CN104130338A - Preparation method of erythronium sibiricum polysaccharide and application of erythronium sibiricum - Google Patents

Preparation method of erythronium sibiricum polysaccharide and application of erythronium sibiricum Download PDF

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Publication number
CN104130338A
CN104130338A CN201410382303.4A CN201410382303A CN104130338A CN 104130338 A CN104130338 A CN 104130338A CN 201410382303 A CN201410382303 A CN 201410382303A CN 104130338 A CN104130338 A CN 104130338A
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buick
polysaccharide
preparation
alcohol
extraction
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CN104130338B (en
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热娜·卡斯木
陈春丽
李新霞
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Xinjiang Medical University
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Xinjiang Medical University
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Abstract

The invention provides a preparation method of an erythronium sibiricum polysaccharide. Erythronium sibiricum medicinal powder is subjected to water extraction and alcohol precipitation after being subjected to alcohol extraction and degreasing, so as to obtain the erythronium sibiricum polysaccharide. The invention also provides an application of the erythronium sibiricum polysaccharide in preparation of anti-inflammatory and analgesic drugs. The erythronium sibiricum polysaccharide has good effects of diminishing inflammation and easing pain by an experiment, and can be applied to preparation of the anti-inflammatory and analgesic drugs.

Description

The preparation method of Buick gracilis polysaccharide and application thereof
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to preparation method and the application thereof of Buick gracilis polysaccharide.
Background technology
Buick ginseng is the Liliaceae pig tartar platymiscium Xinjiang pig tartar dry meat bulb of (having another name called siberian pig tartar, Altai Mountains pig tartar).Originate from In The North of Xinjiang (Tianshan Area is to Altay, Fu Hai), be born under Schattenseite, leaf margin thick grass, sylvan life, thicket and on sub-alpine meadow height above sea level 1100-2500m.Also there is distribution in siberian to Central Asia.Excavate June in the time that cauline leaf is not yet withered, and boiling water dries after slightly boiling.Property dry, micro-hardship, nourish, keep fit, strong body, how sickly cure mainly body void, soreness of the waist and knees, unable, is one of time-honored medicine food dual purpose plant of Xinjiang Kazak nationality.
Inflammation refer to have that damage that the biological tissue of vascular system causes various pro-inflammatory cytokines occurs taking defensive raction as main pathologic process.That basic pathology is changed to is rotten, ooze out and hyperplasia.Topical manifestations is red, swollen, hot, pain and dysfunction, and systemic reaction has in heating, blood White blood cell etc.Inflammation is found in the pathologic process of various diseases.Anti-inflammatory drug refers to a class medicine that inflammatory reaction is had to restraining effect or conciliation effect.The anti-inflammatory action of drugs, seeking safe and effective anti-inflammatory substance is the common problems of paying close attention to of numerous medical scholars.The anti-inflammatory action of drugs, must first copy inflammatory animal model, chooses multiple model and carries out anti-inflammatory screening, to reach the object of drug effect being carried out to comprehensive evaluation.
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, causes some inflammatory mediator as histamine, kassinin kinin and fibrinolysis discharge, and causes local vessel expansion, and capillary permeability increases, and inflammatory cell infiltration, causes ear's acute exudative inflammation oedema.The difference of ear swelling degree after comparative experiments group and the effect of control group dimethylbenzene, to determine whether given the test agent has the effect of intervention acute inflammation (mice ear).
Mouse granuloma induced by implantation of cotton pellets experimental model is typical subacute inflammation model.Pathologic Characteristics infiltrates as main taking lymphocyte and mononuclear macrophage, has the hyperplasia of the parenchymas such as obvious fibrillar connective tissue, blood vessel and epithelial cell, i.e. granulation.Generate the difference of granulation tissue quality by comparative experiments group and control group, to determine whether given the test agent has the effect of intervention subacute inflammation (mouse granuloma induced by implantation of cotton pellets).
It is one of method of checking medicine analgesic effect that Glacial acetic acid causes mouse writhing experimental model.Dominant mechanism is that the chemical irritants of a constant volume and concentration is injected in mouse peritoneal, stimulate visceral layer and parietal peritoneum, cause the inflammatory pain of larger area, long period, cause mouse to occur belly indent, trunk and hind leg extension, the behavior reactions such as hips up, are called writhing response.This reaction after injection in 15min frequency higher, therefore to inject the writhing number of times occurring in rear 15min as pain quantitative target.By the difference of comparative experiments group and control 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 in the application of preparing in anti-inflammation analgesis medicament.
The invention provides a kind of preparation method of Buick gracilis polysaccharide, is that Buick is joined after medicinal powder alcohol extracting degreasing, and water extract-alcohol precipitation obtains Buick gracilis polysaccharide.
Preferably, described alcohol extracting is ethanol.
Preferably, described method is: 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, collects the dregs of a decoction and dries, and adds 60-100 DEG C, 6-30 times of volume water to extract in the dried dregs of a decoction 1-5 time, extraction time is 1-4h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 2-6 times of volume, refrigeration leaves standstill, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, drying under reduced pressure, to weight, to obtain final product.
Most preferably, described method is: take Buick ginseng medicinal powder, add 12 times of volume 95% ethanol, heating and refluxing extraction 5 times, extraction time 4h, collects the dregs of a decoction and dries, in the dried dregs of a decoction, add 100 DEG C, 30 times of volume water to extract 5 times, extraction time is 4h, merges water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 6 times of volumes, and refrigeration leaves standstill, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.
The present invention also provides the application Buick gracilis polysaccharide that above-mentioned method prepares.
The 3rd object of the present invention is to provide the application of Buick gracilis polysaccharide in preparation anti-inflammatory, analgesic.
The 4th object of the present invention is to provide a kind of anti-inflammatory, analgesic, and its effective constituent is Buick gracilis polysaccharide.
Through test, Buick gracilis polysaccharide has good effect to anti-inflammatory, analgesia, can be used in the medicine of preparation anti-inflammatory, analgesia class.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.
1 material
1.1 medicinal material
Buick ginseng medicinal material picks up from Altay, Xinjiang, is accredited as the bulb of liliaceous plant Erythronium sibiricum (Fisch. et. Mey.) Kryl Xinjiang pig tartar through pharmaceutical college of Xinjiang Medicine University.
1.2 animal
Kunming mice, body weight 18-22g, male, provided by Xinjiang Medicine University's experimental animal center, normally raises and start test after 3d.
1.3 reagent
Dehydrated alcohol, acetone, acetylsalicylic acid effervescent tablet (AstraZeneca pharmaceutical Co. Ltd, lot identification mark 1208042), amoxil capsule (Baiyunshan Pharmaceutical General Factory, lot identification mark 4190047), dimethylbenzene, Glacial acetic acid, ether.
1.4 equipment
Water-bath, Rotary Evaporators, vacuum drying oven, analytical balance, punch tool.
embodiment 1
The extraction preparation method of Buick gracilis polysaccharide of the present invention is as follows:
Take Buick ginseng medicinal powder, add 6 times of volume 70% ethanol, heating and refluxing extraction 1 time, extraction time 1h, the collection dregs of a decoction are placed in ventilation and dry, and the dried dregs of a decoction are placed in round-bottomed flask, add 60 DEG C, 6 times of volume water to extract 1 time, extraction time is 1h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 2 times of volumes, put into 4 DEG C of refrigerators and leave standstill 24h, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.After measured, Buick gracilis polysaccharide yield: 3.12%.
embodiment 2
The extraction preparation method of Buick gracilis polysaccharide of the present invention is as follows:
Take Buick ginseng medicinal powder, add 7 times of volume 80% ethanol, heating and refluxing extraction 2 times, extraction time 2h, the collection dregs of a decoction are placed in ventilation and dry, and the dried dregs of a decoction are placed in round-bottomed flask, add 70 DEG C, 12 times of volume water to extract 2 times, extraction time is 2h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 3 times of volumes, put into 4 DEG C of refrigerators and leave standstill 24h, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.After measured, Buick gracilis polysaccharide yield: 6.24%.
embodiment 3
The extraction preparation method of Buick gracilis polysaccharide of the present invention is as follows:
Take Buick ginseng medicinal powder, add 8 times of volume 80% ethanol, heating and refluxing extraction 3 times, extraction time 3h, the collection dregs of a decoction are placed in ventilation and dry, and the dried dregs of a decoction are placed in round-bottomed flask, add 80 DEG C, 18 times of volume water to extract 3 times, extraction time is 3h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 4 times of volumes, put into 4 DEG C of refrigerators and leave standstill 24h, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.After measured, Buick gracilis polysaccharide yield: 9.38%.
embodiment 4
The extraction preparation method of Buick gracilis polysaccharide of the present invention is as follows:
Take Buick ginseng medicinal powder, add 10 times of volume 90% ethanol, heating and refluxing extraction 4 times, extraction time 3h, the collection dregs of a decoction are placed in ventilation and dry, and the dried dregs of a decoction are placed in round-bottomed flask, add 90 DEG C, 24 times of volume water to extract 4 times, extraction time is 3h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 5 times of volumes, put into 4 DEG C of refrigerators and leave standstill 24h, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.After measured, Buick gracilis polysaccharide yield: 11.45%.
embodiment 5
The extraction preparation method of Buick gracilis polysaccharide of the present invention is as follows:
Take Buick ginseng medicinal powder, add 12 times of volume 95% ethanol, heating and refluxing extraction 5 times, extraction time 4h, the collection dregs of a decoction are placed in ventilation and dry, and the dried dregs of a decoction are placed in round-bottomed flask, add 100 DEG C, 30 times of volume water to extract 5 times, extraction time is 4h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 6 times of volumes, put into 4 DEG C of refrigerators and leave standstill 24h, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.After measured, Buick gracilis polysaccharide yield: 12.23%.
embodiment 6
The relieving inflammation and relaxing pain test of Buick gracilis polysaccharide is as follows:
1, mice caused by dimethylbenzene xylene auricle edema test
Get 50 mouse and be divided at random 5 groups, 10 every group, male, i.e. physiological saline group, amoxycilline Trihydrate bp positive controls (0.20 g/kg), the basic, normal, high dosage group of Buick gracilis polysaccharide, dosage is respectively 0.6,1.2,2.4g/kg, and adaptability is raised 3d.The continuous gastric infusion 7d of each group mouse, dosage is 0.1mL/10g, 1 time/d, after last administration 30min, evenly smear dimethylbenzene 50 μ L with mouse right ear pros and cons and cause inflammation, left ear contrasts, and after 30min, the dislocation of mouse strength vertebra is put to death, lay round auricle, accurate weighing along the same position of left and right auricle with the punch tool of diameter 8mm.Result is referring to table 1.
Swelling rate (%)=((swelling ear weight-contrast ear weight)/contrast ear weight) × 100%;
Inhibitory rate of intumesce (%)=((the average swelling rate of the average swelling rate-administration of physiological saline group group)/average swelling rate of physiological saline group) × 100%.
The impact of table 1 Buick gracilis polysaccharide p-Xylol induced mice ear swelling
Note: compared with physiological saline group, *p < 0.01, *p < 0.05; Compared with the group of amoxycilline Trihydrate bp, Δ Δp < 0.01, Δp < 0.05
Data are through homogeneity test of variance, test statistic F=2.473, P=0.058.Variance is neat, and employing LSD method compares between organizing between two.Result is as shown in table 1, experimental data shows, compared with physiological saline group, Buick gracilis polysaccharide can suppress the mice ear that dimethylbenzene causes, the larger inhibiting rate of dosage is stronger, and inhibiting rate and dosage present good dose-effect relationship, wherein Buick gracilis polysaccharide high dose group inhibiting rate reaches 44.47%, with physiological saline group than significant difference (P=0.001); Compared with acetylsalicylic acid positive controls, Buick gracilis polysaccharide high dose group swelling obviously reduces, significant difference (P=0.005).Prompting Buick gracilis polysaccharide has the acutely inflamed effect of certain intervention.
2, mouse granuloma induced by implantation of cotton pellets test
Get 50 mouse and be divided at random 5 groups, 10 every group, male, grouping and every group of dosage are the same.Adaptability is raised 3d.At ether light anaesthesia abdomen unhairing, abdominal incision, implants mouse armpit by sterilizing cotton balls (about 25mg) subcutaneous.The same day gastric infusion, continuous 7 d, dosage is 0.1mL/10g, 1 time/d, 1h after last administration, cervical vertebra dislocation is put to death, and takes out cotton balls, 60 DEG C of dry 24h, with scales/electronic balance weighing, deduct former cotton balls and heavily obtain cotton balls granulation tissue quality.Result is referring to table 2.
Granulation tissue quality=cotton balls granulation dry weight quality-cotton balls quality;
Inhibiting rate=((the average granulation tissue quality of the average granulation tissue quality-administration of physiological saline group group) the average granulation tissue quality of/physiological saline group) × 100%.
The impact of table 2 Buick gracilis polysaccharide on mouse granuloma induced by implantation of cotton pellets
Note: compared with physiological saline group, *p < 0.01, *p < 0.05; Compared with the group of amoxycilline Trihydrate bp, Δ Δp < 0.01, Δp < 0.05
Data are through homogeneity test of variance, test statistic F=0.568, and P=0.687, variance is neat.Employing LSD method compares between organizing between two, and result is as shown in table 2, and experimental data shows, compared with physiological saline group, the each dosage group of Buick gracilis polysaccharide all can obviously suppress quality (P=0.008,0.000 of granuloma induced by implantation of cotton pellets, 0.000), and the larger inhibiting rate of dosage stronger.Prompting Buick gracilis polysaccharide has the effect of certain intervention subacute inflammation.
3, Glacial acetic acid causes mouse writhing test
Get 50 mouse and be divided at random 5 groups, 10 every group, male, i.e. physiological saline group, acetylsalicylic acid positive controls (0.13 g/kg), the basic, normal, high dosage group of Buick gracilis polysaccharide, dosage is respectively 0.6,1.2,2.4g/kg, and adaptability is raised 3d.The continuous gastric infusion 7d of each group mouse, dosage is 0.1mL/10g, with the about 1h of last administration, each mouse is abdominal injection 0.8% Glacial acetic acid 0.1mL/10g respectively, observes and record the writhing number of times that in 15min, mouse causes because of pain.Result is referring to table 3.
Inhibiting rate=((the average writhing number of times of the average writhing number of times-administration of physiological saline group group group) the average writhing number of times of/physiological saline group) × 100%.
Table 3 Buick gracilis polysaccharide causes the impact of mouse writhing on Glacial acetic acid
Note: compared with physiological saline group, *p < 0.01, *p < 0.05; Compared with acetylsalicylic acid group, Δ Δp < 0.01, Δp < 0.05
Data are through homogeneity test of variance, test statistic F=1.318, and P=0.278, variance is neat, adopts LSD method to compare between two between organizing.Result is as shown in table 3, experimental data demonstration, and compared with physiological saline group, the middle and high dosage group of Buick gracilis polysaccharide can obviously reduce the writhing number of times (P=0.008, P=0.001) of mouse.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.After homogeneity test of variance, carry out one-way analysis of variance (one-way ANOVA), significance level is got α=0.05, is judged as and has statistical significance with the group difference of P<0.05.
Finally it should be noted that: the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although the present invention is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement.Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.

Claims (7)

1. a preparation method for Buick gracilis polysaccharide, is characterized in that: be that Buick is joined after medicinal powder alcohol extracting degreasing, water extract-alcohol precipitation obtains Buick gracilis polysaccharide.
2. preparation method according to claim 1, is characterized in that: described alcohol extracting is ethanol.
3. preparation method according to claim 1, it is characterized in that: described method is: 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, the collection dregs of a decoction dry, in the dried dregs of a decoction, add 60-100 DEG C, 6-30 times of volume water to extract 1-5 time, extraction time is 1-4h, merge water extraction filtrate, drying under reduced pressure is to thick, add the dehydrated alcohol of 2-6 times of volume, refrigeration leaves standstill, adularescent polysaccharide is separated out, filtration under diminished pressure, alcohol hypostasis ethanol, acetone repetitive scrubbing, drying under reduced pressure is to weight, obtain.
4. preparation method according to claim 3, it is characterized in that: described method is: take Buick ginseng medicinal powder, add 12 times of volume 95% ethanol, heating and refluxing extraction 5 times, extraction time 4h, collects the dregs of a decoction and dries, and adds 100 DEG C, 30 times of volume water to extract in the dried dregs of a decoction 5 times, extraction time is 4h, merge water extraction filtrate, drying under reduced pressure, to thick, adds the dehydrated alcohol of 6 times of volumes, refrigeration leaves standstill, adularescent polysaccharide is separated out, filtration under diminished pressure, and alcohol is ethanol, acetone repetitive scrubbing for hypostasis, 50 DEG C of drying under reduced pressure, to weight, to obtain final product.
5. application rights requires the Buick gracilis polysaccharide that the arbitrary described method of 1-4 prepares.
6. the application of Buick gracilis polysaccharide in preparation anti-inflammatory, analgesic.
7. anti-inflammatory, an analgesic, its effective constituent is Buick gracilis polysaccharide.
CN201410382303.4A 2014-08-06 2014-08-06 The preparation method and applications of Buick gracilis polysaccharide Expired - Fee Related CN104130338B (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
KR20140041187A (en) * 2012-09-27 2014-04-04 재단법인 전남생물산업진흥원 Anti-cancer composition containing erythronium japonicum extract

Non-Patent Citations (3)

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王如峰等: "鄂产竹节参多糖的抗炎、镇痛活性", 《中国医院药学杂志》 *
陈春丽等: "新疆哈萨克民间药材别克参化学成分预实验", 《时珍国医国药》 *
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