CN101953850B - Preparation method and application of oyster shell extract - Google Patents
Preparation method and application of oyster shell extract Download PDFInfo
- Publication number
- CN101953850B CN101953850B CN2010102803618A CN201010280361A CN101953850B CN 101953850 B CN101953850 B CN 101953850B CN 2010102803618 A CN2010102803618 A CN 2010102803618A CN 201010280361 A CN201010280361 A CN 201010280361A CN 101953850 B CN101953850 B CN 101953850B
- Authority
- CN
- China
- Prior art keywords
- oyster shell
- extract
- concha ostreae
- shell extract
- cell proliferation
- 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
Abstract
The invention relates to a preparation method of an oyster shell extract. The method comprises the following steps of: adding water-containing solution of alcohol into oyster shell powder in a material to liquid ratio of 1g:2-20mL, performing reflux extraction and filtering or centrifugally separating liquid out; and concentrating the obtained liquid under reduced pressure, extracting with an organic solvent, separating an organic solvent layer out, concentrating under reduced pressure and drying to obtain the oyster shell extract. Tests by a lissamine rhodamine B method indicate that the prepared oyster shell extract has the function of inhibiting cell proliferation of a plurality of tumor cell strains; and tests by a methyl thiazolyl tetrazolium method indicate that the oyster shell extract has the function of inhibiting cell proliferation of human umbilical vein endothelial cells. Experiments indicate that the oyster shell extract can play a role in resisting tumors by inhibiting tumor cell proliferation and angiogenesis. Therefore, the oyster shell extract of the invention can be taken as a cell proliferation inhibitor, an angiogenesis inhibitor and an anti-tumor agent.
Description
Technical field
The present invention relates to a kind of method for preparing and the application in preparation inhibition of cell proliferation, angiogenesis inhibitor and antitumor drug thereof of Concha Ostreae extract.
Background technology
In the sea-farming kind, characteristics such as shellfish is low owing to input cost, per unit area yield height, the proportion that accounts for sea-farming becomes the important channel that coastal fisherman increases income up to more than 80%.Be main with oyster culture in the shellfish culture; But residual a large amount of Concha Ostreaes in the Concha Ostreae process of manufacture; Except that part pulverize that the back is sold as animal feed or the calcining white lime, be not utilized effectively and deal carefully with, normal processed person is poured directly in highway both sides, vacant lot without fixed duties or the discarded irrigation canals and ditches; Serious environment pollution has directly influenced town and country human settlements' the improvement and the construction in new rural area.
Concha Ostreae uses as Chinese medicine, and the beginning is stated from Shennong's Herbal, also records for all previous Pharmacopoeia of the People's Republic of China; Be common ocean Chinese medicine, have tranquillization with heavy prescription, the YANG hyperactivity suppressing nourishing YIN; Hard masses softening and resolving is restrained astringent or styptic treatment for spontaneous sweating effect, is used for the palpitation with fear insomnia; Vertigo and tinnitus, treatment of conditions such as tuberculous lymphadenitis ， mass in the abdomen mass in the abdomen.Chinese medicine Concha Ostreae main component is inorganic constituentss such as calcium carbonate, and contained weight ratio accounts for more than 90%, contains the organic principle less than 10% in addition, like protein, glycoprotein and several amino acids etc.At present more to the method for preparing of soluble protein in the Concha Ostreae and the applied research in orthopaedics, the method for preparing and the applied research in preparation inhibition of cell proliferation, angiogenesis inhibitor or antitumor drug thereof of Concha Ostreae extract are not seen that so far report is arranged.
Summary of the invention
The purpose of this invention is to provide a kind of method for preparing and the application in preparation inhibition of cell proliferation, angiogenesis inhibition and antitumor drug thereof of Concha Ostreae extract, to satisfy the demand of prior art.
A kind of method for preparing of Concha Ostreae extract is characterized in that taking by weighing earlier oyster shell powder, is that 1g: 2-20mL adds aqueous alcoholic solution by solid-liquid ratio again, and reflux, extract, is filtered or the centrifugal liquid of telling; Gained liquid is used organic solvent extraction after under reduced pressure, concentrating, and obtains organic solvent layer, and concentrating under reduced pressure gets Concha Ostreae extract after the drying.
The application of above-mentioned Concha Ostreae extract in the preparation inhibition of cell proliferation.
The application of above-mentioned Concha Ostreae extract in the preparation angiogenesis inhibitor.
The application of above-mentioned Concha Ostreae extract in the preparation antitumor drug.
The present invention adopts lissamine rhodamine B (SRB) method to test the cell inhibitory effect effect of the Concha Ostreae extract of preparation to various tumor cell strains, adopts tetramethyl azo azoles salt (MTT) method to test the cell inhibitory effect effect of Concha Ostreae extract to Human umbilical vein endothelial cells.Experiment shows that this Concha Ostreae extract can and suppress the angiogenesis mode through the inhibition tumor cell proliferation, thereby brings into play its antitumor action.So Concha Ostreae extract useful as inhibitors of cell proliferation of the present invention, angiogenesis inhibitor and antitumor agent.
The specific embodiment
When preparing Concha Ostreae extract of the present invention, taking by weighing oyster shell powder earlier, is that 1g: 2-20mL adds aqueous alcoholic solution by solid-liquid ratio, and reflux, extract, is filtered or the centrifugal liquid of telling; Gained liquid is used organic solvent extraction after under reduced pressure, concentrating, and obtains organic solvent layer, and concentrating under reduced pressure gets Concha Ostreae extract after the drying.
Alcohol described in the present invention is methanol or ethanol, and aqueous concentration expressed in percentage by volume is 0-50%.Described organic solvent is dichloromethane, chloroform, ethyl acetate, ether or acetone.
Embodiment 1
Take by weighing 200g Concha Ostreae powder, place reaction vessel, add the aqueous alcoholic solution of 1000mL 50% (concentration expressed in percentage by volume, down with), reflux, extract, 1.5h tells liquid after centrifugal; Continue to add the 50% aqueous alcoholic solution of 1000mL to the gained medicinal residues, reflux, extract, 1h tells liquid after centrifugal.Merge 2 extracting solution of above-mentioned gained, be evaporated to 100mL after, with isopyknic dichloromethane extraction 2 times, combined dichloromethane extract, in 40 ℃ of concentrating under reduced pressure evaporates to dryness, Concha Ostreae extract.
Adopt lissamine rhodamine B (SRB) method, test the proliferation inhibition activity of prepared Concha Ostreae extract tumor cell.With human hepatocellular carcinoma BEL-7402 cell, human cervical carcinoma Hela cell and mouse leukemia P388 cell; Containing the RPMI-1640 culture medium of 10% hyclone respectively; At 32 ℃ (P388 cells) or at 37 ℃ (BEL-7402 and Hela cells), feed successive transfer culture in the incubator of 5% carbon dioxide.The take the logarithm tumor cell of trophophase, using fresh RPMI-1640 culture medium to be mixed with density is every milliliter 2 * 10
5The cell suspension of individual cell is inoculated in 96 orifice plates by every hole 200 μ L, and every hole adds the sample or the blank solution of 2 μ L variable concentrations, cultivates 24 hours down for 37 ℃.Then, 4 ℃, 3000 rev/mins centrifugal 3 minutes, inhale and to remove supernatant.Every hole adds 20% trichloroacetic acid, 50 μ L, places 4 ℃ to fix 1 hour, water flushing 5 times, and air drying.Every hole adds the acetum 50 μ L of 0.4%SRB and left standstill 30 minutes in room temperature.After 1% aqueous acetic acid cleaning 4 times, every hole adds 150 μ L Tris (Tris) buffer (10mmol/L, pH 10.5), utilizes MD company to produce SPECTRA MAX Plus type ELIASA and measures optical density (OD) value of every hole at the 520nm place.Each concentration of sample all is provided with three holes in same 96 orifice plate, and other establishes three hole blanks.Get the average OD value in three holes, by formula IR%=(OD blank-OD sample)/OD blank * 100% calculates the cell proliferation inhibition rate (IR%) under each concentration.Tumor cell proliferation suppresses experiment and shows that the Concha Ostreae extract of being produced is to the IC of mouse leukemia P388 cell
50Value is 67.3 μ g/mL; Concha Ostreae extract is respectively 64.84 and 77.78% to human cervical carcinoma Hela cell and human hepatocellular carcinoma BEL-7402 cell's proliferation inhibition rate under 250 μ g/mL concentration.Above experiment confirm, the Concha Ostreae extract of being produced can pass through to suppress the cell proliferation mode to tumor cell, thereby brings into play its antitumor action.
Adopt tetramethyl azo azoles salt (MTT) method, it is active to the inhibition of angiogenesis to test prepared Concha Ostreae extract.With Human umbilical vein endothelial cells HUVEHUVEC cell, containing the RPMI-1640 culture medium of 10% hyclone, 37 ℃, feed successive transfer culture in the incubator of 5% carbon dioxide.The take the logarithm Human umbilical vein endothelial cells HUVEC of trophophase buries 96 orifice plates behind the counting, concentration is 5000/hole.48h is hatched in RPMI-1640 cultivation with containing 10% hyclone.Use the RPMI-1640 culture medium that contains 1% hyclone instead, dosing continues to hatch 48h.Every then hole adds the MTT solution 20 μ L of 5mg/mL, hatches 4h.Sucking-off MTT adds 150 μ L dimethyl sulfoxide and fully dissolves; ELIASA 490nm place measures absorbance.Endothelial cell proliferation suppresses experiment and shows, the Concha Ostreae extract of being produced is 25.86% to the proliferation inhibition rate of Human umbilical vein endothelial cells HUVEC under 250 μ g/mL concentration.Above experiment confirm, the Concha Ostreae extract of being produced can pass through to suppress the mode of angiogenesis to tumor cell, thereby brings into play its antitumor action.
Embodiment 2
Take by weighing 200g Concha Ostreae powder, place reaction vessel, with the methanol of 1200mL, reflux, extract, 1h filters and tells liquid; To the methanol of gained medicinal residues continuation adding 1000mL, reflux, extract, 0.5h filters and tells liquid; Add the methanol of 1000mL once more to the gained medicinal residues, reflux, extract, 0.5h filters and tells liquid.Merge 3 extracting solution of above-mentioned gained, behind 40 ℃ of concentrating under reduced pressure evaporates to dryness, shared 200mL acetone divides 2 extractions, merges acetone extract, in 40 ℃ of concentrating under reduced pressure evaporates to dryness, gets Concha Ostreae extract.Tumor cell proliferation suppresses experiment and shows, the Concha Ostreae extract of being produced is 81.32% to the proliferation inhibition rate of mouse leukemia P388 cell under 100 μ g/mL concentration.
Claims (3)
1. the method for preparing of a Concha Ostreae extract is characterized in that taking by weighing earlier oyster shell powder, is that 1g: 2-20mL adds aqueous alcoholic solution by solid-liquid ratio again, and reflux, extract, is filtered or the centrifugal liquid of telling; Gained liquid is used organic solvent extraction after under reduced pressure, concentrating, and obtains organic solvent layer, and concentrating under reduced pressure gets Concha Ostreae extract after the drying; Said alcohol is methanol or ethanol, and aqueous concentration expressed in percentage by volume is 0-50%, and said organic solvent is dichloromethane, chloroform, ethyl acetate, ether or acetone.
2. the application of the described Concha Ostreae extract of claim 1 in the preparation tumor cell proliferation inhibitor.
3. the application of the described Concha Ostreae extract of claim 1 in the preparation antitumor drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102803618A CN101953850B (en) | 2010-09-08 | 2010-09-08 | Preparation method and application of oyster shell extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102803618A CN101953850B (en) | 2010-09-08 | 2010-09-08 | Preparation method and application of oyster shell extract |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101953850A CN101953850A (en) | 2011-01-26 |
| CN101953850B true CN101953850B (en) | 2012-05-30 |
Family
ID=43481608
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010102803618A Active CN101953850B (en) | 2010-09-08 | 2010-09-08 | Preparation method and application of oyster shell extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101953850B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106309507A (en) * | 2016-08-17 | 2017-01-11 | 青岛海洋生物医药研究院股份有限公司 | Composition having immunoregulatory activity as well as preparation and application thereof |
| CN107875174A (en) * | 2017-11-26 | 2018-04-06 | 防城港市蓝瀚达科技有限公司 | A kind of preparation method of oyster shell extract |
| CN108371650A (en) * | 2018-04-03 | 2018-08-07 | 广州赛莱拉干细胞科技股份有限公司 | Have effects that the composition of pox-eliminating whitening and pox-eliminating whitening skin care item |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1660897A (en) * | 2004-12-16 | 2005-08-31 | 集美大学 | Method for extracting and purifying glycoprotein shuck |
-
2010
- 2010-09-08 CN CN2010102803618A patent/CN101953850B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1660897A (en) * | 2004-12-16 | 2005-08-31 | 集美大学 | Method for extracting and purifying glycoprotein shuck |
Non-Patent Citations (1)
| Title |
|---|
| 吴杰连，等.海洋贝类抗癌免疫活性的研究进展.《水利渔业》.2006,第26卷(第3期),第16-18页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101953850A (en) | 2011-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102125552A (en) | Use of piperlongumine derivatives in preparation of medicines for treating cancers and medicinal compositions thereof | |
| CN102146054A (en) | Piperlongumine derivatives and medicinal composition and application to preparation of medicament for inhibiting tumor growth thereof | |
| CN101190258A (en) | Total sesquiterpene lactone extract containing rich parthenolide and preparation method and application thereof | |
| CN101953850B (en) | Preparation method and application of oyster shell extract | |
| CN110668936B (en) | Method for ultrasonically extracting benzophenone compounds from aquilaria sinensis leaves | |
| CN101259124A (en) | Pharmaceutical use of wedelolactone and its derivative | |
| CN102952756B (en) | Marine fungi altemaria MNP801 and application thereof | |
| TWI504394B (en) | Method for preparing antofine | |
| CN102775375B (en) | Chromone compound, preparation method and application of chromone compound, anti-aids pharmaceutical composition prepared from chromone compound and preparation of anti-aids pharmaceutical composition | |
| CN101190259A (en) | Southern magnolia total lactone extract and preparation method and application thereof | |
| CN103845420A (en) | Rheum nobile extract and application thereof | |
| CN101468061B (en) | Extract of immature exocarp of Juglans mandshurica Maxim., preparation method and medical use | |
| CN102552243A (en) | Application of myricanol and/or myricanone in preparing antitumor drugs | |
| CN110551090B (en) | Method for extracting antitumor active ingredients in traditional Chinese medicine rhizoma cibotii by ultrasonic waves | |
| CN103288615A (en) | Monocyclic phloroglucinol compounds and pharmaceutical composition and application thereof | |
| CN104606215B (en) | A kind of medicine for suppressing Enterovirus 71 | |
| CN104523740B (en) | A kind of selaginella doederlleini polyoses extract preparation method and applications | |
| CN106309470A (en) | Application of sargassum fusiforme polysaccharide | |
| CN103316120B (en) | The purposes of Clausena excavata Burm.f. or its extract | |
| CN108929320B (en) | 3R-indolylmethyl-6R-oxazolidinone modified piperazine-2, 5-dione, synthesis, activity and application thereof | |
| CN101390919A (en) | Use of pepperweed seed extract in preparing anti-tumor medicine | |
| CN101390979B (en) | Raisin tree seed extract and preparation method and use thereof in preparing anti-tumor medicine | |
| CN101313931B (en) | Application of Heichou extract in preparing antineoplastic medicament | |
| CN105748547A (en) | Method for separating active components from saussurea involucrata cell culture through extraction | |
| CN104758333B (en) | Paniculate swallowwort C21 steroidal extracts and its production and use |
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 |