CN102936291A - Method for preparing selenylation poly mannuronic acid and application of selenylation poly mannuronic acid - Google Patents
Method for preparing selenylation poly mannuronic acid and application of selenylation poly mannuronic acid Download PDFInfo
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- polymannuronic acid
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- sulfonation
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- AEMOLEFTQBMNLQ-VANFPWTGSA-N D-mannopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H]1O AEMOLEFTQBMNLQ-VANFPWTGSA-N 0.000 title abstract description 12
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 title abstract description 11
- 238000000034 method Methods 0.000 title abstract description 11
- 239000002253 acid Substances 0.000 claims abstract description 103
- 238000000502 dialysis Methods 0.000 claims abstract description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 208000024827 Alzheimer disease Diseases 0.000 claims abstract description 21
- 239000003513 alkali Substances 0.000 claims abstract description 20
- 229940082569 selenite Drugs 0.000 claims abstract description 11
- 239000003960 organic solvent Substances 0.000 claims abstract description 10
- 239000006228 supernatant Substances 0.000 claims abstract description 10
- 238000006555 catalytic reaction Methods 0.000 claims abstract description 9
- 238000005119 centrifugation Methods 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000007795 chemical reaction product Substances 0.000 claims abstract description 8
- MCAHWIHFGHIESP-UHFFFAOYSA-L selenite(2-) Chemical class [O-][Se]([O-])=O MCAHWIHFGHIESP-UHFFFAOYSA-L 0.000 claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 4
- 238000007710 freezing Methods 0.000 claims abstract description 4
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- 238000006277 sulfonation reaction Methods 0.000 claims description 37
- 238000002360 preparation method Methods 0.000 claims description 31
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 21
- 230000007935 neutral effect Effects 0.000 claims description 15
- AKEJUJNQAAGONA-UHFFFAOYSA-N sulfur trioxide Chemical compound O=S(=O)=O AKEJUJNQAAGONA-UHFFFAOYSA-N 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 238000013019 agitation Methods 0.000 claims description 7
- JUJWROOIHBZHMG-UHFFFAOYSA-N pyridine Substances C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 7
- 230000001105 regulatory effect Effects 0.000 claims description 7
- NGNBDVOYPDDBFK-UHFFFAOYSA-N 2-[2,4-di(pentan-2-yl)phenoxy]acetyl chloride Chemical class CCCC(C)C1=CC=C(OCC(Cl)=O)C(C(C)CCC)=C1 NGNBDVOYPDDBFK-UHFFFAOYSA-N 0.000 claims description 5
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 claims description 5
- 229940045511 barium chloride Drugs 0.000 claims description 5
- 229910001626 barium chloride Inorganic materials 0.000 claims description 5
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- 102000011759 adducin Human genes 0.000 claims description 3
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- UDYFLDICVHJSOY-UHFFFAOYSA-N sulfur trioxide pyridine complex Chemical compound O=S(=O)=O.C1=CC=NC=C1 UDYFLDICVHJSOY-UHFFFAOYSA-N 0.000 claims description 2
- 230000001603 reducing effect Effects 0.000 claims 1
- 229920001282 polysaccharide Polymers 0.000 abstract description 19
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- -1 sulfur trioxide-pyridine compound Chemical class 0.000 abstract description 8
- 238000001035 drying Methods 0.000 abstract description 5
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 abstract description 4
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 3
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- 229910052711 selenium Inorganic materials 0.000 description 20
- 239000011669 selenium Substances 0.000 description 20
- 229940091258 selenium supplement Drugs 0.000 description 20
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 14
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- 239000011734 sodium Substances 0.000 description 11
- 230000000694 effects Effects 0.000 description 9
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 description 3
- 229940065287 selenium compound Drugs 0.000 description 3
- 229960001471 sodium selenite Drugs 0.000 description 3
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 2
- 108010090931 Proto-Oncogene Proteins c-bcl-2 Proteins 0.000 description 2
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- 229910052717 sulfur Inorganic materials 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- CRTGSPPMTACQBL-UHFFFAOYSA-N 2,3-dihydroxycyclopent-2-en-1-one Chemical compound OC1=C(O)C(=O)CC1 CRTGSPPMTACQBL-UHFFFAOYSA-N 0.000 description 1
- PMYDPQQPEAYXKD-UHFFFAOYSA-N 3-hydroxy-n-naphthalen-2-ylnaphthalene-2-carboxamide Chemical compound C1=CC=CC2=CC(NC(=O)C3=CC4=CC=CC=C4C=C3O)=CC=C21 PMYDPQQPEAYXKD-UHFFFAOYSA-N 0.000 description 1
- 102000000132 Alpha tubulin Human genes 0.000 description 1
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- AEMOLEFTQBMNLQ-BZINKQHNSA-N D-Guluronic Acid Chemical compound OC1O[C@H](C(O)=O)[C@H](O)[C@@H](O)[C@H]1O AEMOLEFTQBMNLQ-BZINKQHNSA-N 0.000 description 1
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- 102000008114 Selenoproteins Human genes 0.000 description 1
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- 108010087230 Sincalide Proteins 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- 229960004373 acetylcholine Drugs 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920001586 anionic polysaccharide Polymers 0.000 description 1
- 150000004836 anionic polysaccharides Chemical class 0.000 description 1
- 230000007131 anti Alzheimer effect Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
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- MCAHWIHFGHIESP-UHFFFAOYSA-N selenous acid Chemical compound O[Se](O)=O MCAHWIHFGHIESP-UHFFFAOYSA-N 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention relates to the technical field of selenylation polysaccharide and provides a method for preparing selenylation poly mannuronic acid. The method comprises steps of suspending poly mannuronic acid in an anhydrous organic solvent; adding a sulfur trioxide-pyridine compound dissolved by the anhydrous organic solvent with stirring; conducting reaction for 1-10 hours at the constant temperature from 20 DEG C to 100 DEG C; adding ethanol; centrifugalizing a reaction product; cleaning by using the ethanol; adding water and dissolving; adjusting the Potential of Hydrogen (PH) value to the neuter by using alkali; conducting dialysis; conducting filtering; conducting freezing and drying; obtaining sulfonated poly mannuronic acid; subjecting the sulfonated poly mannuronic acid and selenite to the reaction in the acid solution at the catalysis of Ba2+; adding sulphate; extracting supernatant fluid through centrifugation; adjusting the Potential of Hydrogen (PH) value to the neuter by using the alkali; conducting dialysis; conducting filtering; conducting freezing and drying; and obtaining the selenylation poly mannuronic acid. The invention also provides application of the selenylation poly mannuronic acid which is prepared through the method in medicines and health products for prevention and curing of Alzheimer's disease.
Description
Technical field
The invention belongs to selenizing polysaccharide technical field, be specifically related to a kind of selenizing polymannuronic acid preparation method and application thereof.
Background technology
Algin is a kind of water-soluble acid polysaccharide, mainly extracts to obtain from the cell walls of the brown algas such as bulk kelp, sargassun, sea-tangle, has unique structure and biological activity.On its structure by α-Isosorbide-5-Nitrae-L-guluronic acid (α-Isosorbide-5-Nitrae-L-guluronic acid, be abbreviated as G) and β-l, the straight chain anionic polysaccharide that 4-D-mannuronic acid (β-l, 4-D-mannuronicacid are abbreviated as M) is formed by connecting by the l-4 glycosidic link.Whole molecule is made of three kinds of fragments: polymannuronic acid fragment (Poly-mannuronate, PM), guluronic acid fragment (Poly-guluronate, PG) and polymannuronic acid-guluronic acid heterozygosis section (PMG block).Therefore, can isolate single polymannuronic acid by existing acid hydrolysis method.Be connected with the acid chemical combination of carboxyl on the special C5 of being of polymannuronic acid structure, and sulfur-bearing not on the structure, it carried out selenizing have certain difficulty, therefore adopt earlier sulfonation again the method for selenizing it is carried out selenizing.
The selenium element is the necessary element of vital movement, and its existence form has two kinds of inorganic selenium and organoselenium, and common inorganic selenium has Sodium Selenite and sodium selenate, and organoselenium mainly is seleno-protein and selenium polysaccharide.Selenium can consist of some oxidasic active centre, but the resistance of oxidation of enhancing body and to the resistivity of relative disease.This has illustrated the direct relation of selenium and HUMAN HEALTH.Inorganic selenium has cumulative toxicity and mutagenesis, dosage is difficult to control during use, and organoselenium toxicity is low, side effect is little, not only can bring into play better selenium, and more remarkable than inorganic selenium on challenge, therefore, inorganic selenium is combined with polysaccharide organic makes it to be converted into the organic selenium compounds selenium polysaccharide, the physiology of selenium and polysaccharide and pharmacological function are optimized.
Polysaccharide is arranged at present to the effect of alzheimer's disease (AD), but there is not yet selenium polysaccharide to the report of AD effect.
Summary of the invention
Technical problem to be solved by this invention is to overcome the defective of prior art, and a kind of selenizing polymannuronic acid preparation method and application thereof are provided.
The invention provides a kind of preparation method of selenizing polymannuronic acid, it comprises the steps:
Preparation sulfonation polymannuronic acid: polymannuronic acid is suspended in the anhydrous organic solvent, under agitation condition, add the sulphur trioxide of anhydrous organic solvent dissolving-pyridine mixture, under 20～100 ℃ of stationary temperatures, reacted 1～10 hour, add 95% ethanol of 2～4 times of volumes, the centrifugation reaction product, use again 95% washing with alcohol, be dissolved in water, regulate pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid;
Preparation selenizing polymannuronic acid: be 1:0.1～10 in mass ratio with described sulfonation polymannuronic acid and selenite in acidic solution, Ba
2+Catalysis under reacted 2～20 hours under 20～100 ℃ of stationary temperatures, sulfonation polymannuronic acid and bariumchloride mol ratio are 1:1～100, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix, filters, and lyophilize obtains described selenizing polymannuronic acid.
And, provide the prepared selenizing polymannuronic acid of preparation method of above-mentioned selenizing polymannuronic acid to prevent and the medicine for the treatment of and the application in the healthcare products at Alzheimer's disease.
The preparation method of selenizing polymannuronic acid of the present invention adopts the again preparation method of selenizing of earlier sulfonation, and it is low to have a cost, the advantage that productive rate is high.And effectively remove excessive selenite radical by dialysis method, also can dialyse simultaneously and remove other small molecules mineral compound and oligose.Simultaneously, the prepared selenizing polymannuronic acid of this preparation method has the effect of prevention and treatment Alzheimer's disease.
Description of drawings
Fig. 1 is the infrared spectrogram of polymannuronic acid (a), sulfonation polymannuronic acid (b) and the selenizing polymannuronic acid (c) of the embodiment of the invention;
Fig. 2 is Se-PM of the present invention and Na
2SeO
3, sulfonation polymannuronic acid, PM, different polymerization degree oligosaccharide mixture (MOM) to H
2O
2Cell survival rate graph of a relation behind the N2a-APP695-sw cytoprotective of damage;
Fig. 3 is selenizing polymannuronic acid, selenocarrageenan and Na
2SeO
3Impact on BACE1 in the N2a-APP695-sw cell and APP protein expression: the former figure of Western blot detected result, the 1st, contrast, the 2nd, Se-PM, the 3rd, Se-Carr, the 4th, Na
2SeO
3
Fig. 4 is selenizing polymannuronic acid, selenocarrageenan and Na
2SeO
3Impact on BACE1 protein expression in the N2a-APP695-sw cell: Quantity one software carries out the result of photodensitometry to the BACE1 protein band, and the 1st, contrast, the 2nd, Se-PM, the 3rd, Se-Carr, the 4th, Na
2SeO
3
Fig. 5 is selenizing polymannuronic acid, selenocarrageenan and Na
2SeO
3Impact on APP protein expression in the N2a-APP695-sw cell: Quantity one software carries out the result of photodensitometry to the APP protein band, and the 1st, contrast, the 2nd, Se-PM, the 3rd, Se-Carr, the 4th, Na
2SeO
3
Embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, is not intended to limit the present invention.
The embodiment of the invention provides a kind of preparation method of selenizing polymannuronic acid, and it comprises the steps:
S01: preparation sulfonation polymannuronic acid: polymannuronic acid (PM) is suspended in the anhydrous organic solvent, under agitation condition, add the sulphur trioxide of anhydrous organic solvent dissolving-pyridine mixture, under 20～100 ℃ of stationary temperatures, reacted 1～10 hour, add 95% ethanol of 2～4 times of volumes, the centrifugation reaction product, use again 95% washing with alcohol, be dissolved in water, regulate pH to neutral with alkali, dialysis, filter, lyophilize gets sulfonation polymannuronic acid (S-PM);
S02: preparation selenizing polymannuronic acid: be 1:0.1～10 in mass ratio with described sulfonation polymannuronic acid and selenite in acidic solution, Ba
2+Catalysis under reacted 2～20 hours under 20～100 ℃ of stationary temperatures, sulfonation polymannuronic acid and bariumchloride mol ratio are 1:1～100, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix, filters, and lyophilize obtains described selenizing polymannuronic acid (Se-PM).
Among the step S01, polymannuronic acid is suspended in the anhydrous organic solvent, and preferably, described organic solvent is dimethyl formamide (DMF), and the mol ratio of described sulphur trioxide-pyridine mixture and PM is 100～1:1.Described reaction was preferably reacted 3～5 hours under 50～70 ℃ of stationary temperatures, added 95% alcohol of 2～4 times of volumes, and its purpose is termination reaction and alcohol precipitation polysaccharide.Centrifugation, with after the alcohol washing precipitation, will precipitate soluble in water, with alkali adjusting pH value to 7.Wherein, described alkali can be the highly basic such as NaOH, KOH.Dialysis filters, and lyophilize gets the sulfonation polymannuronic acid.
Step S02 is preferably, in mass ratio for 1:1～3 with described sulfonation polymannuronic acid and selenite in joining non-strong reductinic acid, such as dilute nitric acid solution, more preferably, the mass concentration of nitric acid is 0.5%, wherein, the mass concentration of sulfonation polymannuronic acid in acid solution is 0.1～0.3%, is preferably 0.2%.At Ba
2+Catalysis under, reacted 7～10 hours under 40～60 ℃ of stationary temperatures, sulfonation polymannuronic acid and bariumchloride mol ratio are 1:1～100, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix, filters, and lyophilize obtains described selenizing polymannuronic acid.Wherein, described lyophilize was specifically descended freezing 8～24 hours at-80 ℃, used the vacuum lyophilization freeze-drying again.
Prepared selenizing polymannuronic acid comprises the pyranose form sugar unit, i.e. β-l, and the 4-D-mannuronic acid, at least one hydroxyl hydrogen is replaced by selenous acid ester group or its salt on its C2, C4 and the C5 position.The infrared light collection of illustrative plates of described polymannuronic acid, sulfonation polymannuronic acid and selenizing polymannuronic acid as shown in Figure 1.From Fig. 1 (a), can find out, 939.99,1037.21 and 1082.53cm
-1Be the charateristic avsorption band of pyranose ring skeletal vibration, 820.40cm
-1Be the charateristic avsorption band of D-polymannuronic acid, prove to have D-polymannuronic acid sugar ring skeleton structure in this sample molecule.In the infrared light collection of illustrative plates of sulfonation polymannuronic acid (Fig. 1 b), 856.27 and 1255.80cm
-1Be respectively the stretching vibration peak of C-O-S and S=O, prove in this sample molecule to have sulfate group; 1418.67 and 1629.79cm
-1Be respectively symmetry and the asymmetric stretching vibration peak of COO-, prove in this sample structure to have carboxyl; 3442.25cm
-1Be the hydroxyl absorption peak, prove existence-OH in this sample structure.The IR spectrogram of contrast PM is found, has increased the absorption peak of sulfate group in the sulfonation polymannuronic acid, shows that sulfate group successfully is introduced in the molecular structure of PM (Fig. 1 c).Have been reported the infared spectrum of Sodium Selenite at 788.63cm
-1And 742.29cm
-1Two charateristic avsorption bands are arranged, and PM in this scope without absorption peak.In the infrared light collection of illustrative plates of Se-PM (Fig. 1 c), 3457.35cm-1 is the hydroxyl absorption peak, proves existence-OH in this sample structure; 1415.33 and 1614.22cm
-1Be respectively symmetry and the asymmetric stretching vibration peak of COO-, prove to have carboxy CO OH in this sample structure.The invariant position of each charateristic avsorption band, the basic configuration of polysaccharide remains unchanged, its charateristic avsorption band 828.76cm
-1Strength Changes is little, after the dialysis in the selenium polysaccharide without free selenite radical, confirm that polysaccharide is combined rear formation chemical bond with selenite radical, rather than adsorb.
And the embodiment of the invention provides the prepared selenizing polymannuronic acid of the preparation method of above-mentioned selenizing polymannuronic acid in Alzheimer's disease prevention and the medicine for the treatment of and the application in the healthcare products.
Alzheimer's disease is one group of primary degeneration brain degenerative disease that the cause of disease is not bright.A lot of diseases are in the senium, the onset of hiding, and the course of disease is slow and irreversible, clinically take the intelligence infringement as main.Pathological change is mainly the cortex diffuse atrophy, and ditch returns broadening, and the ventricles of the brain enlarge, and neurone reduces in a large number, and visible senile plaque (SP), neuroneme knot pathologies such as (NFT), and choline acetylase and acetyl choline content significantly reduce.Described selenizing polymannuronic acid is acted on the AD cell model, and research selenizing polymannuronic acid is to change and the mechanism of action of AD pathological index.
Oxidative stress participates in generation and the development of alzheimer's disease, and described selenizing polymannuronic acid (Se-PM) has good antioxidant effect.Be that the N2a-APP695-sw cell is by 5 * 10 with the AD cell model
3/ hole is inoculated in 96 porocyte culture plates, wash cell three times after cultivating 24h, the serum-free DMEM substratum that adds respectively many kinds of selenium compounds of 100 μ L and polysaccharide, the ultimate density that makes contained selenium are that 2.5 μ mol/L, polysaccharide are identical in quality with corresponding selenium polysaccharide, add 10 μ L behind the cultivation 24h again and contain H
2O
2Serum-free DMEM substratum, make H
2O
2Final concentration be 60 μ mol/L (n=6), continue to cultivate 24h, add CCK-8 reagent, detect cell survival rate.Fig. 2 is that selenium concentration is Se-PM and the Na of 2.5 μ mol/L
2SeO
3, and the oligosaccharide mixture (MOM) of sulfonation polymannuronic acid, PM and the different polymerization degree of 500 μ g/mL is avoided H to the N2a-APP695-sw cell
2O
2The provide protection result of oxidative damage (* * * p＜0.001VS contrast, ###p＜0.001VS H
20
2Model group).As can be seen from Figure 2, Se-PM, sulfonation polymannuronic acid, polymannuronic acid and Na
2SeO
3Can make H
2O
2The survival rate of the N2a-APP695-sw cell of processing significantly improves, and under the effect of MOM, cell survival rate does not have noticeable change.Therefore, Se-PM, sulfonation polymannuronic acid, PM and Na
2SeO
3Can protect the N2a-APP695-sw cell to avoid H
2O
2Oxidative damage.And this experiment AD model cell mitochondrial membrane potential after the Se-PM effect significantly increases, the release of cytochrome C reduces, the ROS level obviously reduces, the Bcl-2 protein expression obviously increases.Therefore, Se-PM can be by the generation of ROS in protective wire plastochondria integrity and biological function, the inhibition cell, thereby improves the Bcl-2 protein expression, and Promote cell's growth and propagation stop or delay the apoptosis of cell.
The selenizing polymannuronic acid is processed the AD cell to be studied.The AD cell model that adopts is the N2a-APP695-sw cell, is the Neuro-2a cell that has surely turned APP695 Swedish mutator gene.The vegetative period N2a-APP-sw cell dissociation of taking the logarithm becomes single cell suspension, regulates cell concn, it is inoculated on the culture dish, and 37 ℃, 5% CO
2Cultivate 24h in the incubator.Discard old nutrient solution, repeat to wash 3 times with serum free medium, add respectively the serum-free DMEM substratum of selenium compound to be studied and polysaccharide again, the final concentration that makes selenium is that 2.5 μ mol/L(polysaccharide are identical in quality with corresponding selenium polysaccharide), continue to cultivate 24h.Collecting cell adds cell pyrolysis liquid (RIPA), the ice bath lysing cell, and 4 ℃ in the centrifugal 1h of 14000rpm.Collect supernatant, in-80 ℃ of refrigerators, save backup.Adopt the BCA method to measure protein concn.By the variation of Western blot method detection with the important associated protein of AD, detect respectively corresponding albumen with rabbit source BACE1 antibody, mouse source APP antibody.Study the selenizing polymannuronic acid to the action effect of AD by research with important protein B ACE1 the relevant APP of AD.BACE1 is beta-secretase 1, and β-actin is beta-actin, and APP is the precursor protein of A β, and α-tublin is alpha-tubulin.The prepared Se-PM of the preparation method of above-mentioned selenizing polymannuronic acid processes AD model cell leach protein after N2a-APP695-sw24 hour, carrying out Western blot after BCA is quantitative analyzes, result such as Fig. 3～shown in Figure 5, Se-PM can highly significant reduction BACE1 protein expression (* * * P＜0.001), also can significantly reduce the expression (* P＜0.05) of APP.Its action effect significantly is better than selenocarrageenan (Se-Carr) and Sodium Selenite (Na
2SeO
3).This explanation Se-PM might prevent the formation of the important pathological characters beta amyloid of AD patch, therefore has the effect of anti-AD.
Below in conjunction with specific embodiment specific implementation of the present invention is described in detail.
Embodiment 1:
With 100mg after drying polymannuronic acid (PM) be suspended among the DFM, under agitation condition, add the 600mg sulphur trioxide of 5mL dry DMF dissolving-pyridine mixture, reaction is 4 hours under 60 ℃ of stationary temperatures, 95% the ethanol that adds 3 times of volumes, centrifugation reaction product, the washing with alcohol with 95%, be dissolved in water, regulate pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid, and wherein sulphur content is 12.5%, DS=1.29, average yield is 78%;
Preparation selenizing mannuronic acid: the HNO that described 100mg sulfonation polymannuronic acid and 200mg selenite is joined 50mL0.5%
3In the solution, 25mg BaCl
2The lower 60 ℃ of reaction 8h of catalysis, add vitriol to remove Ba
2+The centrifuging and taking supernatant liquor, regulate pH to neutral with alkali, again with the dialysis of the dialysis tubing of molecular weight cut-off 1000, detect till liquid does not redden in the dialysis tubing to xitix, filter, lyophilize, obtain described selenizing polymannuronic acid (Se-PM), record that selenium content is 437.25 μ g/g in the sample, selenizing process average productive rate is 70%.
Embodiment 2:
With 100mg after drying polymannuronic acid (PM) be suspended among the DFM, add the 600mg sulphur trioxide of 5mL dry DMF dissolving-pyridine mixture under agitation condition, reaction is 10 hours under 20 ℃ of stationary temperatures, adds 95% ethanol of 2 times of volumes, the centrifugation reaction product, washing with alcohol with 95% is dissolved in water, and regulates pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid;
Preparation selenizing mannuronic acid: the HNO that described 100mg sulfonation polymannuronic acid and 10mg selenite is joined 50mL0.5%
3In the solution, 25mg Ba (NO
3)
2The lower 20 ℃ of reaction 20h of catalysis, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix.Filter, lyophilize obtains described selenizing polymannuronic acid (Se-PM).
Embodiment 3:
With 100mg after drying polymannuronic acid (PM) be suspended among the DFM, add the 600mg sulphur trioxide of 5mL dry DMF dissolving-pyridine mixture under agitation condition, reaction is 1 hour under 100 ℃ of stationary temperatures, adds 95% ethanol of 4 times of volumes, the centrifugation reaction product, washing with alcohol with 95% is dissolved in water, and regulates pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid;
Preparation selenizing mannuronic acid: the HNO that described 100mg sulfonation polymannuronic acid and 1000mg selenous acid is joined 50mL0.5%
3In the solution, 1000mg BaCl
2The lower 100 ℃ of reaction 2h of catalysis, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix.Filter, lyophilize obtains described selenizing polymannuronic acid (Se-PM).
The above only is preferred embodiment of the present invention, not in order to limiting the present invention, all any modifications of doing within the spirit and principles in the present invention, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (9)
1. the preparation method of a selenizing polymannuronic acid is characterized in that, comprises the steps:
Preparation sulfonation polymannuronic acid: polymannuronic acid is suspended in the anhydrous organic solvent, under agitation condition, add the sulphur trioxide of anhydrous organic solvent dissolving-pyridine mixture, under 20～100 ℃ of stationary temperatures, reacted 1～10 hour, add 95% ethanol of 2～4 times of volumes, the centrifugation reaction product, use again 95% washing with alcohol, be dissolved in water, regulate pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid;
Preparation selenizing polymannuronic acid: be 1:0.1～10 in mass ratio with described sulfonation polymannuronic acid and selenite in acidic solution, Ba
2+Catalysis under reacted 2～20 hours under 20～100 ℃ of stationary temperatures, described sulfonation polymannuronic acid and bariumchloride mol ratio are 1:1～100, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix, filters, and lyophilize obtains described selenizing polymannuronic acid.
2. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, the mol ratio of described sulphur trioxide-pyridine mixture and polymannuronic acid is 100～1:1.
3. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, described alkali is highly basic.
4. the preparation method of selenizing polymannuronic acid as claimed in claim 1, it is characterized in that, described preparation sulfonation polymannuronic acid is: polymannuronic acid is suspended in the dimethyl formamide, the sulphur trioxide of adding dimethyl formamide dissolving-pyridine mixture under agitation condition, under 50～70 ℃ of stationary temperatures, reacted 3～5 hours, 95% the ethanol that adds 2～4 times of volumes, the centrifugation reaction product, washing with alcohol with 95% is dissolved in water, and regulates pH to neutral with alkali, dialysis, filter, lyophilize gets the sulfonation polymannuronic acid.
5. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, described preparation selenizing polymannuronic acid is: in mass ratio for 1:1～3 with described sulfonation polymannuronic acid and selenite in salpeter solution, Ba
2+Catalysis under reacted 7～10 hours under 40～60 ℃ of stationary temperatures, described sulfonation polymannuronic acid and bariumchloride mol ratio are 1:1～100, add vitriol to remove Ba
2+, the centrifuging and taking supernatant liquor is regulated pH to neutral with alkali, again with the dialysis tubing dialysis of molecular weight cut-off 1000, detects till liquid does not redden in the dialysis tubing to xitix, filters, and lyophilize obtains described selenizing polymannuronic acid.
6. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, described lyophilize is to descend freezing 8～24 hours at-80 ℃, uses the vacuum lyophilization freeze-drying again.
7. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, described acidic solution is non-strong reducing property acid solution.
8. the preparation method of selenizing polymannuronic acid as claimed in claim 1 is characterized in that, the mass concentration of described sulfonation polymannuronic acid in acid solution is 0.1～0.3%.
9. prevent and the medicine for the treatment of and the application in the healthcare products at Alzheimer's disease such as the prepared selenizing polymannuronic acid of the preparation method of the described selenizing polymannuronic acid of claim 1～8.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106581037A (en) * | 2016-12-28 | 2017-04-26 | 深圳大学 | Use of seleno-polymannuronic acid |
| WO2018119770A1 (en) * | 2016-12-28 | 2018-07-05 | 深圳大学 | Use of seleno-polymannuronate |
| CN109260210A (en) * | 2018-10-11 | 2019-01-25 | 深圳大学 | Oligomannuronic acid is inhibiting the application in total Tau expression, phosphorylation and aggregation |
| CN111303310A (en) * | 2020-03-12 | 2020-06-19 | 大连大学 | Preparation method of marine sulfuric acid galactohexaose and application of marine sulfuric acid galactohexaose in preparation of medicines for resisting Alzheimer disease |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2693198A1 (en) * | 1992-07-03 | 1994-01-07 | Univ Paris Curie | New selenium contg. glycosyl or (poly)saccharide derivs. - used e.g. as stereo-selective glycosylating agents, antiviral agents, enzyme inhibitors and their intermediates |
| CN1288899A (en) * | 2000-08-04 | 2001-03-28 | 国家海洋局第一海洋研究所 | Selenium compound of polysaccharide and its preparation |
| CN1562050A (en) * | 2004-03-24 | 2005-01-12 | 中国海洋大学 | Use of oligose alginate in anti-dementia and anti-diabetes |
| CN1600793A (en) * | 2004-09-06 | 2005-03-30 | 天津科技大学 | Polysaccharide of selenic acid and preparation method |
| CN101104643A (en) * | 2006-07-14 | 2008-01-16 | 陈艺新 | Polysaccharide sulfate selenide compound and its preparation method and application |
| JP2008260823A (en) * | 2007-04-11 | 2008-10-30 | Univ Of Fukui | Polysaccharide modified with selenious acid and its manufacturing method |
-
2012
- 2012-11-15 CN CN201210458914.3A patent/CN102936291B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2693198A1 (en) * | 1992-07-03 | 1994-01-07 | Univ Paris Curie | New selenium contg. glycosyl or (poly)saccharide derivs. - used e.g. as stereo-selective glycosylating agents, antiviral agents, enzyme inhibitors and their intermediates |
| CN1288899A (en) * | 2000-08-04 | 2001-03-28 | 国家海洋局第一海洋研究所 | Selenium compound of polysaccharide and its preparation |
| CN1562050A (en) * | 2004-03-24 | 2005-01-12 | 中国海洋大学 | Use of oligose alginate in anti-dementia and anti-diabetes |
| CN1600793A (en) * | 2004-09-06 | 2005-03-30 | 天津科技大学 | Polysaccharide of selenic acid and preparation method |
| CN101104643A (en) * | 2006-07-14 | 2008-01-16 | 陈艺新 | Polysaccharide sulfate selenide compound and its preparation method and application |
| JP2008260823A (en) * | 2007-04-11 | 2008-10-30 | Univ Of Fukui | Polysaccharide modified with selenious acid and its manufacturing method |
Non-Patent Citations (1)
| Title |
|---|
| YU CAO等: "Antioxidant activity and antitumor activity (in vitro) of xyloglucan selenious ester and surfated xyloglucan", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106581037A (en) * | 2016-12-28 | 2017-04-26 | 深圳大学 | Use of seleno-polymannuronic acid |
| WO2018119770A1 (en) * | 2016-12-28 | 2018-07-05 | 深圳大学 | Use of seleno-polymannuronate |
| CN106581037B (en) * | 2016-12-28 | 2019-09-17 | 深圳大学 | The application of selenizing polymannuronate |
| CN109260210A (en) * | 2018-10-11 | 2019-01-25 | 深圳大学 | Oligomannuronic acid is inhibiting the application in total Tau expression, phosphorylation and aggregation |
| CN111303310A (en) * | 2020-03-12 | 2020-06-19 | 大连大学 | Preparation method of marine sulfuric acid galactohexaose and application of marine sulfuric acid galactohexaose in preparation of medicines for resisting Alzheimer disease |
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