CN102166294B - Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof - Google Patents
Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof Download PDFInfo
- Publication number
- CN102166294B CN102166294B CN 201110087706 CN201110087706A CN102166294B CN 102166294 B CN102166294 B CN 102166294B CN 201110087706 CN201110087706 CN 201110087706 CN 201110087706 A CN201110087706 A CN 201110087706A CN 102166294 B CN102166294 B CN 102166294B
- Authority
- CN
- China
- Prior art keywords
- precipitation
- extract
- rhizoma anemarrhenae
- ethanol
- washing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a common anemarrhena polysaccharide extractive with immunosuppression activity and a preparation method and medicine purposes thereof. The preparation method comprising the steps of degreasing raw materials, extracting the degreased raw materials with water, precipitating the extracted material with alcohol, carrying out dialysis and ion-exchange column chromatography on the crude polysaccharide to eliminate proteins, pigments and molecular chemical components to produce an anemarrhena extractive. A huge amount of pharmacological experiments prove that common anemarrhena polysaccharide has an immunodepression activity and has the medical purposes of treating proinflammatory traumatic diseases, such as nephritis, pneumonia, upper respiratory tract infection, hepatitis, tonsillitis and diabetes.
Description
Technical field
The present invention relates to a kind of Rhizoma Anemarrhenae extract with immunosuppressive activity and preparation method thereof, and with this Rhizoma Anemarrhenae extract as immunosuppressive activity, be used for the treatment of the immunoinflammatory injury disease: the medical usage of nephritis, pneumonia, upper respiratory tract infection, hepatitis, tonsillitis and diabetes.
Background technology
The Rhizoma Anemarrhenae is the rhizome of the Liliaceae Anemarrhena plant Rhizoma Anemarrhenae (Anemarrhena asphodeloides Bge.), and nature and flavor are bitter, sweet, cold, return lung, stomach, kidney channel.Have clearing away heat-fire, nourshing Yin and drynsessmoistening prescription function, be used for fever caused by exogenous pathogenic factors, high hot excessive thirst, lung-heat type cough, osteopyrexia and fever, interior-heat is quenched one's thirst, the dryness of the intestine constipation.Chemical constituent in the modern study discovery Rhizoma Anemarrhenae is take steroidal saponin, two benzene pyrrones as main, lignanoids, flavonoid, organic acid etc. are still arranged, it is one of focus medicine in the modern medicine research, progress in recent years is rapid, but mostly concentrate on timosaponin class, the two benzene pyrrones composition, think that timosaponin is the important effective substance of the Rhizoma Anemarrhenae, other compositions of the Rhizoma Anemarrhenae are comprised that polysaccharide researches is less, there is not yet the report that it has immunosuppressive activity.The present invention is by deep chemistry, pharmacology, pharmacodynamic study, invented a kind of preparation method with immunosuppressive activity Rhizoma Anemarrhenae extract, the method is simple to operate, economical and energy saving, and can obtain the anemaran constituents more than 60% that content accounts for extract weight percentage ratio; Simultaneously find that first this Rhizoma Anemarrhenae extract has immunosuppressive activity, can be used for treating the various diseases relevant with the immunoinflammatory damage: nephritis, pneumonia, upper respiratory tract infection, hepatitis, tonsillitis and diabetes.The research of saponins, two benzene pyrrones effective substances is diverse in this and the Rhizoma Anemarrhenae former studies.
Summary of the invention
One of the object of the invention provides a kind of Rhizoma Anemarrhenae extract with immunosuppressive activity.
Two of the object of the invention provides a kind of preparation method of above-mentioned Rhizoma Anemarrhenae extract, and wherein the content of anemaran accounts for more than 60% of extract weight percentage ratio.
Three of the object of the invention is above-mentioned Rhizoma Anemarrhenae extract to be made the immunosuppressant of various dosage forms, is used for the treatment of the immunoinflammatory injury disease: the medical usage of nephritis, pneumonia, upper respiratory tract infection, hepatitis, tonsillitis and diabetes.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A kind of method for preparing above-mentioned Rhizoma Anemarrhenae extract, it comprises following concrete steps:
(1) Rhizoma Anemarrhenae is removed oil-soluble impurities with the high concentration ethanol backflow; (2) residue adds the water reflux, extract,, and filtrate is concentrated, adds the ethanol precipitate with ethanol, centrifugal must the precipitation; (3) precipitation is used dehydrated alcohol successively, and acetone washs respectively; (4) the precipitation dissolved in distilled water after the washing, dialysis; (5) dialysis solution is concentrated, ethanol precipitate with ethanol, centrifugal must the precipitation; (6) precipitation is used dehydrated alcohol successively, and acetone washs respectively; (7) the precipitation dissolved in distilled water after the washing adopts weak acid and weak base type anion and cation exchange resin series process to slough pigment and protein, and the concentrating under reduced pressure eluent namely gets this Rhizoma Anemarrhenae extract.
In order to reach better extraction effect, preferred, in the step (1) with the Rhizoma Anemarrhenae with 80～95% alcohol reflux defats of 6～12 times of weight 2～3 times, 2～3h refluxes at every turn; The distilled water reflux, extract, 2～3 times that in the step (2) residue is added 6～12 times of weight, each reflux, extract, 2～3h is concentrated into 0.2～1.0 volume with filtrate, adds 80～95% ethanol precipitate with ethanol, centrifugal must the precipitation; To precipitate the dehydrated alcohol of using successively 2～4 times of weight in step (3), (6), acetone washs respectively; Precipitation in the step (4) is redissolved with the distilled water of 0.5～2.0 times of weight, take molecular size range as 3.0 * 10
3-1.2 * 10
4Bag filter dialysis 24～72h.
To from the Rhizoma Anemarrhenae, adopt successively DEAE-Sepharose F.F and DEAE-52 ion-exchange chromatography (eluting is the NaCl solution of 0.1-1mol/L mutually) by isolated polysaccharide, and collect liquid and can select but be not limited to following gelose gel column chromatography to do and be further purified separation: Sephadex G50, Sephacryl S100, Sephacryl 200, Sephacryl S300 and Sephacryl S400 (eluting is 0.1-1mol/L NaCl solution mutually).
Described Rhizoma Anemarrhenae extract has immunosuppressive activity: the mouse monokaryon macrophage phagocytic function is inhibitory action, and prompting has the Non-specific immune suppression function; The content of experimental rat Hemolysin formation due to the reduction sheep red blood cell (SRBC), expression has the humoral immunization inhibit feature.Rhizoma Anemarrhenae extract can be used for treating the immunoinflammatory injury disease: nephritis, pneumonia, upper respiratory tract infection, hepatitis, tonsillitis and diabetes by immunosuppression mechanism.
After Rhizoma Anemarrhenae extract of the present invention can add various adjuvants and pharmaceutically acceptable carrier, excipient or diluent required when preparing different dosage form, method of Chinese medicinal with routine is prepared into any suitable clinical preparation, such as being injection (powder pin, freeze-dried powder, liquid drugs injection, transfusion etc.), oral formulations (tablet, oral liquid, granule, capsule, soft capsule or drop pill) etc.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not consisted of any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can make amendment or replace the details of technical solution of the present invention and form, but these modifications and replacing all fall within the scope of protection of the present invention.
Embodiment 1
Get the about 2.8kg of rhizoma ane marrhenae, with 90% alcohol reflux 3 times, each 2h, filter, filtering residue is dried 10 times of amounts of residue adding distil water, reflux, extract, 3 times, each 2h filters, merging filtrate, decompression and solvent recovery be to 1 times of volume of medical material, transfers to 95% ethanol that to contain the alcohol amount be 80%, hold over night, 2000rpm/min is centrifugal, and precipitation with 4 times of amount dehydrated alcohol, washing with acetone, gets crude extract successively.Getting crude extract is dissolved in water, the supernatant flowing water dialysis 48h in the bag filter that packs into, distill water dialysis 24h, dialysis solution is evaporated to medical material 1 volume, add 95% ethanol and transfer that to contain the alcohol amount be 85%, hold over night, 2000rpm/min is centrifugal, precipitation is washed successively with 4 times of amount dehydrated alcohol, acetone, distilled water redissolves, slough pigment and protein lyophilization through weak acid and weak base type anion and cation exchange resin (Amberlite FPA90Cl+Amberlite IRC84), get refining Rhizoma Anemarrhenae extract.Use the phenolsulfuric acid method, at the 500nm place, determined by ultraviolet spectrophotometry total polysaccharides content is with glucose C
6H
12O
6Count 65%.
Embodiment 2
Get the about 3kg of rhizoma ane marrhenae, with 80% alcohol reflux 2 times, each 3h, filter, filtering residue is dried 8 times of amounts of residue adding distil water, reflux, extract, 2 times, each 3h filters, merging filtrate, decompression and solvent recovery be to 1 times of volume of medical material, transfers to 95% ethanol that to contain the alcohol amount be 80%, hold over night, 2000rpm/min is centrifugal, and precipitation with 3 times of amount dehydrated alcohol, washing with acetone, gets crude extract successively.Getting crude extract is dissolved in water, supernatant flowing water dialysis 48h, distill water dialysis 24h, dialysis solution are evaporated to medical material 1 volume, add 95% ethanol and transfer that to contain the alcohol amount be 80%, hold over night, 2000rpm/min is centrifugal, and precipitation is washed successively with 3 times of weight dehydrated alcohol, acetone, and distilled water redissolves, slough pigment and protein lyophilization through weak acid and weak base type anion and cation exchange resin (Amberlite FPA90Cl+Amberlite IRC84), get refining Rhizoma Anemarrhenae extract.Use the phenolsulfuric acid method, at the 500nm place, determined by ultraviolet spectrophotometry total polysaccharides content is with glucose C
6H
12O
6Count 60%.
Embodiment 3 makes infusion solution
Rhizoma Anemarrhenae crude extract is done and is further purified separation through agarose gel post Sephacryl S300, Sephadex G50 chromatograph: obtain the above anemaran extract of 98% purity, add an amount of solubilizing agent, grind, add again a small amount of water for injection and dilute mixing, then it is an amount of to add sodium chloride, inject water to again ormal weight after the dissolving, filter embedding, sterilization, and get final product.
Embodiment 4 makes tablet
It is an amount of to get Rhizoma Anemarrhenae extract, adds the right amount of auxiliary materials such as diluent, disintegrating agent, mixing, granulation, drying, compacting in flakes, coating or spray film-coat and get final product.
Experimental example 1 Rhizoma Anemarrhenae extract antiinflammatory experimentation
Get the ICR male mice, be divided into immediately Rhizoma Anemarrhenae high dose (440mg/kg), Rhizoma Anemarrhenae low dosage (110mg/kg), positive control dexamethasone (3.6mg/kg) and blank group.Weigh and respectively organize Mouse Weight, the computation of mean values standard deviation is determined without group difference, gastric infusion is three days continuously, the 3rd day 50min after the last administration, every Mus auris dextra are coated with proinflammatory agent (dimethylbenzene 20 μ l), and left ear is as blank, after 3 hours mice is put to death, cut two ears and lay round auricle, analytical balance is weighed, and calculates the mice ear degree.The diversity that compares 4 groups of swellings.Experimental result sees Table 1.
Table 1: Rhizoma Anemarrhenae extract antiinflammatory experimental result
Annotate: compare with the blank group
*P＜0.01
Experimental result: each administration group compares with the blank group, the left and right sides ear difference weight saving of Rhizoma Anemarrhenae high and low dose group mice, and it is relevant to be dosage, and high dose group has utmost point significant difference (p＜0.01).Show that Rhizoma Anemarrhenae extract can reduce the mice ear degree, has certain antiinflammatory action.
The research of experimental example 2 Rhizoma Anemarrhenae extract refrigeration functions
Get male 80 of Wistar rat, be divided at random Rhizoma Anemarrhenae high dose (308mg/kg), low dosage (77mg/kg), positive control Dexamethasone group (3.78mg/kg) and model control group by body weight.Use first the normal anus temperature of every Mus of anus temperature instrumentation amount (anus temperature meter inserts anal 1.5cm) secondary, the initial body temperature of mice respectively organized in record, ask its meansigma methods as normal body temperature, the computation of mean values standard deviation determines that without group difference fasting be can't help distinguishing gastric infusion and normal saline (model control group) behind the water 12h.Subcutaneous injection 2,4-DNP (30mg/kg) pyrogenicity behind the gastric infusion 30min is to 0.5 hour, 1 hour, 2 hours, 4 hours and the variation of 6 hour record rat temperatures behind the heating agent.The results are shown in Table 2.
Table 2: the analgesic experimental result of Rhizoma Anemarrhenae extract
Annotate: each group compares at same time with the blank group
*P＜0.05,
*P＜0.01
Experimental result: the normal body temperature of each treated animal obviously raises to each the treated animal body temperature of different time behind the heating agent without obvious diversity (P＞0.05).Behind the pyrogenicity 0.5h, each treated animal body temperature obviously raises except the positive drug Dexamethasone group; Behind the pyrogenicity 1h, each is organized body temperature and all raises without obvious difference; Behind pyrogenicity 2h, the 4h, positive controls, Rhizoma Anemarrhenae extract high and low dose treated animal body temperature obviously reduce; Behind the pyrogenicity 6h, each treated animal body temperature convergence is normal.Show that Rhizoma Anemarrhenae extract can suppress the rat temperature rising that 2,4-dinitrophenol causes, has refrigeration function.
Experimental example 3 Rhizoma Anemarrhenae extracts are on the impact of immunosuppressed mice mononuclear-macrophage phagocytic function
Get BALB/C male mice 80, be divided at random blank group, Dexamethasone group and the high, medium and low dosage group of the Rhizoma Anemarrhenae, 15 every group, blank group gives normal saline, and other are respectively organized administration and see the following form, all with the administration of gavage mode, successive administration 7d.After the last administration 24 hours, inject the india ink 0.1ml/10g of 4 times of 1% gelatin dilution through mouse tail vein, respectively at getting blood 20 μ l behind 2min and the 6min socket of the eye, and it is joined 0.1%Na
2CO
3Shake up among the solution 2ml, with 0.1%Na
2CO
3Do blank, at spectrophotometer 600nm wavelength place's photometry density value, OD
1The optical density value of expression 2min, OD
2The optical density value of expression 6min.Index K is cleaned up in calculating according to formula, and the K value gets phagocytic index α after body weight and the conversion of liver spleen weight.Clean up index K=lgOD
1-lgOD
2/ t
2-t
1, phagocytic index α=body weight/liver spleen weight *
3√ K.Experimental result sees Table 3.
Table 3: Rhizoma Anemarrhenae extract is on the impact of immunosuppressed mice mononuclear-macrophage phagocytic function
Annotate: each group compares at same time with the blank group
*P＜0.05,
*P＜0.01
The experimental result Dexamethasone group compares with blank group, cleans up index K value and phagocytic index α value and all obviously reduces, and utmost point significant difference (p＜0.01) is arranged; Rhizoma Anemarrhenae high dose compares with blank group, and K value and α value all descend, K value (p＜0.05), and there were significant differences, and α value (p＜0.01) has utmost point significant difference.Dosage and Rhizoma Anemarrhenae low dosage and blank the group relatively in the Rhizoma Anemarrhenae are cleaned up index K value and phagocytic index α value all descends, and the α value has utmost point significant difference (p＜0.01), and the immunosuppressant index of α value is better than Dexamethasone group.Experimental result shows that Rhizoma Anemarrhenae extract has the Non-specific immune suppression effect.
Experimental example 4 Rhizoma Anemarrhenae extracts generate the impact test of (colorimetry) on the mice hemolytic antibody
Kunming mouse is divided into 4 groups at random, i.e. blank group, model control group, Rhizoma Anemarrhenae extract low dose group and high dose group.Every day, lumbar injection was 1 time, continuous 14 days.Except the blank group, all the other are respectively organized every mice and carry out immunity to 8 medicine pneumoretroperitoneum injection sheep red blood cell (SRBC) suspensions 0.2ml/ (about 400,000,000 cells), with normal saline serum is pressed 1: 300 dilution proportion, with the mice serum 1.0ml after the dilution, sheep red blood cell (SRBC) 0.5ml, add in the test tube, add again the guinea pig serum 1.0ml through normal saline dilution in 1: 10, blank is with equal-volume physiologic saline for substitute mice serum, put test tube in 37 ℃ of water-bath 10min, take out test tube and place ice-water bath, with cessation reaction, centrifugal after the cooling.With centrifugal rear supernatant 1.0ml, Dou Shi liquid 3.0ml adds in the test tube, and static 10min behind the mixing measures the trap value at the 540nm place.Trap value when in a test tube, adding in addition 0.25ml sheep red blood cell (SRBC) and Dou Shi liquid 3.75ml mensuration sheep red blood cell (SRBC) HD50.
The half hemolysis value HC of sample
50Trap value during=absorption of sample degree value X serum diluting multiple/sheep red blood cell (SRBC) HD50.Statistical method: all data mean ± SD represent, relatively check with t between group.Test and the results are shown in Table 4.
Table 4: the impact that the mice hemolytic antibody generates
Annotate: compare with the blank group,
△ △P＜0.01; Compare with model control group,
*P＜0.05,
*P＜0.01
Experimental result: model group and blank group relatively have utmost point significant difference (P＜0.01), show the modeling success.Rhizoma Anemarrhenae extract low dosage and model group relatively have significant difference (P＜0.05), and Rhizoma Anemarrhenae extract high dose and model group relatively have utmost point significant difference (P＜0.01).Experimental result shows that Rhizoma Anemarrhenae extract has certain inhibitory action to Hemolysin formation due to the sheep red blood cell (SRBC), illustrates to have the humoral immunization inhibit feature.
Experimental example 5 Rhizoma Anemarrhenae extract extracts are to the drug action of chronic glomerulonephritis rat
Get the SD rat, be divided at random blank group, model control group, Rhizoma Anemarrhenae extract low dose group, high dose group, 10 every group.Get the 70mg rabbit igg and be dissolved in the 10ml normal saline, with 60ml Freunds incomplete adjuvant mixing, make Emulsion, except the blank group, every rat gives 1ml, and the multiple spot subcutaneous injection is pre-immunity.Pre-immune rear the 7th day and the 8th day, the homemade nephrolytic sera 1mL/ that the tail vein injection dilution is a times only injected 2 days continuously for pathogenic immune.The immunity of causing a disease detects urine protein rear every day, and after urine protein was positive, blank group, model control group gave normal saline, and Rhizoma Anemarrhenae extract low dose group, high dose group give respectively Rhizoma Anemarrhenae extract, and dosage sees Table 5.Once a day, equal 4 weeks of successive administration.Detect urine protein, total serum protein and albumin under the laboratory condition.Experimental result sees Table 5.
Table 5: Rhizoma Anemarrhenae extract is to Chronic Glomerulonephritis Rats 24h urine protein, total serum protein and albuminous experimental result
Annotate: compare with the blank group,
△P＜0.05,
△ △P＜0.01; Compare with model control group,
*P＜0.05,
*P＜0.01
Experiment conclusion: model group and blank group relatively all have utmost point significant difference (P＜0.01), show the modeling success.Rhizoma Anemarrhenae extract low dosage and high dose group and model group comparison 24h urine protein content significantly reduce, total serum protein and albumin all have significant difference (P＜0.01), show that Rhizoma Anemarrhenae extract has therapeutical effect to nephrolytic sera type glomerulonephritis rat.
Experimental example 6 Rhizoma Anemarrhenae extracts cause the impact of rat chronic hepatic injury on carbon tetrachloride
Get Wistar kind rat, be divided at random Normal group, model control group, positive drug control group (liver adds glad group), the high, medium and low dosage group of Rhizoma Anemarrhenae extract.Except Normal group, each group 30%CCL
4The soybean oil solution gavage, secondary weekly, continuous three months.Except Normal group and model control group, each organizes gastric infusion simultaneously: it is 400mg/kg that liver adds glad group of dosage, and the high, medium and low dosage group dosage in full side is 440mg/kg, 220mg/kg, 110mg/kg.Normal group, model control group give with the volume distilled water, successive administration three months, fasting is 24 hours after three months, animal is plucked eyeball get blood, the concentration of the glutamate pyruvate transaminase (ALT) in the detection blood and glutamic oxaloacetic transaminase, GOT (AST), total protein (TP), albumin (ALB).The results are shown in Table 2, table 3, statistical method adopts the t check.
Table 6 Rhizoma Anemarrhenae extract causes the impact of chronic hepatic injury ALT, AST on CCL4
Annotate: compare with model group
*P＜0.05,
*P＜0.01
Table 6 shows: 1. to Serum ALT, the high, medium and low dosage group of Rhizoma Anemarrhenae extract all reduces by CCL significantly
4Due to the rising of ALT, but high dose group is best (P＜0.01), is dose-effect relationship; 2. to serum AST, high, medium and low dosage group can reduce by CCL significantly
4Due to the rising of AST, high dose group is best.
Table 7 Rhizoma Anemarrhenae extract is to CCL
4Cause the impact of rat chronic hepatic injury TP, ALB
Annotate: compare with model group
*P＜0.05,
*P＜0.01
Table 7 shows: 1. concerning serum T P, compare the high, medium and low dosage group of the extract CCL that all can obviously raise with the model contrast
4The TP that causes reduces, but high dose is best; 2. concerning serum ALB, compare with model group, high, medium and low dosage group all has the trend of rising ALB, but no significant difference.More than the explanation Rhizoma Anemarrhenae extract has protective effect for chronic hepatic injury, can be used for the treatment of hepatitis.
Claims (4)
1. Rhizoma Anemarrhenae extract with immunosuppressive activity is characterized in that being prepared from by the following method:
With 80～95% alcohol refluxs of 6～12 times of weight 2～3 times, the 2～3h that at every turn refluxes removes oil-soluble impurities with the Rhizoma Anemarrhenae; Residue adds the distilled water reflux, extract, 2～3 times of 6～12 times of weight, and each reflux, extract, 2～3h is concentrated into 0.2～1.0 volume with filtrate, 80～95% ethanol precipitate with ethanol, centrifugal must the precipitation; Precipitation is used respectively the dehydrated alcohol of 2～4 times of weight, washing with acetone successively; Precipitation after the washing is with the dissolved in distilled water of 0.5～2.0 times of weight, take molecular size range as 3.0 * 10
3-1.2 * 10
4Bag filter dialysis 24～72h; Dialysis solution is concentrated, 80～95% ethanol precipitate with ethanol, centrifugal must the precipitation; Precipitation is used respectively dehydrated alcohol, washing with acetone successively; Precipitation dissolved in distilled water after the washing adopts weak acid and weak base type anion and cation exchange resin series process to slough pigment and protein, and the concentrating under reduced pressure eluent namely gets this Rhizoma Anemarrhenae extract.
2. the preparation method of Rhizoma Anemarrhenae extract as claimed in claim 1 is characterized in that may further comprise the steps:
(1) Rhizoma Anemarrhenae is removed oil-soluble impurities with the high concentration ethanol backflow;
(2) residue adds the water reflux, extract,, and filtrate is concentrated, adds the ethanol precipitate with ethanol, centrifugal must the precipitation;
(3) precipitation is used respectively dehydrated alcohol, washing with acetone successively;
(4) the precipitation dissolved in distilled water after the washing is dialysed;
(5) dialysis solution is concentrated, adds the ethanol precipitate with ethanol, centrifugal must the precipitation;
(6) precipitation is used respectively dehydrated alcohol, washing with acetone successively;
(7) the precipitation dissolved in distilled water after the washing adopts weak acid and weak base type anion and cation exchange resin series process to slough pigment and protein, and the concentrating under reduced pressure eluent namely gets this Rhizoma Anemarrhenae extract.
3. according to the preparation method of Rhizoma Anemarrhenae extract claimed in claim 2, it is characterized in that: in the step (1) with the Rhizoma Anemarrhenae with 80～95% alcohol reflux defats of 6～12 times of weight 2～3 times, 2～3h refluxes at every turn; The distilled water reflux, extract, 2～3 times that in the step (2) residue is added 6～12 times of weight, each reflux, extract, 2～3h is concentrated into 0.2～1.0 volume with filtrate, 80～95% ethanol precipitate with ethanol, centrifugal must the precipitation; Precipitation in the step (4) is with the dissolved in distilled water of 0.5～2.0 times of weight, take molecular size range as 3.0 * 10
3-1.2 * 10
4Bag filter dialysis 24～72h; Adopt 80～95% ethanol precipitate with ethanol in the step (5); To precipitate dehydrated alcohol, the washing with acetone of using successively respectively 2～4 times of weight in step (3), (6).
4. according to claim 1,2 described Rhizoma Anemarrhenae extracts with immunosuppressive activity, it is characterized in that: the crude drug that can be made into infusion solution, tablet or other immunosuppressive drugs of conduct preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110087706 CN102166294B (en) | 2011-04-08 | 2011-04-08 | Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110087706 CN102166294B (en) | 2011-04-08 | 2011-04-08 | Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102166294A CN102166294A (en) | 2011-08-31 |
| CN102166294B true CN102166294B (en) | 2013-04-17 |
Family
ID=44487737
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110087706 Active CN102166294B (en) | 2011-04-08 | 2011-04-08 | Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102166294B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191289B (en) * | 2013-04-08 | 2015-05-13 | 广州中医药大学 | Synchronous preparation method of four effective parts in medicine pair of common anemarrhena rhizome and amur corktree bark and application thereof |
| CN103479856A (en) * | 2013-09-22 | 2014-01-01 | 山东省医药工业研究所 | Rhizoma anemarrhenae chemical component cluster extracts and preparation process thereof |
| CN108892739A (en) * | 2018-08-13 | 2018-11-27 | 黑龙江八农垦大学 | A kind of method for extraction and purification of anemaran |
| CN110051683B (en) * | 2019-05-06 | 2021-07-27 | 广州中医药大学(广州中医药研究院) | Application of rhizoma anemarrhenae polysaccharide in preparing medicine for treating xerophthalmia |
| CN110540603B (en) * | 2019-08-30 | 2021-12-24 | 南开大学 | Rhizoma anemarrhenae polysaccharide, and preparation method, identification method and application thereof |
| CN112480277B (en) * | 2019-09-12 | 2022-02-11 | 中国农业大学 | Process for refining and processing pleurotus ostreatus hypoglycemic polysaccharide |
| CN112876577B (en) * | 2021-03-18 | 2022-02-22 | 华南理工大学 | Homogeneous rhizoma anemarrhenae polysaccharide and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229316A (en) * | 2008-01-31 | 2008-07-30 | 广东药学院 | Rhizoma anemarrhenae extrac and applications as type 2 diabetes-curing medicine thereof |
-
2011
- 2011-04-08 CN CN 201110087706 patent/CN102166294B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229316A (en) * | 2008-01-31 | 2008-07-30 | 广东药学院 | Rhizoma anemarrhenae extrac and applications as type 2 diabetes-curing medicine thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102166294A (en) | 2011-08-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102166294B (en) | Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof | |
| CN100548323C (en) | A kind of pharmaceutical composition of making by Herb Gynostemmae Pentaphylli, Radix Panacis Quinquefolii and the Radix Astragali | |
| CN101554409B (en) | Long pepper alkaloid and preparation method, preparation and application thereof | |
| CN102772499B (en) | Effective parts obtained by alcohol extraction of raw materials of Relingqing granules as well as preparation method and applications thereof | |
| CN102138981B (en) | Preparation method of plantago seed polysaccharide extractive and applications thereof | |
| CN110551230B (en) | Preparation method of astragalus polysaccharide | |
| CN102772500B (en) | Relingqing Polygonum capitatum Buch-Ham ex D.Don raw material extract with anti-inflammatory action | |
| CN102210737B (en) | Sweet potato leaf extract and preparation method and use thereof | |
| CN110343185B (en) | Wine-processed polygonatum polysaccharide and application thereof in regulating spleen deficiency and immune function | |
| CN107778376A (en) | A kind of preparation method and applications of radix tetrastigme polysaccharide | |
| CN102225153B (en) | Traditional Chinese medicine composition for treating systemic lupus erythematosus and preparation method thereof | |
| CN101348812A (en) | Lichenin, and preparation and use thereof | |
| CN102813747B (en) | Traditional Chinese medicine for treating systemic lupus erythematosus | |
| CN106349304A (en) | Method for preparing high-purity Urticaside and Urticalignans | |
| CN102872015A (en) | Stephanotis total alkaloid extract as well as preparation method and application thereof | |
| CN108210600A (en) | A kind of preparation method and applications of limonin extract | |
| CN102805767A (en) | Heat stranguria removal granule raw material polygonum capitatum extract with anti-gonococcus effect | |
| CN110680802B (en) | Tetrandrine injection and preparation method thereof | |
| CN101538297A (en) | Preparation method of high-purity monomer flavone and general flavone contained in capsella bursa-pastoris and application of general flavone | |
| CN102139002B (en) | Cortex phellodendri polysaccharide extract and preparation method and medical application thereof | |
| CN113956375A (en) | Prepared rehmannia root homogeneous polysaccharide and preparation method and anti-depression effect thereof | |
| CN112546085A (en) | Sambucus chinensis extract for treating gout and preparation method thereof | |
| CN101554382B (en) | New purpose of triterpene substance | |
| CN103599189A (en) | Zhenqi Fuzheng (Chinese character) injection preparation and preparation method thereof | |
| CN104623388A (en) | Traditional Chinese medicine composition for treating systemic lupus erythematosus and preparation method of traditional Chinese medicine composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |