CN105030811A - Anti-tumor application of polysaccharide of fruits with yellow thorns - Google Patents
Anti-tumor application of polysaccharide of fruits with yellow thorns Download PDFInfo
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Abstract
The invention provides application of polysaccharide of fruits with yellow thorns on preparation of anti-tumor food, medicines or health-care products. An in-vitro anti-tumor experiment proves that the polysaccharide of the fruits with the yellow thorns has good anti-tumor activity on human gastric carcinoma cells, breast cancer cells and hepatoma carcinoma cells which are in a logarithmic phase, and particularly has outstanding inhibitory activity on the hepatoma carcinoma cells. The polysaccharide of the fruits with the yellow thorns can be used for anti-tumor food, health-care products or medicines.
Description
Technical field
The invention belongs to field of health care products, be specifically related to the new medical use of the real polysaccharide of yellow bur.
Technical background
Yellow thorn, Berberidaceae plant straight fringe Radix Berberidis Amurensis BerberisdasystachyaMaxim., be the same with Cortex Acanthopanacis Radicis, Fructus Hippophae (black thorn) for covering, hiding, the medicinal traditional wild acinus with eating of ethnic groups such as Uygur, be referred to as " Qinghai three is stung ".At present, for Cortex Acanthopanacis Radicis and Hei Ci, investigation and application is very thorough, and for Huang thorn, its composition Study and function application are still comparatively blank.
In " Chinese herbal medicine handbook is commonly used in Qinghai ", Cortex berberidis dasystachyae (i.e. root bark) is documented: Cortex berberidis dasystachyae is the peel of stem of the straight fringe Radix Berberidis Amurensis of dicotyledon medicine Berberidaceae plant.Its fruit medicine function have also been made concise and to the point description, and it has treatment stomachache, dyspepsia, abdominal distention, the functions such as dysentery.
Yet there are no yellow bur real polysaccharide antineoplastic research report.
Summary of the invention
The object of the present invention is to provide the novelty teabag of the real polysaccharide of yellow bur.
Particularly, the invention provides the real polysaccharide of yellow bur and prepare the purposes in antineoplastic food, medicine or health product.
Wherein, described tumor is optimum or malignant tumor.
Further, described tumor is gastric cancer, hepatocarcinoma or breast carcinoma.
Further, described tumor is hepatocarcinoma.
Wherein, the real polysaccharide of described yellow bur, be the extract of Berberidaceae plant straight fringe Radix Berberidis Amurensis BerberisdasystachyaMaxim. fruit, in extract, polysaccharide is main component.
Measure in the specific embodiment of the invention and show, in extract, polyoses content is 93 ± 2%w/w, this purity of polysaccharide is higher, biological activity test is carried out with the polysaccharide under this purity, its result can reflect the biological activity of the real polysaccharide of yellow bur really, efficiently avoid the interference that other compositions measure active polysaccharide.On the bioactive basis of known polysaccharide, even if reduce the content of polysaccharide in extract, as long as suitably regulate extract consumption, make polysaccharide reach corresponding onset concentration, same or analogous biological activity can have been given play to equally.Therefore, the present invention limits, and in extract, polyoses content should be 60 ~ 95%w/w.
Further, in extract, polyoses content is 80 ~ 95%w/w, is further 90 ~ 95%w/w.
Wherein, the real polysaccharide of described yellow bur is real in raw material with yellow bur, is prepared by decoction and alcohol sedimentation technique.
Further, in precipitate with ethanol step, ethanol final concentration (i.e. alcohol content) reaches 60 ~ 85%v/v, is further 70 ~ 80%v/v, is preferably 75 ± 2%v/v.
Further, described decoction and alcohol sedimentation technique concrete operations are as follows: after real for yellow bur defat, removing monosaccharide, extracting in water, Aqueous extracts decolouring, removing protein, dialysis, except after micromolecule, add ethanol (making ethanol final concentration reach 60 ~ 85%v/v), leave standstill, completely to be precipitated, get precipitation, dry, namely obtain the real polysaccharide of yellow bur.
Further, the micromolecule molecular weight of dialysis removing is at below 3500Da.
Further, defat concrete operations are as follows: after real for yellow bur pulverizing, sieve, adopt petroleum ether or ether to be Extraction solvent, the residue filter after extraction, volatilizes solvent, obtain the real residue of the yellow bur of defat.
Further, go the concrete operations of monosaccharide as follows: gained is the real residue of the yellow bur of defat, and extract with ethanol (70 ~ 85%v/v), the residue after extraction volatilizes solvent, obtain except the real residue of the yellow bur of monosaccharide; Preferred concentration of alcohol is 80%v/v.
Further, dialysis adds ethanol to alcohol content (i.e. ethanol final concentration) to 70 ~ 80%v/v except after micromolecule, is preferably 75 ± 2%v/v.
Through above-mentioned purification step, the purity of the real polysaccharide of yellow bur effectively can be improved.Certainly, if only need to prepare the real polysaccharide of the lower yellow bur of purity, that can also omit in other purification steps except precipitate with ethanol is one or more.
In addition, for the desolventing technology of polysaccharide, usually adopt ion exchange (as DEAE-cellulose), oxidizing process (as H
2o
2), absorption method (as active carbon) etc., in actual use can according to self-demand, select in conjunction with the yield of later stage polysaccharide and purity the discoloration method be applicable to, in the present invention's detailed description of the invention, select H
2o
2decolouring, but this is not limited to the present invention and can only uses this kind of decolouring mode.
For removing protein, common method has trichloroacetic acid method, hydrochloric acid method, Sevage method (also on the books is Sevag method) etc., in actual use can according to self-demand, the method of removing protein be applicable to is selected in conjunction with the yield of later stage polysaccharide and purity, select Sevage method in the present invention's detailed description of the invention, but this is not limited to the present invention and can only uses this kind of removing protein mode.
Prove through anticancer experiment in vitro, the real polysaccharide of yellow bur all has good anti-tumor activity to being in five kinds of cell human gastric cancer cells BGC-823s of exponential phase, breast cancer cell MCF-7 and MDA-MB-231, hepatocellular carcinoma H22 and SMMC-7721, especially very remarkable to the inhibit activities of hepatoma carcinoma cell, real for yellow bur polysaccharide can be applied to antineoplastic food, health product or medicine.
Detailed description of the invention
Embodiment 1: the preparation of the real polysaccharide of yellow bur
1) the real defat of yellow bur: the fruit 400g getting ripe Huang thorn (i.e. straight fringe Radix Berberidis Amurensis) is dried, grinding and sieving; Get yellow bur to mix with petroleum ether in fact, supersound extraction after mixing, centrifugal removing extracting solution, residue filter, volatilizes petroleum ether, obtains the yellow bur reality of grease removal residue;
2) yellow bur is real in monosaccharide: the real residue of the yellow bur of grease removal mixes with ethanol, and described concentration of alcohol is 70 ~ 85%v/v, reflux, extract, and centrifugal removing extracting solution, volatilizes ethanol after residue filter, obtains except the real residue of the yellow bur of monosaccharide;
3) yellow thorn crude polysaccharides extracts: by step 2) gained mixes with pure water except the real residue of the yellow bur of monosaccharide, microwave radiation exaraction, centrifugally obtains extracting solution, discard after residue filter; Extracting solution is concentrated obtains crude polysaccharides extracting solution;
4) purification of yellow thorn crude polysaccharides: adopt H successively in crude polysaccharides extracting solution
2o
2, carry out decolouring, Savege method except deproteinize, then dialysis removing below 3500Da small-molecule substance, obtains the polysaccharide Aqueous extracts of purification;
5) preparation of the real polysaccharide of yellow bur: by step 4) the polysaccharide Aqueous extracts concentrated solution of purification of gained adds alcoholic solution, and make ethanol final concentration reach 75%v/v, after the drying of gained precipitation, namely obtain the real Polysaccharide B DP of yellow bur and amount to 2.06g.
Embodiment 2: the preparation of the real polysaccharide of yellow bur
1) the real defat of yellow bur: the fruit 400g getting ripe Huang thorn (i.e. straight fringe Radix Berberidis Amurensis) is dried, grinding and sieving; Get yellow bur to mix with petroleum ether in fact, reflux, extract, centrifugal removing extracting solution, residue filter, volatilizes petroleum ether, obtains the real residue of the yellow bur of grease removal;
2) yellow bur is real in monosaccharide: the real residue of the yellow bur of grease removal mixes with ethanol, and described concentration of alcohol is 70 ~ 85%v/v, supersound extraction, and centrifugal removing extracting solution, volatilizes ethanol after residue filter, obtains except the real residue of the yellow bur of monosaccharide;
3) yellow thorn crude polysaccharides extracts: by step 2) gained mixes with pure water except the real residue of the yellow bur of monosaccharide, warm macerating extraction, centrifugally obtains extracting solution, discard after residue filter; Extracting solution is concentrated obtains crude polysaccharides extracting solution;
4) purification of yellow thorn crude polysaccharides: adopt H successively in crude polysaccharides extracting solution
2o
2, carry out decolouring, Savege method except deproteinize, then dialysis removing below 3500Da small-molecule substance, obtains the polysaccharide Aqueous extracts of purification; 5) preparation of the real polysaccharide of yellow bur: by step 4) the polysaccharide Aqueous extracts concentrated solution of purification of gained adds alcoholic solution, and make ethanol final concentration reach 80%v/v, gained precipitation is dry, namely obtains the real Polysaccharide B DP of yellow bur and amounts to 1.355g.
Embodiment 3: the preparation of the real polysaccharide of yellow bur
1) the real defat of yellow bur: the fruit 400g getting ripe Huang thorn (i.e. straight fringe Radix Berberidis Amurensis) is dried, grinding and sieving; Get that yellow bur is real mix with petroleum ether, reflux, extract, again after ultrasonic, centrifugal removing extracting solution, residue filter, volatilizes petroleum ether, grease removal yellow bur reality residue;
2) yellow bur is real in monosaccharide: the real residue of the yellow bur of grease removal mixes with ethanol, and described concentration of alcohol is 70 ~ 85%v/v, supersound extraction, and centrifugal removing extracting solution, volatilizes ethanol after residue filter, obtains except the real residue of the yellow bur of monosaccharide;
3) yellow thorn crude polysaccharides extracts: by step 2) gained mixes with pure water except the real residue of the yellow bur of monosaccharide, reflux, extract, centrifugally obtains extracting solution, discard after residue filter; Extracting solution is concentrated obtains crude polysaccharides extracting solution;
4) purification of yellow thorn crude polysaccharides: adopt H successively in crude polysaccharides extracting solution
2o
2, carry out decolouring, Savege method except deproteinize, then dialysis removing below 3500Da small-molecule substance, obtains the polysaccharide Aqueous extracts of purification;
5) preparation of the real polysaccharide of yellow bur: by step 4) the polysaccharide Aqueous extracts concentrated solution of purification of gained adds alcoholic solution, ethanol final concentration is made to reach 70%v/v, 20mL absolute ethanol washing 3 times of gained precipitation, precipitate after its washing carries out spraying dry, make moisture content of material be down to less than 10%, obtain the real Polysaccharide B DP of yellow bur and amount to 1.67g.
Carry out Anthrone-sulfuricacid method mensuration to the real polysaccharide of yellow bur prepared by the inventive method, wherein polyoses content is 93% ± 2w/w.
The real polysaccharide anticancer experiment in vitro of the yellow bur of embodiment 4
(1) Experimental agents: the real polysaccharide of yellow bur, for the present invention prepares gained, is white powder solid.The concentration of setting is 521 μ g/mL, 256 μ g/mL, 128 μ g/mL, 64 μ g/mL, 32 μ g/mL, 16 μ g/mL, 8 μ g/mL (the real polysaccharide dosage of the yellow bur of the present invention or concentration are all with polysaccharide Mass Calculation);
(2) reagent and instrument: DMEM culture medium, dimethyl sulfoxide DMSO, penicillin, streptomycin (Sigma Co., USA), calf serum (FCS) (U.S. Hyclone, SouthLogan, UT), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoles base (MTT) (U.S. Amresco0793); CO2 constant incubator; Bio-Rad680 type microplate reader, Bio Rad Laboratories; Five kinds of cell human gastric cancer cells BGC-823s of exponential phase, breast cancer cell MCF-7 and MDA-MB-231, hepatocellular carcinoma H22 and SMMC-7721 (life science institute cellular resources center, Shanghai).
(3) experimentation: get the cell of five kinds of exponential phases by 1*10
5cell density is inoculated in 96 orifice plates, overnight incubation, changes serum-free medium; Real for the yellow bur of preparing gained polysaccharide is diluted to variable concentrations (521 μ g/mL, 256 μ g/mL, 128 μ g/mL, 64 μ g/mL, 32 μ g/mL, 16 μ g/mL, 8 μ g/mL); Add in the cell of incubated overnight, set up blank group simultaneously; Jolt mix homogeneously, the CO2 constant incubator being placed in 37 DEG C of 5% concentration is cultivated, and after 48 hours, MTT (5mg/mL) 20 μ L is added in every hole, is placed in the CO of 37 DEG C of 5% concentration
2continue cultivation in constant incubator 4 hours, abandon supernatant (culture fluid), every hole adds the DMSO of 100 μ L, and sample jolts 15min, measures light absorption value (OD) by microplate reader in 570nm place; Experiment in triplicate.Cell inhibitory rate (CellVability) (%)=(OD matched group-OD experimental group)/(the blank group of OD matched group-OD) × 100%.
(4) anticancer experiment in vitro result
The real polysaccharide component of the yellow bur of table 1 is to the suppression ratio (%) of stomach cancer cell, hepatoma carcinoma cell, breast cancer cell
As shown in Table 1, the real polysaccharide variable concentrations of the yellow bur of the present invention acted on three class tumor cells after 48 hours, the growth of its tumor cell is all subject to suppression in various degree, and, along with the increase of the real polysaccharide concentration of yellow bur, inhibitory rate of cell growth also increases gradually, presents good dose-effect relationship, especially all reaches more than 40% to the suppression ratio of hepatoma carcinoma cell when 512 μ g/mL.Therefore, the real polysaccharide of yellow bur all has good anti-tumor activity to being in five kinds of cell human gastric cancer cells BGC-823s of exponential phase, breast cancer cell MCF-7 and MDA-MB-231, hepatocellular carcinoma H22 and SMMC-7721, is especially applicable to antagonism hepatoma carcinoma cell.
Claims (9)
1. the real polysaccharide of yellow bur is preparing the purposes in antineoplastic food, medicine or health product.
2. purposes according to claim 1, is characterized in that: described tumor is optimum or malignant tumor.
3. purposes according to claim 1 and 2, is characterized in that: described tumor is gastric cancer, hepatocarcinoma or breast carcinoma.
4. the purposes according to claims 1 to 3 any one, is characterized in that: described tumor is hepatocarcinoma.
5. the purposes according to Claims 1 to 4 any one, is characterized in that: the real polysaccharide of described yellow bur is the extract of Berberidaceae plant straight fringe Radix Berberidis Amurensis BerberisdasystachyaMaxim. fruit, and in extract, polyoses content is 60 ~ 95%w/w.
6. the real polysaccharide of yellow bur according to claim 5, it is characterized in that: in extract, polyoses content is 80 ~ 95%w/w, is 90 ~ 95%w/w further.
7. the purposes according to claim 1 ~ 6 any one, is characterized in that: described Huang perverse fruit polysaccharide is real in raw material with yellow bur, is prepared by decoction and alcohol sedimentation technique; Further, described decoction and alcohol sedimentation technique concrete operations are as follows: after real for yellow bur defat, removing monosaccharide, extracting in water, Aqueous extracts decolouring, removing protein, dialysis, except after micromolecule, add ethanol and reach 60 ~ 85%v/v to ethanol final concentration, leave standstill, completely to be precipitated, get precipitation, dry, namely obtain the real polysaccharide of yellow bur; Further, the micromolecule molecular weight of dialysis removing is at below 3500Da.
8. purposes according to claim 7, is characterized in that: defat concrete operations are as follows: after real for yellow bur pulverizing, sieve, adopt petroleum ether or ether to be Extraction solvent, the residue filter after extraction, volatilizes solvent, obtains the real residue of the yellow bur of defat; Go the concrete operations of monosaccharide as follows: gained is the real residue of the yellow bur of defat, and with 70 ~ 85%v/v ethanol extraction, the residue after extraction volatilizes solvent, obtain except the real residue of the yellow bur of monosaccharide; Preferred concentration of alcohol is 80%v/v.
9. purposes according to claim 7, is characterized in that: dialysis adds ethanol to alcohol content to 70 ~ 80%v/v except after micromolecule, is preferably 75 ± 2%v/v.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86103972A (en) * | 1986-06-06 | 1987-12-16 | 中国科学院西北高原生物研究所 | Cosmetic mede from sand bramble |
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2015
- 2015-06-09 CN CN201510323183.5A patent/CN105030811B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86103972A (en) * | 1986-06-06 | 1987-12-16 | 中国科学院西北高原生物研究所 | Cosmetic mede from sand bramble |
Non-Patent Citations (3)
| Title |
|---|
| E.G.MARTYNOV等: "POLYSACCHARIDES OF Berberis vulgaris", 《CHEMISTRY OF NATURAL COMPOUNDS》 * |
| 张华峰等: "5 种小檗科植物多糖含量及抗氧化活性分析", 《精细化工》 * |
| 高巍等: "生物多糖非免疫活性的抗癌作用机制", 《天津药学》 * |
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