CN101829140A - Application of astragalus mongholicus polysaccharide as medicine prepared for treating neurodegenerative diseases - Google Patents
Application of astragalus mongholicus polysaccharide as medicine prepared for treating neurodegenerative diseases Download PDFInfo
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Abstract
The invention discloses an application of astragalus mongholicus polysaccharide as the medicine prepared for treating neurodegenerative diseases.Proven by the mammal cell and caenorhadits elegans disease model test, the astragalus polysaccharides has the following functions: effectively inhibiting abnormal aggregation of mammal cell and caenorhadits elegans disease model polyglutamine, effectively relieving neurotoxicity triggered by the abnormal aggregation of polyglutamine and noticeably lengthening life span of the caenorhadits elegans model. With the functions of the astragalus polysaccharides, minor ingredients can be added to the astragalus polysaccharides according to pharmaceutical principles to serve as the medicine for treating neurodegenerative diseases or to be prepared into various dosage forms such as capsule, tablet or granule and the like.
Description
Technical field
The invention belongs to the pharmaceutical technology field.Be particularly related to a kind of application of astragalus polysaccharides in the preparation medicine.
Technical background
Neurodegenerative diseases refers to comprise that Huntingdon disease, Alzheimer disease and Parkinson disease etc. are with specific neurocyte with to organize gradually decline and even death be the PD of principal character.Although also there is not effectively to treat at present the medicine of neurodegenerative diseases, along with the further investigation to this class disease, some micromolecular compounds with potential therapeutical effect are found gradually.Thereby this compounds mainly is by reducing the accumulative level of abnormal protein or disturbing its forming process to prevent from cellular degeneration, change downstream pathologic effect and alleviate its damage that causes to reach the purpose of improving disease symptoms.Wherein, come to gain a special interest in the natural micromolecular compound of plant in sources such as trehalose, resveratrol and curcumin derivate.But at present also not at the bioactive macromolecule of neurodegenerative diseases as medicinal application in clinical.
The Radix Astragali is the traditional tonification class Chinese medicine of China, and medicinal Astragalis is the dry root of the perennial leguminous herbaceous plant film folder Radix Astragali and Radix Astagali.The main effective ingredient of the Radix Astragali is polysaccharide, saponins and flavonoid.Found that at present astragalus polysaccharides has antitumor, defying age, antiviral, wide biological activity such as treatment diabetes and cardiovascular disease, but the Radix Astragali is also very not deep in the research in neurodegenerative diseases field.There are some researches show; the hippocampal neurons injury that Radix Astragali extract can protect A β to be caused; astragaloside has protective effect to the mouse brain damage that A β is caused; astragalus polysaccharides can reduce the toxicity of the radical pair neurogliocyte that the levodopa oxidation produces, thereby and the expression that can regulate and control rat IGF-1 protect cerebellum and nucleus basalis neuron with secretion.But whether astragalus polysaccharides can effectively suppress the poly glumine gathering and alleviate poly glumine to assemble toxicity, yet there are no report.
Summary of the invention
Order of the present invention is exactly the developing direction of astragalus polysaccharides being utilized at above-mentioned prior art, excavates the function of astragalus polysaccharides aspect the treatment neurodegenerative diseases, and utilizes its preparation treatment neurodegenerative diseases medicine.
Technical scheme of the present invention is: utilize astragalus polysaccharides can suppress the unusual gathering of poly glumine and effectively alleviate to accumulate in the neurovirulent function of bringing out in the neurodegenerative diseases unusually by poly glumine, the neurodegenerative diseases medicine is treated in preparation according to the pharmacy principle.
Above-mentioned treatment neurodegenerative diseases medicine comprise treatment Huntingdon disease, Alzheimer disease and Parkinson disease etc. with specific neurocyte and to organize decline gradually and even death be the PD medicine of principal character.
The present invention shows: astragalus polysaccharides has the unusual gathering of poly glumine in effective inhibition mammalian cell and the beautiful nematicide body; Neurotoxicity that brings out by the unusual gathering of poly glumine in the beautiful nematicide body of effective alleviation and the life-span of the beautiful nematicide of significant prolongation.Utilize above-mentioned functions, in astragalus polysaccharides, add the medicine that can be used as the treatment neurodegenerative diseases behind the adjuvant according to the pharmacy principle, and can be made into various dosage forms such as capsule, tablet or granule.
Description of drawings
Accompanying drawing 1 suppresses the unusual accumulative data characterization figure of poly glumine in the COS-7 cell for astragalus polysaccharides.Characterize 0.5mg/ml respectively, 2mg/ml and 5mg/ml astragalus polysaccharides reduce the protein of COS-7 cell HDQ74 mediation and assemble situation (* p＜0.05 unusually; * p＜0.005).X is astragalus polysaccharides concentration (mg/ml), and A is the ratio (%) that the COS-7 cell that contains the poly glumine accumulation point accounts for total cell number.
Accompanying drawing 2 suppresses the unusual accumulative data characterization figure of poly glumine in the beautiful nematicide body of AM141 for astragalus polysaccharides.Characterize the paraprotein gathering situation (* * * p＜0.0001) that the 5mg/ml astragalus polysaccharides significantly reduces the beautiful nematicide body wall of the AM141 Musclar layer that is mediated by polyQ40.X is astragalus polysaccharides concentration (mg/ml), and B contains the ratio (%) that accounts for beautiful line insect number greater than the beautiful nematicide of 10 poly glumine accumulation points.
Accompanying drawing 3 astragalus polysaccharidess significantly reduce the beautiful nematicide ASH of HA759 neuronal death.Characterize 1mg/ml respectively, the 2.5mg/ml astragalus polysaccharides significantly reduces the situation (* p＜0.05) by the beautiful nematicide ASH of the HA759 neuronal death of HDQ150 mediation.X is astragalus polysaccharides concentration (mg/ml), and C is the ratio (%) that the beautiful nematicide of ASH neuronal survival accounts for beautiful line insect number.
The specific embodiment
Unusual gathering, the astragalus polysaccharides that unusual gathering, the astragalus polysaccharides that astragalus polysaccharides is suppressed poly glumine in the mammalian cell below in conjunction with the specific embodiment suppresses poly glumine in the beautiful nematicide body alleviated in the beautiful nematicide body by the unusual neurotoxicity that brings out and the astragalus polysaccharides assembled of poly glumine the situation that influences in beautiful nematicide life-span is described further:
One, the preparation of astragalus polysaccharides
1. Milkvetch Root (purchasing the Wu Handian in Beijing Tongrentang) is pulverized the back and is crossed the 0.85mm sieve;
2. take by weighing radix astragali coarse powder 100g;
3.500ml petroleum ether, 70 ℃, reflux, extract, 1h 2 times, volatilizes medical material behind the sucking filtration;
4.500ml 95% ethanol, 80 ℃, reflux, extract, 2h 2 times, volatilizes medical material behind the sucking filtration;
5.800ml water, 95 ℃, extract 2h, 1 time, sucking filtration is collected extracting solution;
6.600ml water, 95 ℃, extract 1.5h, 2 times, sucking filtration is collected extracting solution, merges 3 times extracting solution;
7. centrifugal: 5000rpm, 15min, room temperature; Abandon precipitation, get the supernatant extracting solution;
8. sucking filtration supernatant extracting solution revolves steaming and is concentrated in the 100ml;
9. 95% ethanol that extracting solution is dropwise added magnetic agitation makes that the ethanol final concentration is 80%;
10. collecting precipitation is dissolved in 100ml one-level water, repeating step 9-10 step 2 time;
11. it is centrifugal: 5000rpm, 15min, room temperature; Collecting precipitation is dissolved in 100ml one-level water;
12. according to polysaccharide solution: Sevage reagent [chloroform: n-butyl alcohol=5: 1 (V: V)]=5: 1 (V: V) add Sevage reagent, shake well 20min;
13. it is centrifugal: 5000rpm, 5min, room temperature; The careful polysaccharide supernatant of collecting;
14. repeating step 12-13 step 2 time;
15. supernatant is dropwise added the dehydrated alcohol of magnetic agitation, makes that the ethanol final concentration is 80%;
16. it is centrifugal: 5000rpm, 15min, room temperature; Collecting precipitation with the repeatedly washing on a small quantity of dehydrated alcohol, acetone and ether, dissolves with suitable quantity of water successively;
17. obtain astragalus polysaccharides after the lyophilization, yield 3.32%.
Two, astragalus polysaccharides suppresses the unusual gathering of poly glumine in the mammalian cell
1.COS-7 cell culture and EGFP-HDQ74 transfection
(1) with containing 10% hyclone, 100U/ml penicillin and streptomycin, and the DMEM culture medium of 2mM L-glutaminate, at 37 ℃ and 5%CO
2Cultivate the COS-7 cell under the condition;
When (2) cell grows into the 80%-90% degrees of fusion, hatch the 8-10min peptic cell for 37 ℃ with 1.5ml 2 * trypsin solution;
(3) with 3-5 * 10
4Density with cell inoculation to every hole be placed with coverslip and contain 2ml DMEM culture medium 6 orifice plates in, at 37 ℃ and 5%CO
2Cultivate 24h under the condition;
(4) when cell grows to the 60%-80% fusion on coverslip, culture medium is abandoned in suction, add 900 μ l fresh cultures, and add the DMEM culture medium that the 100 μ l that contain 0.75 μ g EGFP-HDQ74 plasmid DNA and 4.5 μ l transfection reagents do not contain serum, at 37 ℃ and 5%CO
2Cultivate 3h under the condition;
(5) add the 1ml fresh culture, at 37 ℃ and 5%CO
2Cultivate 48h under the condition;
(6) inhale and to abandon culture medium,, add 0.75ml 4% paraformaldehyde, at room temperature fixing 30min with 1ml 1X PBS flushing 1 time;
(7) inhale and to abandon the paraformaldehyde fixative,, draw 20 μ l DAPI in microscope slide central authorities with 1ml 1X PBS flushing 3 times, with the coverslip mounting in microscope slide central authorities;
(8) the COS-7 cell after the mounting at room temperature can be under fluorescence microscope be observed the EGFP positive cell and is assembled the formation situation by green fluorescence, thereby and judges the quantity of living cells by the form of blue-fluorescence observation of cell nuclear.EGFP positive cell kytoplasm manifests uniform green fluorescence, putting before this when occurring very bright green point-like thing in the born of the same parents, then for having formed the abnormal protein aggregating cells.At random at least 200 EGFP positive cells are observed, statistics contains assembles the number of cells that forms.If a cell does not contain accumulation point, count 0; If a cell contains one or more accumulation point, then count 1.Assemble cell percentage=gathering number of cells/EGFP positive cell number * 100%.Nucleus after the DAPI dyeing is ellipticity, manifests uniform blue-fluorescence, if nucleus manifests fragmentation or pyknosis to less than apoptosis forms such as 50% normal nuclear diameter by blue-fluorescence, then thinks cell death, with this canonical statistics cell mortality.Total cellular score * 100% of cell mortality=cell death number/random statistical.
(9) gathering cell percentage and cell mortality after the HDQ74-EGFP transfection are added up.
2. astragalus polysaccharides acts on the COS-7 cell after EGFP-HDQ74 mediates, and suppresses intracellular abnormal protein and assembles
(1) 50mg/ml astragalus polysaccharides mother solution is dissolved in the 1ml culture medium of step 1 (5) according to certain volume, is mixed with final concentration and is respectively 0.5mg/ml, the culture medium that contains astragalus polysaccharides of 2mg/ml and 5mg/ml is according to above-mentioned 1 (3)-(7) step process cell;
(2) behind the culture medium processing cell with the variable concentrations astragalus polysaccharides, the gathering cell percentage that is mediated by EGFP-HDQ74 in the COS-7 cell obtains reduction in various degree, and the astragalus polysaccharides concentration of 2mg/ml suppresses best (* p＜0.05 of accumulative effect; * p＜0.005) (Fig. 2).
Three, astragalus polysaccharides suppresses the unusual gathering of poly glumine in the beautiful nematicide body
1. the cultivation of beautiful nematicide
(1) beautiful nematicide N2 of wild type and the beautiful nematicide AM141 of transgenic and HA759 all purchase in Univ Minnesota-Twin Cities USA beautiful nematicide hereditism center;
(2) be distributed on the NGM plate as applying foodstuffs with escherichia coli OP50, cultivate beautiful nematicide at 15 ℃-25 ℃ according to the experiment needs.
2. beautiful nematicide synchronization growth
(1) as stated above the beautiful nematicide of NGM plate culture medium, obtains a large amount of egg-laying season adults;
(2) with M9 solution beautiful nematicide is collected in the 1.5ml centrifuge tube, 2000rpm, 2min, room temperature is centrifugal, and M9 solution washing 3 times is to remove OP50;
(3) usefulness M9 solution at 0.5ml, adds 1ml Bleaching solution with worm liquid standardize solution, and jolting 3-5min in the microscopically monitoring, a large amount of worm's ovums occurs in the complete cracking of beautiful nematicide, solution up and down;
(4) 4000pm, 2min, room temperature is centrifugal, discards Bleaching solution as far as possible, and with M9 solution washing worm's ovum 3 times;
(5) worm's ovum is transferred in the 50ml conical flask,, cultivates the L1 phase larva after 12-20h can obtain synchronization in 20 ℃ with 3ml S solution 120rpm.
3.AM141 the detection method of beautiful nematicide model: focusing experiment
(1) preparation agarose cushion;
(2) the 20mM Hydrazoic acid,sodium salt is fixed beautiful nematicide;
(3) observation of beautiful nematicide: the polypide that is fixed on the agarose cushion is observed one by one with Olympus IX51S8F fluorescence inverted microscope, add up the quantity of the fluorescence accumulation point on each polypide somatic cell parietal cell, at random at least 100 insects are added up, with 10 accumulation points is threshold value, add up respectively greater than the individual amount of 10 accumulation points with less than the individual amount of 10 accumulation points, greater than the beautiful nematicide percentage rate of 10 accumulation points=greater than the individual amount of 10 accumulation points/(greater than the individual amount of 10 accumulation points+) * 100% less than the individual amount of 10 accumulation points.
4. the body wall Musclar layer protein of poly glumine mediation is assembled unusually in the beautiful nematicide of astragalus polysaccharides inhibition AM141
(1) with the density of the beautiful nematicide of L1 phase AM141 behind the S solution adjustment synchronization, 10 μ l only are controlled at 10-15;
(2) the OP50 suspension of adding centrifugation is an amount of, makes that the OD value of worm liquid is 0.6;
(3) get 96 orifice plates of cleaning sterile, 50mg/ml astragalus polysaccharides mother solution and contrast S solution are joined the sample hole with 10 μ l/ holes, multiple 3 holes of each sample counterpoise, in each hole, add the worm liquid (astragalus polysaccharides sample hole final concentration is 5mg/ml) that 90 μ l have adjusted insect density and OP50OD value again, 96 orifice plates are added a cover and are sealed with Parafilm, put 20 ℃ of incubators and cultivate 48h;
(4) situation of change of accumulation point in the AM141 body wall muscle cell after the statistics astragalus polysaccharides is handled.The result shows that the protein that can obviously suppress behind the 5mg/ml astragalus polysaccharides effect 48h by the beautiful nematicide AM141 transgenic strain body wall Musclar layer of YFP-polyQ40 mediation assembles (* * * p＜0.0001) (Fig. 3) unusually.
Four, astragalus polysaccharides is alleviated in the beautiful nematicide body by the unusual neurotoxicity that brings out of assembling of poly glumine
1.HA759 the cultivation of the beautiful nematicide strain of transgenic and synchronization growth are with the step 1 and 2 of embodiment 3.
2.HA759 the detection method of beautiful nematicide model: ASH neuron survival rate experiment
(1) fixing means of the preparation of agarose cushion and beautiful nematicide is with the step 4 among the embodiment 3 (1) and (2).
(2) observation of beautiful nematicide: with Olympus IX51S8F fluorescence inverted microscope the polypide that is fixed on the agarose cushion is observed one by one, the visual field is focused on the nerve ring position of pharyngeal rear side.Select the by-level side of veutro and dorsal part to observe nematicide, can be observed the ASH and the ASI neuron of homonymy, the ASH neuron is positioned at the neuronic downside of ASI.Along with the growth of beautiful nematicide, the influence that the ASH neuron is subjected to the neurotoxicity of GFP-polyQ150 mediation causes death, thereby the green fluorescence of its expression disappears, and the ASI neuron is not influenced by it, still can be observed green fluorescence.At random at least 100 beautiful nematicides are observed, add up ASH neuronal survival and dead nematode population respectively, the beautiful nematode population of the beautiful nematicide percentage rate=ASH neuronal survival of ASH neuronal survival/(the beautiful nematode population of the beautiful nematode population+ASH neuronal death of ASH neuronal survival) * 100%.
3. astragalus polysaccharides suppresses the neurotoxicity of the beautiful nematicide of HA759 by the poly glumine mediation
(1) with the density of L1 phase HA759 nematicide behind the S solution adjustment synchronization, 10 μ l only are controlled at 10-15;
(2) the OP50 suspension of adding centrifugation is an amount of, makes that the OD value of worm liquid is 0.6;
(3) 50mg/ml astragalus polysaccharides mother solution is diluted successively be 25mg/ml, 10mg/ml and 2.5mg/ml sample;
(4) get 96 orifice plates of cleaning sterile, the astragalus polysaccharides sample and the contrast S solution of three kinds of concentration are joined the sample hole with 10 μ l/ holes, multiple 3 holes of each sample counterpoise, adding 90 μ l again in each hole has adjusted the worm liquid of insect density and OP50OD value (astragalus polysaccharides sample hole final concentration has been followed successively by 2.5mg/ml, 1mg/ml and 0.25mg/ml), 96 orifice plates are added a cover and are sealed with Parafilm, put 15 ℃ of incubators and cultivate 72h;
(5) situation of the HA759 nematicide ASH neuronal survival after the effect of statistics astragalus polysaccharides, the result can obviously slow down the beautiful nematicide neurotoxicity that is mediated by GFP-HDQ150 after showing 2.5mg/ml and 1mg/ml astragalus polysaccharides effect 72h, and the ASH neuronal survival obviously raises (* p＜0.05) (Fig. 4).
Five, astragalus polysaccharides is to the influence in beautiful nematicide life-span
1. life experiment
(1) adjust the density of L1 each nematicide of phase behind the synchronization with S solution, 10 μ l only are controlled at 10-30, prevent since worm density too high cause growing inhomogeneous;
(2) the OP50 suspension of adding centrifugation is an amount of, makes that the OD value of worm liquid is 0.6;
(3) adjust worm density with S solution behind 120rpm and 20 ℃ of shaking tables cultivation 42-45h, 90 μ l only are controlled at 10-15; The OP50 suspension that adds centrifugation is an amount of, makes that the OD value of worm liquid is 0.8; According to worm liquid total amount add proper volume 50mM 5-FuDR (2 '-Deoxy-5-Fluorouridine), 50mg/ml Carbenicillin and 0.25mg/ml amphotericin mother solution, make its final concentration be respectively 0.5mM, 50 μ g/ml and 0.1 μ g/ml;
(4) cultivate 24h at 120rpm and 20 ℃ of shaking tables;
(5) get 96 orifice plates of cleaning sterile, each sample and contrast S solution are joined the sample hole with 10 μ l/ holes, multiple 5 holes of each sample counterpoise, adding 90 μ l have adjusted the worm liquid of beautiful nematode density and OP50OD value in each hole again; Remain the hole behind the end of the sample on 96 orifice plates and fill up, add a cover and seal, put 20 ℃ of incubators and cultivate with Parafi lm with 100 μ l sterilized water;
(6) above sample was to calculate in 0 day the same day, added up the borer population amount alive in each hole of sample since the 1st day, statistics once every other day, finish to add up to all beautiful nematicides are all dead, the natural law (average life of beautiful nematicide) of correspondence is as the index of life-span variation before and after the statistics administration during with 50% individual survival.
2. astragalus polysaccharides prolongs the life-span of N2, AM141 and the beautiful nematicide of HA759
(1) 50mg/ml astragalus polysaccharides mother solution is diluted to 25mg/ml successively, 10mg/ml and 2.5mg/ml gradient concentration sample;
(2) according to the method described above N2, AM141 and the beautiful nematicide of HA759 are carried out life experiment, the astragalus polysaccharides final concentration is respectively 2.5mg/ml, 1mg/ml and 0.25mg/ml;
(3) result shows, the variable concentrations astragalus polysaccharides all has the effect of life-saving to above-mentioned three kinds of beautiful nematicides, and wherein the effect of lengthening the life of 1mg/ml astragalus polysaccharides is the most obvious, life-spans of above-mentioned three kinds of beautiful nematicides is prolonged reach 29.6%, 50% and 64.2% respectively.
Claims (2)
1. astragalus polysaccharides is as the application of preparation treatment neurodegenerative diseases medicine, it is characterized in that: utilize astragalus polysaccharides can suppress the unusual gathering of poly glumine and effectively alleviate to accumulate in the neurovirulent function of bringing out in the neurodegenerative diseases unusually by poly glumine, the neurodegenerative diseases medicine is treated in preparation according to the pharmacy principle.
2. astragalus polysaccharides according to claim 1 is characterized in that as the application of preparation treatment neurodegenerative diseases medicine: described treatment neurodegenerative diseases medicine comprise treatment Huntingdon disease, Alzheimer disease and Parkinson disease etc. with specific neurocyte and to organize decline gradually and even death be the PD medicine of principal character.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103127286A (en) * | 2011-12-05 | 2013-06-05 | 鲍力恒 | Medicine component used for curing or lightening melancholia or neurodegeneration disease |
| CN103525858A (en) * | 2013-09-29 | 2014-01-22 | 深圳大学 | Establishment method of plant cell model of high throughput screening anti-Huntington's disease active substance and application of plant cell model |
| CN103564288A (en) * | 2013-10-12 | 2014-02-12 | 无限极（中国）有限公司 | Application of madder polysaccharide in preparation of health-protection food with effects of preventing and treating neurodegenerative diseases |
| CN103751204A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for preventing vascular dementia and application thereof |
| CN103751206A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for preventing Alzheimer's disease and application thereof |
| CN103751205A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for treating vascular dementia and application thereof |
| CN106614426A (en) * | 2016-12-06 | 2017-05-10 | 中山大学 | Vitro culture medium and vitro culture method of Angiostrongyluscantonensis |
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| CN103127286A (en) * | 2011-12-05 | 2013-06-05 | 鲍力恒 | Medicine component used for curing or lightening melancholia or neurodegeneration disease |
| CN103525858A (en) * | 2013-09-29 | 2014-01-22 | 深圳大学 | Establishment method of plant cell model of high throughput screening anti-Huntington's disease active substance and application of plant cell model |
| CN103525858B (en) * | 2013-09-29 | 2015-12-09 | 深圳大学 | The establishment method of the vegetable cell model of high flux screening anti-huntington disease activity material and application thereof |
| CN103564288A (en) * | 2013-10-12 | 2014-02-12 | 无限极（中国）有限公司 | Application of madder polysaccharide in preparation of health-protection food with effects of preventing and treating neurodegenerative diseases |
| CN103564288B (en) * | 2013-10-12 | 2015-10-28 | 无限极（中国）有限公司 | Madder polysaccharide has the application in the health food of control nerve degenerative diseases effect in preparation |
| CN103751205B (en) * | 2014-01-21 | 2015-09-23 | 新乡医学院 | The pharmaceutical composition for the treatment of vascular dementia and application thereof |
| CN103751205A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for treating vascular dementia and application thereof |
| CN103751206A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for preventing Alzheimer's disease and application thereof |
| CN103751204A (en) * | 2014-01-21 | 2014-04-30 | 新乡医学院 | Medicinal composition for preventing vascular dementia and application thereof |
| CN103751204B (en) * | 2014-01-21 | 2016-01-20 | 新乡医学院 | The pharmaceutical composition of prevention vascular dementia and application thereof |
| CN106614426A (en) * | 2016-12-06 | 2017-05-10 | 中山大学 | Vitro culture medium and vitro culture method of Angiostrongyluscantonensis |
| CN106614426B (en) * | 2016-12-06 | 2019-10-29 | 中山大学 | A kind of angiostrongylus cantonensis in-vitro culture medium and extracorporeal culturing method |
| CN112353812A (en) * | 2020-12-14 | 2021-02-12 | 湖州师范学院 | Application of Gracilaria lemaneiformis polysaccharide in treating and preventing Huntington disease |
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