characterised in that: described alcohol extraction is for using ethanol extraction.
4. according to the arbitrary described application of claim 1-3, it is characterised in that: the preparation method of described Buick gracilis polysaccharide is: claim
Take Buick ginseng medicinal powder, add 6-12 times of volume 70-100% ethanol, heating and refluxing extraction 1-5 time, extraction time 1-4h, receive
Collection medicinal residues dry, and add 6-30 times of volume of water 60-100 DEG C and extract 1-5 time in medicinal residues after the drying, and extraction time is 1-4h,
Merging water and carry filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 2-6 times of volume, and cold preservation stands, and has white polysaccharide to analyse
Going out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone cyclic washing, drying under reduced pressure, to weight, to obtain final product.
Application the most according to claim 4, it is characterised in that: the preparation method of described Buick gracilis polysaccharide is: weigh Buick
Ginseng medicinal powder, adds 12 times of volume 95% ethanol, heating and refluxing extraction 5 times, extraction time 4h, collects medicinal residues and dries, and is being dried
After medicinal residues in add 30 times of volume of water 100 DEG C and extract 5 times, extraction time is 4h, merges water and carries filtrate, and drying under reduced pressure is to thickness
Shape, adds the dehydrated alcohol of 6 times of volumes, and cold preservation stands, and has white polysaccharide to separate out, and filtration under diminished pressure, alcohol hypostasis ethanol, acetone are anti-
After backwashing is washed, and 50 DEG C of drying under reduced pressure, to weight, to obtain final product.
6. antiinflammatory, an analgesic, its effective ingredient is Buick gracilis polysaccharide.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20140041187A (en) * | 2012-09-27 | 2014-04-04 | 재단법인 전남생물산업진흥원 | Anti-cancer composition containing erythronium japonicum extract |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPS5529927A (en) * | 1978-08-22 | 1980-03-03 | Nakano Vinegar Co Ltd | Production of soy or seasoner including the same |
| JP2007308438A (en) * | 2006-05-19 | 2007-11-29 | Doctor's Bio Laboratory Co Ltd | Method for extracting essence and cosmetic |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140041187A (en) * | 2012-09-27 | 2014-04-04 | 재단법인 전남생물산업진흥원 | Anti-cancer composition containing erythronium japonicum extract |
Non-Patent Citations (1)
| Title |
|---|
| 新疆哈萨克民间药材别克参化学成分预实验;陈春丽等;《时珍国医国药》;20140520;第25卷(第5期);第1095-1097页 * |
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