CN106309470A - Application of sargassum fusiforme polysaccharide - Google Patents
Application of sargassum fusiforme polysaccharide Download PDFInfo
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- CN106309470A CN106309470A CN201610763357.4A CN201610763357A CN106309470A CN 106309470 A CN106309470 A CN 106309470A CN 201610763357 A CN201610763357 A CN 201610763357A CN 106309470 A CN106309470 A CN 106309470A
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- sargassum fusiforme
- polysaccharide
- hijiki polysaccharide
- rsv
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses an application of sargassum fusiforme polysaccharide. The sargassum fusiforme polysaccharide has antiviral activity. The application of the sargassum fusiforme polysaccharide is the application of the sargassum fusiforme polysaccharide with antiviral activity in preparation of RSV (Respiratory Syncytial Virus) or EV71 (Human enterovirus 71) resistant drugs. The remarkable inhibition effect of the sargassum fusiforme polysaccharide on RSV and EV71 is discovered for the first time, and the virus TI (therapeutic index) of the drugs prepared from the sargassum fusiforme polysaccharide can be up to about 1000. The invention also discloses the application of the sargassum fusiforme polysaccharide in preparation of antiviral drugs, and the drugs can inhibit RSV and EV71 viruses. Therefore, the sargassum fusiforme polysaccharide can eliminate phlegm, soften hardness, induce diuresis, alleviate edema, eliminate heat and eliminate dampness and have potential application value in the field of treatment of RSV and EV71 virus infectious diseases.
Description
Technical field
The present invention relates to pharmaceutical technology field, be specifically related to Hijiki polysaccharide in preparation in suppression respiratory syncytial virus
(Respiratory Syncytial Virus, RSV), enterovirns type 71 (Human enterovirus 71, EV71) medicine
In application.
Background technology
Sargassum fusiforme (Harv.) Setch is Sargassaceae (Sargassceae) Sargassum plant Sargassum fusiforme (Harv.) Setch (Sargassum fusiforme
(Harv.) Setchell) dry frond.Sargassum fusiforme (Harv.) Setch is practised and is claimed " lobule Sargassum ", has another name called Ottelia acuminata (Gagnep.) Dandy, sharp, the extra large Fructus Hordei Vulgaris of Cornu Cervi etc..Main product
In Liaoning, Shandong, Fujian, Zhejiang, Deng Di marine site, Guangdong, mostly be wild, summer, season in autumn two fish, and its dry frond entirety is used as medicine.
It is listed under Compendium of Material Medica grass portion Sargassum item, records: " Sargassum fusiforme (Harv.) Setch salty energy relaxing the bowels with lubricant drugs, the cold drinking-water that can expel the heat-evil.Therefore can disappear goiter, tuberculosis,
And except edema, tinea pedis, prolonged fluid-retention, mental disorder damp and hot, make pathogen go out also from urine ".Begin to be loaded in Shennong's Herbal, this medicine is arranged
For middle product, call it and cure mainly " goiter stagnation of QI, dissipates stone pain, carbuncle lump in the abdomen heavily fortified point gas under neck, thunder up and down in abdomen, lower 12 edema ".
This product bitter in the mouth, salty, cold, nontoxic.Return liver, stomach, kidney channel.There is effect of eliminating phlegm and softening indurated mass, inducing diuresis to remove edema, dispelling heat and resolving dampness.Character identification:
All shrinkages dough in curling, pitchy, surface is by bloom, and length 15～40cm, matter is crisp frangible.Macerate rear meat stick-slip,
Pliable and tough, main shaft is cylindrical, rough surface, has brachyplast, branch alternate, and without thorn-like projection, cytofila is linear, club-like, slightly
Flat, tip expands hollow and becomes bubble sometimes, or becomes peltate.There are spindle or spherical air bag, length 5～10cm between axil, grow thickly, capsule
Handle is longer.Receptacle cylinder or oblong.Gas raw meat, salty in the mouth.
Hijiki polysaccharide (SPF) is to extract a kind of water soluble polysaccharide obtained from full algae Sargassum fusiforme (Harv.) Setch, Sargassum fusiforme (Harv.) Setch frond
Main component, is mainly made up of Algin and sulfated fucan.It is anti-that modern pharmacology research shows that Hijiki polysaccharide has
The effects such as tumor, blood sugar lowering, blood fat reducing, enhancing human body immunity power, slow down aging.The multiple pharmacologically active of Sargassum fusiforme (Harv.) Setch and its polysaccharide
Structure is closely related.SPF is the acidic polysaccharose mixture that a class composition structure is similar, and good water solubility, viscosity is high, mainly includes brown
Alginic acid (Acidic Polysaccharidesof Algae, APA), fucose (Fucoidan polysaccharide
Sulfate, FPS) and laminaran (Laminaran), in Sargassum fusiforme (Harv.) Setch the content analysis of polysaccharide be to Sargassum fusiforme (Harv.) Setch drug efficacy study and
The important evidence developed.
RSV is the most important virus causing disease of infant viral lower respiratory infection in world wide.Find the most again
Rsv infection is the important pathogen of immunosuppressant adult and old people.Overwhelming majority rsv infection is upper respiratory tract infection, but a lot
Baby can develop into pneumonia and lower respiratory infection.
EV71 is one of mankind's hand-foot-mouth disease (HFMD) main pathogens, is a class acute viral disease, has become
Worldwide public health problem.
Summary of the invention
It is desirable to provide the application of a kind of Hijiki polysaccharide.
For achieving the above object, the technical scheme is that
The application of a kind of Hijiki polysaccharide, described Hijiki polysaccharide has antiviral activity, answering of described Hijiki polysaccharide
With referring to that the Hijiki polysaccharide with antiviral activity resists for the application in RSV or EV71 virus drugs in preparation.
Preferably, Hijiki polysaccharide preparation method is decoction and alcohol sedimentation technique.
It is further preferred that the concretely comprising the following steps of described decoction and alcohol sedimentation technique, by Sargassum fusiforme (Harv.) Setch powder (45 DEG C of dry 2h) with steam
Decocting in water after distilled water mixing, change liquid and extract, after the filtrate after extracting concentrates, add 5% trichloroacetic acid, centrifugal segregation protein,
Then freezing after the solution dialysis that will obtain, adds 95% ethanol precipitate polysaccharides, and centrifuging and taking precipitates, and vacuum drying i.e. can get sheep
Dwell dish polysaccharide powder.
Further preferred, Sargassum fusiforme (Harv.) Setch powder is 1:30 with the mass ratio of distilled water.
Further preferred, changing liquid extraction time is 3 times.
Further preferred, centrifugal speed is 5000r/min, and centrifugation time is 20-30min.
Further preferred, dialysis time is 2.5-3.5 days.
The invention has the beneficial effects as follows:
Present invention firstly discovers that Hijiki polysaccharide all has significant inhibitory action to RSV and EV71 virus, its medicine presses down poison
Index (TI) up to about 1000, and disclose Hijiki polysaccharide be applied to prepare antiviral drugs, it is possible to suppression RSV, EV71
Virus, thus not only can eliminating phlegm and softening indurated mass, inducing diuresis to remove edema, dispelling heat and resolving dampness, and treatment RSV, EV71 disease of viral infection neck
There is potential using value in territory.
Accompanying drawing explanation
Fig. 1 is the microscope photograph of RSV virus control: complete CPE schematic diagram (200 ×) occurs in MA104 cell；
Fig. 2 is EV71 virus control microscope photograph: complete CPE schematic diagram occurs in RD cell；(200×)；
Fig. 3 is MA104 microcytoscope picture (200 ×) being protected；
Fig. 4 is RD microcytoscope picture (200 ×) being protected.
Detailed description of the invention
The invention will be further described below in conjunction with the accompanying drawings.
The Sargassum fusiforme (Harv.) Setch crude polysaccharides Effect study to virus:
(1) Hijiki polysaccharide crude extract is prepared:
Take Sargassum fusiforme (Harv.) Setch powder: distilled water=1:30 (g/g), decocting in water 4.5-6h, centre is changed liquid and extracted 3 times altogether.Take filtrate, rotation
After inspissation is condensed to the 1/4 of original volume, add 5% trichloroacetic acid adjusting pH is that 5.0-6.5,5000r/min are centrifuged 20min to remove
Protein in crude polysaccharides solution, gained solution loads in bag filter dialyses (about 3d), by solution freeze overnight, adds 4 times
95% ethanol precipitate polysaccharides of volume, centrifuging and taking precipitates, and vacuum drying i.e. can get crude polysaccharide powder end.By said extracted thing respectively
It is dissolved in cell maintenance medium (containing the 1640 of 2% new-born calf serum, GIBICO Products) standby.
(2) cell strain and Strain:
Thered is provided by microbial room of Basic Medical Science Inst., Shandong Prov. Academy of Medical Science: (rhesus embryonic kidney is thin for MA104
Born of the same parents, for RSV sensitive cells, as shown in Figure 3), (human rhabdomyosarcoma cells, for EV71 sensitive cells, as shown in Figure 4) for RD；EV71
(being isolatable from Jinan Infectious Disease Hospital hand-foot-mouth disease patient, turn out to be EV71 through complete sequence determination), and RSV (long strain, 2000 years
October is quoted from Virology Inst., China Academy of Preventive Medicine Sciences's seed culture of viruses room).
(3) cytotoxic assay:
Reference literature king little Yan, Zhang Meiying, Wang Yafeng etc., Semen Arecae extract In Vitro Anti Coxsackie B virus 3 and herpes simplex
Virus 1 type and the research of hepatitis B virus, time treasure's traditional Chinese medical science traditional Chinese medicines, 2008；The method of 19 (12): 2954～2955, o'clock 96
Cell on orifice plate grows up to monolayer, medicine cell maintenance medium starts from the 1st hole 12 concentration of 2 doubling dilution, is seeded in 96
On orifice plate, each dilution factor sets repetition 3 hole, at 37 DEG C, 5%CO2Under the conditions of cultivate, its CPE of basis of microscopic observation, continuous observation
Drug toxicity.CPE is considered as toxicity more than 50%.Then with neutral red staining, sets and measure wavelength as 540nm, survey its OD value, carefully
Born of the same parents' survival rate is experimental group result compared with cell controls group OD value, is calculated the poisoning of its half by Reed-Muench method
Concentration (toxic concentration 50%, TC50)。
TC50=[Antilog (log is higher than the value-pd of 50% pathological changes rate drug dilution degree)] × CFirst hole drug level。
(4) toxin inhibitory test:
Reference literature king little Yan, Zhang Meiying, Wang Yafeng etc., Semen Arecae extract In Vitro Anti Coxsackie B virus 3 and herpes simplex
Virus 1 type and the research of hepatitis B virus, time treasure's traditional Chinese medical science traditional Chinese medicines, 2008；The method of 19 (12): 2954～2955, o'clock 96
Cell on orifice plate grows up to monolayer, medicine cell maintenance medium starts from the 1st hole 12 concentration of 2 doubling dilution, is seeded in 96
On orifice plate (50 μ L/ hole), each dilution factor sets repetition 3 hole, if cell controls, virus control.In addition to cell controls, each hole adds
The virus 50 μ L of 100TCID50.35 DEG C, 5%CO2Under the conditions of cultivate, continuous observation cytopathy under microscope until virus right
Terminate after reaching 90% according to CPE cultivating.Measuring absorbance A value by microplate reader, it is effective that the dilution factor of CPE50% is considered as medicine half
Concentration (concentration for 50%of maximal effect, EC50)。
EC50=[Antilog (log is higher than the value-pd of 50% survival rate drug dilution degree)] × CFirst hole drug level.According to being surveyed
Medicine median toxic concentration (TC50) and medicine medium effective concentration (EC50), the two is compared acquisition and presses down poison index (TI), and TI is more than
4 are judged to effectively.
(5) result:
1. medicine median toxic concentration TC50Mensuration: examine under a microscope that cellular control unit form is good, adherent growth
Fine and close.Medicine group cell breakage, present poisoning form, record OD value, calculate the TC of medicine according to formula50.Hijiki polysaccharide
TC50It is 2 at MA104-1.3；It is 2 at RD-1.6。
2. Antiviral Effect test: basis of microscopic observation finds, the densification good, adherent of the cellular morphology of matched group；And
RSV, EV71 virus control group all about 24h occur the CPE, RSV of more than 90% in MA104 CPE with downright bad, be broken for main
Want feature (as shown in Figure 1)；EV71 CPE in RD strengthens with refractivity, becomes round, comes off as principal character (as shown in Figure 2).
And antivirus test respectively organize cell, according to chemicals dilating gradient (dilution one times, totally 12 times), gradient rule every time
Appearance CPE:
A.RSV test group: RSV-MA104 cell before the 5th dilution factor is protected completely, occurs CPE afterwards, and
CPE presents with drug concentration reduction gradually significant rule.
B.EV71 test group: EV71-RD cell before the 9th dilution factor is protected completely, occurs CPE, and CPE afterwards
Present and reduce gradually significantly rule with drug concentration.
By 96 orifice plates of test with 1% neutral red staining, surveying absorbance A by microplate reader at 540nm, each group A value subtracts
After falling virus control group A value, each experimental group A value obtains EC compared with cell controls group A value50, EC50With TC50Compare acquisition TI,
As shown in table 1:
A.RSV test group: suppression RSV occurs CPE, EC at MA104 cell50It is 2-11.2μ g/mL, TI are 955.43.
B.EV71 test group: suppression EV71 occurs CPE, EC at RD cell50It is 2-11.8μ g/mL, TI are 1176.27.
For ease of observing and comparing, foregoing refers to table 1.
Table 1 Hijiki polysaccharide toxin inhibitory test result
As can be seen from the above table, Hijiki polysaccharide crude extract all has significant inhibitory action to RSV, EV71 two-strain,
This prompting Hijiki polysaccharide not only can eliminating phlegm and softening indurated mass, inducing diuresis to remove edema, dispelling heat and resolving dampness, and treatment RSV, EV71 viral infection
There is potential using value in disease areas.The present invention is clinically used for treating disease of viral infection for Hijiki polysaccharide and provides real
Test foundation, the medicine developing anti-RSV and EV71 virus is provided certain directive significance, there is important reference value.
Although the detailed description of the invention of the present invention is described by the above-mentioned accompanying drawing that combines, but not to invention protection domain
Restriction, one of ordinary skill in the art should be understood that, on the basis of technical scheme, those skilled in the art are not required to
Various amendments or deformation that creative work to be paid can be made are the most within the scope of the present invention.
Claims (7)
1. an application for Hijiki polysaccharide, is characterized in that, described Hijiki polysaccharide has antiviral activity, and described Sargassum fusiforme (Harv.) Setch is many
The application of sugar refers to that the Hijiki polysaccharide with antiviral activity resists for the application in RSV or EV71 virus drugs in preparation.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 1, is characterized in that, Hijiki polysaccharide preparation method is that water carries
Alcohol deposition method.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 2, is characterized in that, the concrete steps of described decoction and alcohol sedimentation technique
For, decocting in water after Sargassum fusiforme (Harv.) Setch powder is mixed with distilled water, change liquid and extract, after the filtrate after extracting concentrates, add 5% 3 chloroethene
Acid, centrifugal segregation protein, freezing after the solution dialysis that then will obtain, add 95% ethanol precipitate polysaccharides, centrifuging and taking precipitates,
Vacuum drying i.e. can get Hijiki polysaccharide powder.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 3, is characterized in that, Sargassum fusiforme (Harv.) Setch powder and the quality of distilled water
Ratio is 1:30.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 3, is characterized in that, changing liquid extraction time is 3 times.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 3, is characterized in that, centrifugal speed is 5000r/min, centrifugal
Time is 20-30min.
The application of a kind of Hijiki polysaccharide the most as claimed in claim 3, is characterized in that, dialysis time is 2.5-3.5 days.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610763357.4A CN106309470B (en) | 2016-08-30 | 2016-08-30 | Application of sargassum fusiforme polysaccharide |
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| CN201610763357.4A CN106309470B (en) | 2016-08-30 | 2016-08-30 | Application of sargassum fusiforme polysaccharide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112915099A (en) * | 2018-12-10 | 2021-06-08 | 烟台大学 | Antiviral application of cladophora crude polysaccharide |
| CN113519702A (en) * | 2021-07-17 | 2021-10-22 | 杭州吾尾科技有限公司 | Preparation of carrageen fucoidan and application of carrageen fucoidan in dog and cat foods |
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2016
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112915099A (en) * | 2018-12-10 | 2021-06-08 | 烟台大学 | Antiviral application of cladophora crude polysaccharide |
| CN112915099B (en) * | 2018-12-10 | 2022-02-18 | 烟台大学 | Antiviral application of cladophora crude polysaccharide |
| CN113519702A (en) * | 2021-07-17 | 2021-10-22 | 杭州吾尾科技有限公司 | Preparation of carrageen fucoidan and application of carrageen fucoidan in dog and cat foods |
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| CN106309470B (en) | 2021-05-18 |
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