CN1273493C - Angelica polysaccharide and its preparation and use - Google Patents
Angelica polysaccharide and its preparation and use Download PDFInfo
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- CN1273493C CN1273493C CN 200410061139 CN200410061139A CN1273493C CN 1273493 C CN1273493 C CN 1273493C CN 200410061139 CN200410061139 CN 200410061139 CN 200410061139 A CN200410061139 A CN 200410061139A CN 1273493 C CN1273493 C CN 1273493C
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Abstract
The present invention relates to angelica polysaccharide which is prepared with a method which comprises the following steps of dissolving angelica alcohol precipitates into distilled water, removing protein with a Sevag method until protein adsorption peaks of 280 nm do not exist through ultraviolet detection, centrifuging and removing precipitates, reserving water phase, adding anhydrous alcohol, precipitating polysaccharide, discarding supernatant fluid, washing the obtained precipitates by anhydrous alcohol, freezing and drying the precipitates, separating gelatin molecules in an exclusion chromatography mode, collecting separated products with relative molecular mass range of 10000 to 110000, and freezing and drying the separated products to obtain the angelica polysaccharide. The angelica polysaccharide of the present invention has obvious analgesic effect on pain caused by chemical substances and thermal stimulation, and can effectively resist pain reaction caused by dysmenorrhoea induced by female hormone. The angelica polysaccharide can be used for preparing analgesic medicines, especially medicines for treating dysmenorrhea.
Description
Technical field
The present invention relates to a kind of Radix Angelicae Sinensis polysaccharide and its production and use.
Background technology
The angelica tonic is the dry root of umbelliferae (umbuiferae) angelica per nnial herb Radix Angelicae Sinensis [AngelicaSinensis (Oliv) Diels].Radix Angelicae Sinensis has benefiting blood and regulating blood circulation, and is promoting blood circulation and removing blood stasis, and the effect of menstruction regulating and pain relieving is often used Radix Angelicae Sinensis and other medicines compatibility according to disease clinically, is used for diseases such as menoxenia, dysmenorrhoea, headache due to deficiency of blood, waist abdomen pain.
The chemical constitution of Radix Angelicae Sinensis comprises volatile oil and water soluble ingredient two parts.Early stage research was once pointed out Radix Angelicae Sinensis oil to various animal isolated uterines, no matter the person of becoming pregnant or unpregnancy person have the effect of relaxing.It has made explanation for Radix Angelicae Sinensis is used for dysmenorrhea treatment to a certain extent.
Radix Angelicae Sinensis polysaccharide is present in the Radix Angelicae Sinensis water soluble part, the homology mixture that it is made up of the different macromolecular compound of molecular weight.In recent years, Radix Angelicae Sinensis polysaccharide has been proved to be has complicated biological activity, can regulate body's immunological function, the caused inhibition of antagonism cortin, prevent the minimizing of peripheral leukocytes and the irradiation ultimate effect in the radiation injury, enhancing body hemopoietic function and antitumor action.
Pain is common clinical symptom, and seek non-safely and effectively addicted analgesics thing is the problem that the medicine worker attaches great importance to always." dysmenorrhoea " be common gynecological disease especially, and the traditional Chinese medical science thinks that its pathogeny mainly is a QI and blood being obstructed, due to the hematogenous blockage.How Radix Angelicae Sinensis searches out its effective therapeutic component to carry out modernization of Chinese medicine reform as gynaecology's key medicine of the traditional Chinese medical science, makes it meet the modern formulation requirement, is the research focus of medicine educational circles.
" Chinese Pharmaceutical Journal " 2002; 37 (10): 746-8 " research of Radix Angelicae Sinensis Crude polysaccharides experiment of analgesic " discloses the Radix Angelicae Sinensis Crude polysaccharides and has analgesic activity, but sugared content is relatively low in the used Radix Angelicae Sinensis Crude polysaccharides, also contain other composition, and analgesic effect is not obvious.
Summary of the invention
Problem to be solved by this invention provides a kind of Radix Angelicae Sinensis polysaccharide preparation method and purposes, and the gained Radix Angelicae Sinensis polysaccharide has obvious analgesic activity, and especially treatment " dysmenorrhoea " has obvious effects.
Technical scheme provided by the invention is: a kind of Radix Angelicae Sinensis polysaccharide is made by laxative remedy: the angelol hypostasis is dissolved in distilled water, does not have the protein absorption peak of 280nm, the centrifugal precipitation of removing with Sevag method Deproteinization until ultraviolet detection; Keep water, add dehydrated alcohol, the precipitation polysaccharide is abandoned supernatant liquor, the gained throw out is with absolute ethanol washing, and after lyophilize, the gel molecular exclusion chromatography separates, and collects the relative molecular mass scope 1,0000-11, the separated product between 0000 obtains Radix Angelicae Sinensis polysaccharide again after lyophilize.
The present invention also provides the preparation method of above-mentioned Radix Angelicae Sinensis polysaccharide, and the angelol hypostasis is dissolved in distilled water, does not have the protein absorption peak of 280nm, the centrifugal precipitation of removing with Sevag method Deproteinization until ultraviolet detection; Keep water, add dehydrated alcohol, the precipitation polysaccharide is abandoned supernatant liquor, the gained throw out is colourless with absolute ethanol washing to washings, discard washings, the throw out after the washing is through lyophilize, and the gel molecular exclusion chromatography separates, collect the relative molecular mass scope 1,0000-11, the separated product between 0000 obtains Radix Angelicae Sinensis polysaccharide again after lyophilize.
Above-mentioned raw materials angelol hypostasis among the present invention can be used extracting method gained angelol hypostasis always from Chinese medicinal materials in the Chinese medicine preparation process: promptly Radix Angelicae Sinensis obtains water extract through poach, the throw out of water extract behind extraction using alcohol.
The angelol hypostasis also can be obtained by laxative remedy: get angelica sinensis, the decocting that adds 5～7 times of angelica sinensis weight boiled 30～60 minutes, filtered; The water that the gained dregs of a decoction is added 3～6 times of weight (dregs of a decoction) decocted 30～50 minutes, filtered; The decocting that the gained dregs of a decoction add 3～6 times of weight (dregs of a decoction) boiled 30～50 minutes, filtered; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 40%～60% (volume ratio, down together), and 2～4 ℃ refrigerate 24～48 hours, filter, and get throw out; Concentrate once more, adding ethanol to ethanol content is 70%～80%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate for the third time, adding ethanol to ethanol content is 80%～90%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Merge three throw outs and get Radix Angelicae Sinensis alcohol precipitation thing.
The resulting above-mentioned Radix Angelicae Sinensis polysaccharide of the present invention belongs to polymkeric substance, the representative of its molecular weight be similar chain length on average join cloth.Physico-chemical property through identifying Radix Angelicae Sinensis polysaccharide of the present invention is: the relative molecular mass scope is 1,0000-11, (adopt gel permeation chromatography-laser light scattering coupling technique) between 0000, soluble in water, sugar content is greater than 95%, detect through HPLC and to show mainly and forms, and ultra-violet absorption spectrum demonstration 192nm place there is obvious absorption peaks by glucose, pectinose, semi-lactosi, rhamnosyl and wood sugar; Infrared absorption spectrum analysis is at 3352cm
-1, 2940cm
-1, 1747cm
-1, 1626cm
-1, 1414cm
-1, 1237cm
-1, 1021cm
-1And 536cm
-1The place shows as typical polysaccharide absorption peak.
Above-mentioned Radix Angelicae Sinensis polysaccharide of the present invention is especially treated purposes in the dysmenorrhoea medicine at the preparation analgesic.As Radix Angelicae Sinensis polysaccharide of the present invention is used to prepare capsule, tablet, granule and injection, the gained medicine has significant analgesia role, and especially treatment " dysmenorrhoea " effect is remarkable.
During by the disposable oral administration 0.25g/kg of the Radix Angelicae Sinensis Crude polysaccharides in the background technology (sugared content is 65.5%), chemical substance and thermal stimulus are caused that the model of pain has obvious analgesic activity, and Radix Angelicae Sinensis polysaccharide of the present invention can produce suitable effect when disposable oral administration 0.025g/kg, using dosage descends 90%, and increases the analgesic activity enhancing with dosage.
Embodiment
Below in conjunction with concrete example technical scheme of the present invention is described further:
Above-mentioned raw materials angelol hypostasis among the present invention can be used extracting method gained angelol hypostasis always from Chinese medicinal materials in the Chinese medicine preparation process: promptly Radix Angelicae Sinensis obtains water extract through poach, the throw out of water extract behind extraction using alcohol.
The angelol hypostasis also can be obtained by laxative remedy: get angelica sinensis, the decocting that adds 5～7 times of angelica sinensis weight boiled 30～60 minutes, filtered; The water that the gained dregs of a decoction is added 3～6 times of weight (dregs of a decoction) decocted 30～50 minutes, filtered; The decocting that the gained dregs of a decoction add 3～6 times of weight (dregs of a decoction) boiled 30～50 minutes, filtered; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 40%～60% (volume ratio, down together), and 2～4 ℃ refrigerate 24～48 hours, filter, and get throw out; Concentrate once more, adding ethanol to ethanol content is 70%～80%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate for the third time, adding ethanol to ethanol content is 80%～90%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Merge three throw outs and get Radix Angelicae Sinensis alcohol precipitation thing.
The preparation of Radix Angelicae Sinensis polysaccharide of the present invention: the angelol hypostasis is dissolved in distilled water, and adding ethanol to ethanol content is 70%～80% extracting, and 2～4 ℃ refrigerate 24～48 hours, abandon supernatant, keep precipitation, and it is colourless until supernatant liquor to repeat this step, keeps precipitation.To precipitate once more and be dissolved in distilled water again, not have the protein absorption peak of 280nm, the centrifugal metaprotein of removing until ultraviolet detection with Sevag method Deproteinization.Keep water, add the dehydrated alcohol of 3～4 times of volumes,, abandon supernatant in 2～4 ℃ of refrigerations 24～48 hours.Precipitation part is colourless with absolute ethanol washing to washings, discards washings, treat that ethanol volatilizees naturally after, lyophilize 5～20 hours, products therefrom belongs to polymkeric substance, its molecular weight is represented be similar chain length on average join cloth.Separate through the gel molecular exclusion chromatography, collect the relative molecular mass scope 1,0000-11, the separated product between 0000 obtains Radix Angelicae Sinensis polysaccharide again after lyophilize.Physico-chemical property through identifying gained Radix Angelicae Sinensis polysaccharide of the present invention is: the relative molecular mass scope is 1,0000-11, (adopt gel permeation chromatography-laser light scattering coupling technique) between 0000, soluble in water, sugar content is greater than 95%, detect through HPLC and to show mainly and forms, and ultra-violet absorption spectrum demonstration 192nm place there is obvious absorption peaks by glucose, pectinose, semi-lactosi, rhamnosyl and wood sugar; Infrared absorption spectrum analysis is at 3352cm
-1, 2940cm
-1, 1747cm
-1, 1626cm
-1, 1414cm
-1, 1237cm
-1, 1021cm
-1And 536cm
-1The place shows as typical polysaccharide absorption peak.
Embodiment 1: get 5 times of (weight ratio) decoctings of angelica sinensis adding and boiled 30 minutes, filter; The gained dregs of a decoction are added 3 times of (weight ratio) water decocted 30 minutes, filter; The gained dregs of a decoction add 3 times of (weight ratio) decoctings and boiled 30 minutes, filter; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 40% (v/v), and 4 ℃ refrigerate 24 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate once more, adding ethanol to ethanol content is 70% (v/v), and 4 ℃ refrigerate 24 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate for the third time, adding ethanol to ethanol content is 80% (v/v), and 4 ℃ refrigerate 24 hours, filter, and keeping precipitation is the angelol hypostasis.Merge precipitation and promptly merge the angelol hypostasis, it is dissolved in distilled water again, adding ethanol to ethanol content is 70% (v/v) extracting, and 4 ℃ refrigerate 24 hours, abandon supernatant, keep precipitation, and it is colourless until supernatant liquor to repeat this step, keeps precipitation.To precipitate once more and be dissolved in distilled water again, not have the protein absorption peak of 280nm, the centrifugal metaprotein of removing until ultraviolet detection with Sevag method Deproteinization.Keep water, add the dehydrated alcohol of 4 times of volumes,, abandon supernatant in 4 ℃ of refrigerations 24 hours.Precipitation part is colourless with absolute ethanol washing to washings, discards washings, treat that ethanol volatilizees naturally after, lyophilize 5 hours; With TSK-Gel 4000H type post (pore aperture 600 dusts), column dimension 250 * 10mm is a moving phase with distilled water, flow 10ml/min, the separated product of collection 5-60min; After lyophilize, obtain Radix Angelicae Sinensis polysaccharide again.Adopt gel permeation chromatography-laser light scattering coupling technique, HPLC and infrared absorption spectrum analysis identify that the yield of above-mentioned Radix Angelicae Sinensis polysaccharide is 10%, and purity is 96%.
Embodiment 2: get 7 times of (weight ratio) decoctings of angelica sinensis adding and boiled 60 minutes, filter; The gained dregs of a decoction are added 5 times of (weight ratio) water decocted 40 minutes, filter; The gained dregs of a decoction add 3 times of (weight ratio) decoctings and boiled 40 minutes, filter; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 60% (v/v), and 2 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate once more, adding ethanol to ethanol content is 80% (v/v), and 2 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate for the third time, adding ethanol to ethanol content is 80% (v/v), and 2 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis.Merge precipitation and promptly merge the angelol hypostasis, it is dissolved in distilled water again, adding ethanol to ethanol content is 80% (v/v) extracting, and 2 ℃ refrigerate 48 hours, abandon supernatant, keep precipitation, and it is colourless until supernatant liquor to repeat this step, keeps precipitation.To precipitate once more and be dissolved in distilled water again, not have the protein absorption peak of 280nm, the centrifugal metaprotein of removing until ultraviolet detection with Sevag method Deproteinization.Keep water, add the dehydrated alcohol of 3 times of volumes,, abandon supernatant in 2 ℃ of refrigerations 48 hours.Precipitation part is colourless with absolute ethanol washing to washings, discards washings, treat that ethanol volatilizees naturally after, lyophilize 10 hours; With TSK-Gel 4000H type post (pore aperture 600 dusts), column dimension 250 * 10mm is a moving phase with distilled water, flow 10ml/min, the separated product of collection 5-60min; After lyophilize, obtain Radix Angelicae Sinensis polysaccharide again.Adopt gel permeation chromatography-laser light scattering coupling technique, HPLC and infrared absorption spectrum analysis identify that the yield of above-mentioned Radix Angelicae Sinensis polysaccharide is 30%, and purity is 97%.
Embodiment 3: get 6 times of (weight ratio) decoctings of angelica sinensis adding and boiled 50 minutes, filter; The gained dregs of a decoction are added 6 times of (weight ratio) water decocted 50 minutes, filter: the gained dregs of a decoction add 6 times of (weight ratio) decoctings and boiled 50 minutes, filter; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 50% (v/v), and 4 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate once more, adding ethanol to ethanol content is 80% (v/v), and 4 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis; Concentrate for the third time, adding ethanol to ethanol content is 90% (v/v), and 4 ℃ refrigerate 48 hours, filter, and keeping precipitation is the angelol hypostasis.Merge precipitation and promptly merge the angelol hypostasis, it is dissolved in distilled water again, adding ethanol to ethanol content is 70% (v/v) extracting, and 4 ℃ refrigerate 48 hours, abandon supernatant, keep precipitation, and it is colourless until supernatant liquor to repeat this step, keeps precipitation.To precipitate once more and be dissolved in distilled water again, not have the protein absorption peak of 280nm, the centrifugal metaprotein of removing until ultraviolet detection with Sevag method Deproteinization.Keep water, add the dehydrated alcohol of 4 times of volumes,, abandon supernatant in 4 ℃ of refrigerations 24 hours.Precipitation part is colourless with absolute ethanol washing to washings, discards washings, treat that ethanol volatilizees naturally after, lyophilize 20 hours; With TSK-Gel 4000H type post (pore aperture 600 dusts), column dimension 250 * 10mm is a moving phase with distilled water, flow 10ml/min, the separated product of collection 5-60min: obtain Radix Angelicae Sinensis polysaccharide again after lyophilize.Adopt gel permeation chromatography-laser light scattering coupling technique, HPLC and infrared absorption spectrum analysis identify that the yield of above-mentioned Radix Angelicae Sinensis polysaccharide is 40%, and purity is 96%.
Embodiment 4: use each analgesic medicine of Radix Angelicae Sinensis polysaccharide system of the present invention: Radix Angelicae Sinensis polysaccharide adds vehicle, makes tablet according to a conventional method.
Embodiment 5: prepare the analgesic medicine with Radix Angelicae Sinensis polysaccharide of the present invention: Radix Angelicae Sinensis polysaccharide adds vehicle, makes capsule according to a conventional method.
Embodiment 6: prepare the analgesic medicine with Radix Angelicae Sinensis polysaccharide of the present invention: Radix Angelicae Sinensis polysaccharide is dissolved in the water for injection, makes injection according to a conventional method.
Animal experiment:
1. medicine: diethylstilbestrol injection, Shanghai the 9th pharmaceutical factory, lot number: 020906.Oxytocin inj, biochemical-pharmaceutical factory, Nanjing, lot number: 030409.
(2) key instrument GL-8402 type hot plate pain threshold detector, medicine instrument plant in Ninghai, Zhejiang produces.
(3) animal health Kunming mouse removes " dysmenorrhea model " and selects for use
Mouse is as outside the negative control, and the ♀ mouse is all used in other experiment, and body weight 18 ± 1g is provided by Wuhan University experimentation on animals center, conformity certification number: SCXK (Hubei Province) 2003-2004.
2. method
(1) influence of female mice writhing response due to the Radix Angelicae Sinensis polysaccharide Dichlorodiphenyl Acetate of the present invention
Animal grouping and drug dose see Table 1, test preceding 1 hour Radix Angelicae Sinensis Crude polysaccharides (sugared content is 65.5%) or the Radix Angelicae Sinensis polysaccharide of the present invention in the oral administration background technology, and model group waits capacity physiological saline.1 hour abdominal injection 1% acetic acid 0.2ml/ only observes in administration mouse writhing response latency and the 30min and turns round the body number of times after the administration, and calculates the protection percentage.
(2) Radix Angelicae Sinensis polysaccharide of the present invention is to the influence of hot plate method analgesic activity
With 55 ℃ of hot plates the ♀ mouse is screened in advance, the qualified mouse that is of foot reaction in 10～30 seconds, occurred licking.Experiment grouping and drug dose see Table 2.Testing preceding 1 hour per os irritates stomach (ig) and is subjected to reagent, model group to wait capacity physiological saline.For avoiding the influence of threshold of pain cyclic fluctuation, experiment all o'clock is carried out 25 ± 1 ℃ of room temperatures in the morning 8～10.The variation of the threshold of pain respectively organized in record, and calculate the threshold of pain and improve percentage.If sufficient reaction appears licking in the end in 60 seconds, the threshold of pain is counted 60 seconds.
(3) Radix Angelicae Sinensis polysaccharide of the present invention brings out the influence of Mouse Uterus spasm to stilboestrol and oxytocin
Each treated animal is subcutaneous injection stilboestrol 0.2mg/ before experiment, for three days on end.Test the same day, the animal per os is irritated stomach (ig) and is given Radix Angelicae Sinensis Crude polysaccharides or a kind of Radix Angelicae Sinensis polysaccharide of the present invention, and model group waits capacity physiological saline, and ip oxytocin 0.3u/ only after 1 hour.Observe in each treated animal writhing response latent period and the 30min and turn round the body number of times.If writhing response do not occur in the 30min, count 30min latent period.
3. result
(1) Radix Angelicae Sinensis polysaccharide of the present invention is to pain model---and the influence experiment of female mice writhing response due to the acetic acid the results are shown in Table 1.
Writhing response influences .n (routine number)=10, x ± s (mean ± standard deviation) due to the table 1 Radix Angelicae Sinensis polysaccharide Dichlorodiphenyl Acetate of the present invention
| Grouping | Dosage (gkg -1) | Writhing response | ||
| Latent period (min) | Number of times (inferior/30min) | Protection ratio (%) | ||
| A kind of Radix Angelicae Sinensis polysaccharide of model group Radix Angelicae Sinensis Crude polysaccharides | / 0.075 0.250 0.025 0.075 | 5.2±1.8 6.9±1.5 1) 19.3±6.8 2) 14.1±7.8 2) 23.1±6.9 2) | 34.2±8.8 22.9±6.9 2) 7.4±5.0 2) 10.3±5.3 2) 5.9±4.8 2) | / 33.0 78.4 69.9 82.7 |
Annotate: compare with model control group,
1)P＜0.05,
2)P＜0.01.
The result shows that Radix Angelicae Sinensis polysaccharide of the present invention can obviously produce the pain caused provide protection of Dichlorodiphenyl Acetate only for Radix Angelicae Sinensis Crude polysaccharides 1/10 dosage the time, and strengthens with dosage increase effect.
(2) Radix Angelicae Sinensis polysaccharide of the present invention causes the influence experiment of pain effect to hot plate method.The results are shown in Table 2.
Table 2 Radix Angelicae Sinensis polysaccharide of the present invention influences .n (routine number)=10, x ± s (mean ± standard deviation) to what the female mice hot plate method caused pain
| Grouping | Dosage (gkg -1) | The administration preceding threshold of pain time (s) | The threshold of pain time (s) after the administration | The time (s) is improved in the threshold of pain | Threshold of pain raising rate (%) |
| A kind of Radix Angelicae Sinensis polysaccharide of model group Radix Angelicae Sinensis Crude polysaccharides | / 0.075 0.250 0.025 0.075 | 20.8±6.4 21.5±6.3 23.4±4.4 20.7±5.2 21.3±6.2 | 21.2±6.8 24.9±7.7 34.6±9.3 29.4±9.1 34.8±8.7 | 1.3±1.5 4.4±5.4 11.7±8.2 1) 9.1±8.1 1) 13.7±9.6 1) | 1.9 15.8 47.9 42.0 63.4 |
Annotate: compare with control group,
1)P＜0.01.
The result shows that Radix Angelicae Sinensis polysaccharide of the present invention can significantly protect thermal stimulus pain caused, and strengthens with dosage increase effect only for Radix Angelicae Sinensis Crude polysaccharides 1/10 dosage the time.
(3) Radix Angelicae Sinensis polysaccharide of the present invention is to " dysmenorrhea model "---and stilboestrol and oxytocin bring out the influence experiment of Mouse Uterus spasm, the results are shown in Table 3.
Table 3 Radix Angelicae Sinensis polysaccharide of the present invention is to the influence of stilboestrol and oxytocin induced mice hysterospasm.
N (routine number)=10, x ± s (mean ± standard deviation)
| Grouping | Dosage (gkg -1) | Writhing response | ||
| Latent period (min) | Number of times (inferior/30min) | Incidence (%) | ||
| A kind of Radix Angelicae Sinensis polysaccharide of model group Radix Angelicae Sinensis Crude polysaccharides | / 0.075 0.250 0.025 0.075 | 12.3±3.8 21.1±9.0 1) 23.2±8.8 2) 25.4±6.6 2) 27.7±4.9 2) | 6.3±4.0 2.0±2.1 1) 1.3±1.9 2) 0.9±1.3 2) 0.6±1.3 2) | 100 60 40 1) 40 1) 30 2) |
Annotate: compare with model group,
1)P＜0.05,
2)P＜0.01.
The above results shows, disposable oral administration 0.025g/kg of Radix Angelicae Sinensis polysaccharide of the present invention and 0.075g/kg cause that to chemical substance and thermal stimulus the model of pain has obvious analgesic activity, and obviously suppress " dysmenorrhoea " induced mice writhing response and can effectively resist female hormone and bring out " dysmenorrhoea " caused pain reaction.Description of test, Radix Angelicae Sinensis polysaccharide of the present invention promptly has obvious effect under Radix Angelicae Sinensis Crude polysaccharides 1/10 dosage, and strengthens with dosage increase effect.
Claims (4)
1. Radix Angelicae Sinensis polysaccharide is made by laxative remedy: the angelol hypostasis is dissolved in distilled water, does not have the protein absorption peak of 280nm, the centrifugal precipitation of removing with Sevag method Deproteinization until ultraviolet detection; Keep water, add dehydrated alcohol, the precipitation polysaccharide is abandoned supernatant liquor, the gained throw out is with absolute ethanol washing, and after lyophilize, the gel molecular exclusion chromatography separates, and collects the relative molecular mass scope 1,0000-11, the separated product between 0000 obtains Radix Angelicae Sinensis polysaccharide again after lyophilize; Described angelol hypostasis is pressed method and is made, and gets angelica sinensis, and the decocting that adds 5～7 times of angelica sinensis weight boiled 30～60 minutes, filters; The water that the gained dregs of a decoction is added 3～6 times of weight decocted 30～50 minutes, filtered; The decocting that the gained dregs of a decoction add 3～6 times of weight boiled 30～50 minutes, filtered; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 40%～60%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate once more, adding ethanol to ethanol content is 70%～80%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate for the third time, adding ethanol to ethanol content is 80%～90%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Merge three throw outs and get Radix Angelicae Sinensis alcohol precipitation thing.
2. the preparation method of the described Radix Angelicae Sinensis polysaccharide of claim 1 is characterized in that: the angelol hypostasis is dissolved in distilled water, does not have the protein absorption peak of 280nm, the centrifugal precipitation of removing with Sevag method Deproteinization until ultraviolet detection; Keep water, add dehydrated alcohol, the precipitation polysaccharide is abandoned supernatant liquor, the gained throw out is colourless with absolute ethanol washing to washings, discard washings, the throw out after the washing is through lyophilize, and the gel molecular exclusion chromatography separates, collect the relative molecular mass scope 1,0000-11, the separated product between 0000 obtains Radix Angelicae Sinensis polysaccharide again after lyophilize; Described angelol hypostasis is pressed method and is made, and gets angelica sinensis, and the decocting that adds 5～7 times of angelica sinensis weight boiled 30～60 minutes, filters; The water that the gained dregs of a decoction is added 3～6 times of weight decocted 30～50 minutes, filtered; The decocting that the gained dregs of a decoction add 3～6 times of weight boiled 30～50 minutes, filtered; Merge three times filtrate, and filtrate is concentrated, be cooled to room temperature, adding ethanol to ethanol content is 40%～60%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate once more, adding ethanol to ethanol content is 70%～80%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Concentrate for the third time, adding ethanol to ethanol content is 80%～90%, 2～4 ℃ of refrigerations 24～48 hours, filters, and gets throw out; Merge three throw outs and get Radix Angelicae Sinensis alcohol precipitation thing.
3. the described Radix Angelicae Sinensis polysaccharide of claim 1 is in the purposes of preparation in the analgesic.
4. the described Radix Angelicae Sinensis polysaccharide of claim 1 is preparing the purposes for the treatment of in the dysmenorrhoea medicine.
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| CN101235097B (en) * | 2008-02-29 | 2010-11-24 | 武汉大学 | Method for extracting polysaccharide from angelica and application thereof |
| CN102154351B (en) * | 2010-12-30 | 2012-12-19 | 江苏大学 | Cationic angelica polysaccharide nanoparticle gene delivery system and preparation method thereof |
| CN106349401A (en) * | 2016-08-30 | 2017-01-25 | 甘肃中医药大学 | Method for optimizing parameters of radix angelica sinensis polysaccharide extracting technology based on response surface method |
| CN108771005A (en) * | 2018-06-19 | 2018-11-09 | 佛山推启农业研究院（普通合伙） | A kind of lactagogue forage and preparation method thereof |
| CN112870215A (en) * | 2018-09-26 | 2021-06-01 | 甘肃中医药大学 | Application of angelica polysaccharide |
| CN110283257A (en) * | 2019-06-28 | 2019-09-27 | 浙江理工大学 | A kind of extracting method of Radix Angelicae Sinensis polysaccharide |
| CN111297888A (en) * | 2020-04-01 | 2020-06-19 | 中国人民解放军第四军医大学 | Application of angelica polysaccharide in colitis |
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2004
- 2004-11-18 CN CN 200410061139 patent/CN1273493C/en not_active Expired - Fee Related
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