CN101152207A - Application of laver polyoses in preparing medicament for preventing and treating neurous system disease - Google Patents
Application of laver polyoses in preparing medicament for preventing and treating neurous system disease Download PDFInfo
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- CN101152207A CN101152207A CNA2006100479079A CN200610047907A CN101152207A CN 101152207 A CN101152207 A CN 101152207A CN A2006100479079 A CNA2006100479079 A CN A2006100479079A CN 200610047907 A CN200610047907 A CN 200610047907A CN 101152207 A CN101152207 A CN 101152207A
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Abstract
The invention provides a laver polysaccharide which has the purpose of preventing and treating nervous system diseases. The invention is highly safe and can be taken for a long term.
Description
Technical field
The present invention relates to nerve protection medicine, the application of Porphyra Polysaccharide aspect preparation prevention and treatment nervous system disease medicine specifically.
Background technology
Thallus Porphyrae belongs to Rhodophyta, Bangiales, Bangia fuscopurpurea section Porphyra plant.Known Thallus Porphyrae contains rich in protein, carbohydrate, various vitamin and mineral, have the good reputation of " longevity greens/mustard green " in Japan.The applicating history of Thallus Porphyrae existing more than one thousand years in the traditional Chinese medical science.Record in the Compendium of Material Medica " Thallus Porphyrae cures mainly: the tired plug of steam throat, and liquor is drunk it, and sick goiter beriberi person should eat it.All warts are tied the disease of long-pending piece, and suitable normal food Thallus Porphyrae is an energy hard masses softening and resolving, inducing diuresis to remove edema, the effect that the kidney invigorating is nourished heart ".
Porphyra Polysaccharide is one of main component of Thallus Porphyrae, it is the natural linear polysaccharide of a kind of part of sulfuric acidization of from Thallus Porphyrae, extracting, mainly by galactose, 3, compositions such as 6-inner ether galactose and sulfate, its basic structural unit mainly are the β-D-galactose and 1 that connects by 1,3, the disaccharide unit and 1 that the 4 α-L-galactose that connects-6-sulfates are formed, 3 β-D-galactose that connect and 1, the 4 α-L-3 that connects, the disaccharide unit that 6-inner ether galactose is formed.The C2 of the C6 position of part Porphyra Polysaccharide D-galactose and L-galactose takes place outward partly to methylate in addition.
According to report, Porphyra Polysaccharide has various active, comprises enhancing cellular immunization and humoral immunization, antioxidation, radioprotective, anticancer isoreactivity.Zhang Quanbin etc. have carried out the antioxidation test of Porphyra Polysaccharide to senile rat, prove that it has antioxidation.
Up to the present research, Porphyra Polysaccharide also only are confined to the research and development to the enhancing immunity of general reference, antioxidation, antitumor, the anti-ageing aspect of waiting for a long time, and its research such as mechanism to prevention and treatment nervous system disease aspect does not appear in the newspapers as yet.The drug research of relevant both at home and abroad treatment nervous system disease does not have obvious progress at present, and acquisition extract and effective ingredient become focus and the common recognition of studying in this field from natural plants.
Along with the increase year by year of human average life, the world is just stepping into the global aging epoch.The neurodegenerative disease relevant with the age also increases thereupon, aspect the central nervous system, mainly show as excitement and weaken with process of inhibition, brain function reduces, hypomnesis and forfeiture, the carrying out property that occurs recognition function are subsequently gone down and the increasing the weight of etc. of emotionally disturbed.Alzheimer disease has become the 4th disease after cardiovascular and cerebrovascular disease and the tumor.At present, whole world over-65s crowd person in middle and old age dementia prevalence is about 5%.And alzheimer disease can not make a breakthrough in control in the recent period because of its complicated pathogeny, so the west has the expert to foretell, alzheimer disease will be human first killer of 21 century.
Parkinson disease claim Parkinsonism again, it is the common central nervous system degenerative disease of middle-aged and elderly people, Parkinsonian main pathological characters is because of after the cell generation pathologic change that is arranged in midbrain position " black substance ", the synthetic minimizing of dopamine, the function that suppresses acetylcholine reduces, and then the excitation of acetylcholine strengthens relatively." Parkinsonism " just appearred in the result that both are unbalance.Update shows, in state-owned 1,700,000 Parkinsonians, wherein per 100 people 1 parkinson patient just among the crowd more than 55 years old estimates annual newly-increased about 100,000 people of parkinson patient.
Alzheimer disease and parkinson disease are all brought huge pressure to patient, family and society, make the whole world give numerous concerns to it, the medicine research of neurodegenerative diseases such as senile dementia and parkinson disease becomes an important topic in the geriatrics field.
Summary of the invention
The object of the present invention is to provide the application of Porphyra Polysaccharide aspect preparation prevention and treatment nervous system disease medicine.
For achieving the above object, the technical solution used in the present invention is:
The present invention is carrying out deeply and comprehensively research aspect the anti-aging effects of Porphyra Polysaccharide and the mechanism, find that at last Porphyra Polysaccharide has the protective effect to the neurocyte infringement; Be that Porphyra Polysaccharide can be in the application in preparation prevention and the treatment nervous system disease medicine.
Described nervous system disease comprises parkinson disease or alzheimer disease.
Described Porphyra Polysaccharide is the sulfated polysaccharide that extracts from the marine algae of Rhodophyta Bangiales Bangia fuscopurpurea section Porphyra; Be meant and contain D-galactose and L-galactose-6-sulfate, perhaps also contain 3 simultaneously, the polysaccharide of 6-inner ether-L-galactose.The Porphyra Polysaccharide that the present invention mentioned should be a class polysaccharose substance that obtains by any feasible method.This Porphyra Polysaccharide can be a kind of water extract or other forms of extract; The product that also comprises the different molecular weight that the Porphyra Polysaccharide extract is made through the degraded of any method.
Described Porphyra algae comprises one or more in Porphyra yezoensis Ueda (P.yezoensis), laver (P.tener), porphyra suborbiculata (P.suborbiculata), crisped porphyra algae (P.crispata), long Thallus Porphyrae (P.dentata), porphyra haitanensis (P.haitanensis), the limit Thallus Porphyrae (P.marginata) etc.
The present invention provides a kind of pharmaceutical formulations with neuroprotective activity on the other hand, and said preparation contains the Porphyra Polysaccharide and the pharmaceutically acceptable carrier of above-mentioned any form, as: starch, sodium chloride, microcrystalline Cellulose, sorbic acid and/or mannitol etc.Described preparation comprises the injection type or the local administration preparation of the dosage form of oral administration, non-oral administration.
The present invention has the following advantages:
1. safe, but long expiration is taken.At this stage, the normally irreversible neurodegenerative diseases of alzheimer disease and parkinson disease.In order to keep conditions of patients not continue to worsen, patient's medication cycle is longer.Porphyra Polysaccharide is as a kind of natural marine polysaccharide product that derives from edible seaweed, and its high security can guarantee the needs of patient's long-term prescription.Porphyra Polysaccharide derives from the edible seaweed Thallus Porphyrae, and is very high through the safety of acute toxicity test proof.
2. result of use is good.Experimental result shows that Porphyra Polysaccharide can neuroprotective cell SY5Y antagonism A β
25-35Toxicity has significant neuroprotective; Porphyra Polysaccharide has the activity that improves immunologic function simultaneously, can improve elderly patients' health status from multi-angle.
Description of drawings
Fig. 1 is that Porphyra Polysaccharide is to SY5Y cell A β
25-35The protective effect of damage.**，p＜0.001，*，p＜0.05。
Fig. 2 keeps away the influence of dark test to mice for Porphyra Polysaccharide: Fig. 2 A. is to the influence of errors number, and Fig. 2 B. is to preclinical influence; *, p＜0.05.
Fig. 3 for Porphyra Polysaccharide to the influence of mice water maze test: Fig. 3 A. enters the influence of the errors number of cecum to mice, and Fig. 3 B. finds the preclinical influence of platform to mice; *, p＜0.05.
Fig. 4 is that Porphyra Polysaccharide is to the active influence of acetylcholine transferase (ChAT) in the mice Hippocampus; *, p＜0.05.
Fig. 5 is that Porphyra Polysaccharide is to the active influence of acetylcholinesterase (AChE) in the mice Hippocampus; *, p＜0.05.
The specific embodiment
Below by the specific embodiment the present invention is further specified.Here want the foot pointed out, below the specific embodiment only be used for illustrating the present invention, those skilled in the art are understanding under the prerequisite of spirit of the present invention, can carry out corresponding conversion to the present invention according to the prior art and the knowledge in present technique field, these technical schemes all fall within the scope of the present invention.
The dry Porphyra yezoensis Ueda of 100g is added the water of 40 times of amounts, and 120 ℃ were extracted 3 hours.Silk cover filtering is removed algae-residue, and filtrate is helped filter with kieselguhr.Gained filtrate is concentrated, then this concentrate lyophilizing is obtained the freeze-dry extract of 20g.
Embodiment 2 prepares Porphyra Polysaccharide from porphyra haitanensis
The dry porphyra haitanensis of 100g is added the water of 40 times of amounts, and 120 ℃ were extracted 3 hours.Silk cover filtering is removed algae-residue, and filtrate is helped filter with kieselguhr.Gained filtrate is concentrated, and through tap water flowing water dialysis 2 days, distill water dialysis 1 day then with this dialysis solution 75% ethanol precipitation, with 95% washing with alcohol gained precipitation twice, was dried this precipitation and is obtained 16g oven dry extract.
The preparation of embodiment 3 low-molecular-weight Porphyra Polysaccharide P1
The Porphyra Polysaccharide 5g that gets extraction is made into 1.5% aqueous solution, adds 5mM ascorbic acid and 5mM hydrogen peroxide, normal-temperature reaction 3 hours in this solution.The bag filter that this reactant liquor molecular cut off is 3600Da was dialysed in tap water 2 days, and dialysis is 2 days in distilled water, afterwards vacuum concentration.This concentrate of lyophilizing obtains the Porphyra Polysaccharide sample P 1 that the 3g molecular weight is 35kD.
The preparation of embodiment 4 low-molecular-weight Porphyra Polysaccharide P2
The Porphyra Polysaccharide 5g that gets extraction is made into 1.5% aqueous solution, adds 10mM ascorbic acid and 10mM hydrogen peroxide, normal-temperature reaction 3 hours in this solution.The bag filter that this reactant liquor molecular cut off is 3600Da was dialysed in tap water 2 days, and dialysis is 2 days in distilled water, afterwards vacuum concentration.This concentrate of lyophilizing obtains the Porphyra Polysaccharide sample P 2 that the 3g molecular weight is 6kD.
The preparation of embodiment 5 Porphyra Polysaccharide injections
Get Porphyra Polysaccharide 50g, add water for injection 500ml, mannitol 50g transfers pH value to 7.0, packing, lyophilization.
Embodiment 6 Porphyra Polysaccharide are to the protective effect of neuroblastoma strain SH-SY5Y
The present invention is an index with the cell survival rate, has studied the protective effect of Porphyra Polysaccharide to beta amyloid peptide (β-amyloid peptide, the A β) damage model of neuroblastoma strain SH-SY5Y cell.
Material: the SY5Y cell strain is given by Sweden Caroline SIKA institute Bengt Winblad professor and doctor Pei Jinjing.A β
25-35Segment is synthetic by Xuanwu Hospital of Capital University of Medical Science Beijing brain aging laboratory, the high performance liquid chromatography purification, and purity is more than 98%, serum-free MEM wiring solution-forming, 0.22 μ m considers film sucking filtration ,-20 ℃ of preservations.Porphyra Polysaccharide makes P1 and two samples of P2 that molecular weight is respectively 35KD and 6KD through ascorbic acid and hydrogen peroxide degradation.
Test method: cell culture condition be MEM (Gibco BRL, 41500-067) add in the culture medium 10% hyclone (Hyclone, SH30070.03), 15mmol.L
-1HEPES (DNN company), penicillin 1 * 10
5IU.L
-1, streptomycin 1 * 10
5IU.L
-1, 5%CO
2, 37 ℃, change liquid twice weekly, go down to posterity once.
The MTT metabolic rate is measured: is 1 * 10 with the SY5Y cell with density
3Individual cell inoculation is divided into matched group, A β in 96 orifice plates (Costar product) after 3 days
25-35Group and A β
25-35+ P2 organizes (10,40,80 μ g/ml), A β
25-35+ P1 (10,40,80 μ g/ml) medicine group,, every group 8 hole.Inoculate back 24 hours every holes and add MTT (5mg/ml) 20 μ l, hatched 4 hours for 37 ℃, the sucking-off culture fluid, every hole adds DMSO200 μ l, and jolting 10 minutes is measured 550nm place's optical density value (OD) with microplate reader (Diagnostic Pasteur LP400).
The result: as shown in Figure 1, sample P 1 and P2 all can significantly resist 20 μ M A β when three concentration
25 -35Toxicity improves cultured cell MTT metabolic rate, and strengthens gradually along with sample concentration increases its activity, and the result shows that Porphyra Polysaccharide can neuroprotective cell SY5Y antagonism A β
25-35Toxicity has significant neuroprotective.Neuronal apoptosis plays crucial effect in neural cell injury, medicine can stop it to further develop by intervening apoptotic process, and neurocyte is played a protective role.The SH-SY5Y cell derives from people's neuroblastoma, and its form is similar to normal neurons, tapered with physiological function, and has tangible aixs cylinder, is to study a kind of preferably cell of neurocyte function at present in the world.Porphyra Polysaccharide has neuroprotective to the SY5Y cell, shows that Porphyra Polysaccharide can be used for the treatment of nervous system disease such as parkinson disease and alzheimer disease.
Embodiment 7: the anti-senile dementia effect of Porphyra Polysaccharide
The present invention is an index with acetylcholinesterase and acetylcholine transferase in behavioristics and the brain, and the anti-senile dementia activity of Porphyra Polysaccharide is furtherd investigate.
Laboratory animal: KM mice.Before test, the male KM mice that makes average weight be about 26g freely obtains feedstuff and one week of water to adapt to laboratory environment.
Material: Porphyra Polysaccharide is through ascorbic acid and hydrogen peroxide degradation, and making molecular weight is the P1 sample of 35000Da.
A β
1-40Available from Sigma company, it is dissolved in physiological saline solution, be made into 1mg/ml concentration, sealing is placed in 37 ℃ of incubators and made it cohesion in 7 days.
Test method: after mice adapts to a week, it is weighed and be divided into 5 groups at random, be i.e. three dosage groups of matched group, model group, Porphyra Polysaccharide basic, normal, high (75,150,300mg/kg), positive control drug hydrochloric acid Donna piperazine neat (2.5mg/kg) group.Except that matched group, to the one-sided intracerebral ventricle injection A of mice β
1 -40With reference to the Maurice method, after chloral hydrate anesthesia, ventriculus dexter cerebri location (A:-2mm; L:2mm; H:-4mm), vertical skull inserting needle 3mm slowly injects 3 μ l/ A β only
1-40, matched group is the sterile saline of equivalent.
Animal begins to give relative medicine in second day after operation, dosage 0.5ml/20g, and matched group, model group gavage the equivalent normal saline, and every day 1 time is continuously to the experiment end.
(1) mice is kept away dark test
From A β
1-40Kept away dark test after the modeling on the 9th day.This keep away concealed installation put be divided into bright (25 * 25 * 16cm) dark (and 24 * 20 * 15cm) two compartments, between (8.9 * 11.4cm) link, and door can be shut as required or be opened by the experimenter by an arched door.The compartment floor is rustless steel fence (3.175mm).The fence floor and the stimulator of darkroom one side link, and can shock by electricity, and bright chamber one side does not then link with stimulator, so there is not electric current to pass through.Mice is put into bright chamber, behind the 10s door between the light and shade chamber is opened.After treating that mice (extremity) enters the darkroom fully, immediately middle door is shut, and give once to shock by electricity (electric current is 0.6mA).Allow mice in the darkroom, stop 10s, mice is put back in the cage.The electricity irritation training was put back to bright chamber with mice after 24 hours, and middle door is opened, and mice enters the number of times and the incubation period in darkroom in the record 5min.
(2) rat/mouse labyrinth experiment
From A β
1-40Carried out water maze test, five days by a definite date on the 12nd day after the modeling.This labyrinth has 5 cecums, test first day with a black plastic plate with the labyrinth separated into two parts, make mice only contain the part swimming of a cecum; Tested second day, and reset the position of plastic plate, make mice contain the part swimming of three cecums of row; Test and removed plastic plate in the 3rd day to the 5th day, make the mice covering the race.Write down the errors number of its incubation period of finding platform and arrival cecum respectively.
(3) cortex acetylcholinesterase (AChE) and acetylcholine transferase (ChAT) determination of activity
After behavioristics's EOT, the mice broken end is got brain, peels off hippocampal tissue in ice bath rapidly, makes 10% tissue homogenate with the normal saline of pre-cooling.According to the operation of test kit description, measure the acetylcholinesterase and the acetylcholine transferase activity of this tissue.
The result
Mice is kept away dark test
1) as seen from Figure 2, A β model group is compared mice and is significantly shortened errors number showed increased (p＜0.05) incubation period with three dosage groups of matched group and Porphyra Polysaccharide.
2) rat/mouse labyrinth test
As seen from Figure 3, model group is compared with matched group, and mice finds the incubation period of platform and the errors number of process cecum obviously to prolong (p＜0.05) A β
1-40One-sided intracerebroventricular causes the mice Model of Dementia and sets up successfully.Along with the prolongation of training time, mice incubation period and errors number shorten gradually.Since the 3rd day, A β
1-40One-sided intracerebroventricular model group is compared with three dosage groups of matched group and Porphyra Polysaccharide, the mouse wrong times showed increased, and wherein Porphyra Polysaccharide high concentration group mice is compared remarkable shortening incubation period with model group.Trained the 4th day, the middle and high dosage group of matched group and Porphyra Polysaccharide is compared with model group and is significantly shortened incubation period, and positive control drug hydrochloric acid Donna piperazine produce effects not together shows that the Porphyra Polysaccharide produce effects is fast.Trained the 5th day, three dosage groups of Porphyra Polysaccharide are compared with model group with the neat group of hydrochloric acid Donna piperazine, and the incubation period of mice obviously shortens and errors number obviously reduces.
3) Hippocampus acetylcholinesterase and acetylcholine transferase activity
Acetylcholinesterase and acetylcholine transferase are respectively the catabolic enzyme and the synzyme of acetylcholine, and both regulate the content of acetylcholine in the brain jointly.As shown in Figure 4, in hippocampal tissue, three dosage groups of neat group of hydrochloric acid Donna piperazine and Porphyra Polysaccharide and A β
1-40One-sided intracerebroventricular model group is compared the active rising of acetylcholine transferase, but does not have significance.Matched group, the neat group of hydrochloric acid Donna piperazine are compared with model group with three dosage groups of Porphyra Polysaccharide as shown in Figure 5, acetylcholine esterase active significantly reduces, and what deserves to be explained is that preceding two groups of acetylcholine esterase actives are lower by about 50% than matched group, the Porphyra Polysaccharide of dosage and hydrochloric acid Donna piperazine can both significantly suppress the decomposition of acetylcholine in the Hippocampus together in the explanation, thereby improve the content of acetylcholine in the Hippocampus.This may be that Porphyra Polysaccharide improves A β
1-40One-sided intracerebroventricular causes one of approach of AD symptom.
Conclusion: Porphyra Polysaccharide can A β
1-40Behavioristics's index of the mice of damage is improved the memory of mice, reduces errors number.Improve the acetylcholine transferase activity, reduce acetylcholine esterase active.
Has dementia effect in significant anti-ageing year.
More than having described specific embodiments of the invention, so is not that those skilled in the art can not depart from the improvement and the variation of category of the present invention and spirit to embodiment disclosed herein in order to qualification the present invention.
Claims (7)
1. the application of Porphyra Polysaccharide in preparation prevention and treatment nervous system disease medicine.
2. application according to claim 1 is characterized in that: described nervous system disease comprises parkinson disease or alzheimer disease.
3. application according to claim 1 and 2 is characterized in that: described Porphyra Polysaccharide is the sulfated polysaccharide that extracts from the marine algae of Rhodophyta Bangiales Bangia fuscopurpurea section Porphyra.
4. application according to claim 3 is characterized in that: described Porphyra Polysaccharide is meant and contains D-galactose and L-galactose-6-sulfate, perhaps also contains 3 simultaneously, the polysaccharide of 6-inner ether-L-galactose.
5. application according to claim 3 is characterized in that: described Porphyra algae comprises one or more in Porphyra yezoensis Ueda, laver, porphyra suborbiculata, crisped porphyra algae, long Thallus Porphyrae, porphyra haitanensis, the limit Thallus Porphyrae etc.
6. application according to claim 1 is characterized in that: described medicine is the pharmaceutical composition that contains Porphyra Polysaccharide and pharmaceutically acceptable carrier.
7. application according to claim 6 is characterized in that: the dosage form of described medicine is injection, oral formulations or local administration preparation.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103168784A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院海洋研究所 | Application of porphyra oligosaccharide in disease control of porphyra |
| CN103539863A (en) * | 2012-07-12 | 2014-01-29 | 中国科学院海洋研究所 | Application of low-sulfated heteroglycan prepared from brown alga and rich in glucuronic acid in preparation of medicament and health-care products for treating Parkinson's disease |
| CN106565851A (en) * | 2016-11-22 | 2017-04-19 | 湖州师范学院 | Degradation method of porphyra-haitanensis polysaccharide and anti-senescence activity application thereof |
| CN108210504A (en) * | 2017-12-27 | 2018-06-29 | 中国科学院海洋研究所 | The application of sulphation galactooligosacchari(es and pharmaceutical composition |
| CN111363058A (en) * | 2020-04-08 | 2020-07-03 | 厦门医学院 | Haematococcus polysaccharide and preparation method and application thereof |
| JP2020158476A (en) * | 2019-03-28 | 2020-10-01 | 御木本製薬株式会社 | Amyloid fibril formation inhibitor |
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2006
- 2006-09-27 CN CNA2006100479079A patent/CN101152207A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103168784A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院海洋研究所 | Application of porphyra oligosaccharide in disease control of porphyra |
| CN103168784B (en) * | 2011-12-23 | 2015-03-04 | 中国科学院海洋研究所 | Application of porphyra oligosaccharide in disease control of porphyra |
| CN103539863A (en) * | 2012-07-12 | 2014-01-29 | 中国科学院海洋研究所 | Application of low-sulfated heteroglycan prepared from brown alga and rich in glucuronic acid in preparation of medicament and health-care products for treating Parkinson's disease |
| CN103539863B (en) * | 2012-07-12 | 2016-01-06 | 中国科学院海洋研究所 | The application of the low sulphated heteroglycan being rich in glucuronic acid in preparation treatment anti-parkinson drug and healthcare products in brown alga source |
| CN106565851A (en) * | 2016-11-22 | 2017-04-19 | 湖州师范学院 | Degradation method of porphyra-haitanensis polysaccharide and anti-senescence activity application thereof |
| CN113861306A (en) * | 2016-11-22 | 2021-12-31 | 湖州师范学院 | Degradation method of porphyra haitanensis polysaccharide and anti-aging activity application thereof |
| CN108210504A (en) * | 2017-12-27 | 2018-06-29 | 中国科学院海洋研究所 | The application of sulphation galactooligosacchari(es and pharmaceutical composition |
| JP2020158476A (en) * | 2019-03-28 | 2020-10-01 | 御木本製薬株式会社 | Amyloid fibril formation inhibitor |
| CN111363058A (en) * | 2020-04-08 | 2020-07-03 | 厦门医学院 | Haematococcus polysaccharide and preparation method and application thereof |
| CN111363058B (en) * | 2020-04-08 | 2021-09-28 | 厦门医学院 | Haematococcus polysaccharide and preparation method and application thereof |
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