CN103609937A - Application of acanthopanax senticosus polysaccharide in preparation of neuroprotective health food - Google Patents
Application of acanthopanax senticosus polysaccharide in preparation of neuroprotective health food Download PDFInfo
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- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
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- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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Abstract
The invention discloses an application of acanthopanax senticosus polysaccharide in preparation of a neuroprotective health food. The acanthopanax senticosus polysaccharide is obtained by degreasing, water extraction, alcohol precipitation, ion exchange and dialysis processes, and is characterized in that the preparation method is simple, stable and efficient, and industrial production can be carried out conveniently. According to the mammalian cell and caenorhabditis elegans disease model test, acanthopanax senticosus polysaccharide is able to effectively inhibit cytotoxicity of neurotoxicity compounds and relieve neurotoxicity caused by abnormal gathering of polyglutamine in a caenorhabditis elegans disease model, also obviously lowers the level of in-vivo reactive oxide species in the caenorhabditis elegans disease model, increases the survival rate of caenorhabditis elegans under oxidative stress, and shows a good neuroprotective effect.
Description
Technical field
The invention belongs to healthy food/health food technology field, be specifically related to Radix Et Caulis Acanthopanacis Senticosi polysaccharide and there is the application in the health food of neuroprotective effect in preparation.
Technical background
Aging is a kind of inevitable natural phenomena, easily causes the various diseases such as angiocardiopathy, cancer, the nervous system disease.Wherein, the nervous system disease pathogenic factor is that carrying out property of neuron sex change death causes nervous system impaired, major effect patient's cognition and motor function, and the fatal rate of disabling is high.Along with the aggravation of population in the world aging degree, great cerebral disease ，Gei patient family and society that the nervous system disease has become serious harm people life and health and quality of life bring great psychology and financial burden.Although the exploitation of this type of disease therapeuticing medicine is dropped into day by day and is increased; but there is no at present effective medicine; and it is occurred in earlier stage to evolution lack to be paid close attention to and prevention, so healthy food/health food that active development has a neuroprotective effect will have great social effect to preventing and treating the nervous system disease.
Quantity research shows greatly, and the generation development of multiple the nervous system disease all relates to the abnormal gathering of protein and assembles the Stress Factors such as toxicity, and for example, the main pathology mark sheet of Huntington's chorea is now by poly glumine (polyglutamine in cell; PolyQ) the abnormal Huntington protein inclusion body causing that extends.These protein are assembled the product of formation extremely by various abiotic stress reactions such as induced oxidation, inflammation, cause neure damage and produce nervous function decline.Therefore, by suppressing the abnormal gathering of disease protein matter and alleviate the toxicity such as oxidation that caused by different polyprotein to coerce damage by the Critical policies that is control neurological conditions.
China's Traditional Chinese Medicine is accumulating abundant research and clinical experience aspect old and feeble and diseases associated with senescence, and multiple Chinese herbal medicine is for a long time for delaying senility and the purposes such as neuroprotective.For example, wilsonii is regarded as having the good medicine that adds essence benefit marrow and anti-aging effects from ancient times.The Li Shizhen (1518-1593 A.D.) < < of Ming Dynasty Compendium of Materia Medica > > claims that " good with the five leaf persons of accompanying each other, therefore named slender acanthopanax, has another name called five flowers to wilsonii.Slender acanthopanax is controlled rheumatism, strengthens the bone, and its merit is good dark, and Ning get Yi is slender acanthopanax, need not the full car of gold and jade ", with the saying of " wine made in article, can become its taste, with Jin Maicao, do not say that it is expensive ", wilsonii has been done to very high praise again.Modern study shows, Radix Et Caulis Acanthopanacis Senticosi polysaccharide is one of main pharmacodynamics composition of wilsonii, has anti-oxidant, radioresistance, immunological regulation and the various biological such as antitumor is active, in healthcare field, has huge development and application potentiality.Yet these researchs concentrate on antitumor and anti-oxidant aspect mostly, the research to Radix Et Caulis Acanthopanacis Senticosi polysaccharide in neuroprotective and the nervous system disease field and application are paid close attention to not enough.Therefore, strengthen Radix Et Caulis Acanthopanacis Senticosi polysaccharide and in the research aspect neuroprotective effect, be not only conducive to widen the range of application of itself, new active material can also be provided for the control of the nervous system disease, and can promote polysaccharide in the deep application in medicines and health protection field.
Summary of the invention
The object of the present invention is to provide Radix Et Caulis Acanthopanacis Senticosi polysaccharide to there is the application in the health food (health food is also referred to as healthy food or functional food) of neuroprotective effect in preparation.
Wherein said neuroprotective effect refer to take protein gathering extremely, oxidative stress and the decline of specific nerve cell dead be the preventive and therapeutic effect of the nervous system disease of main pathological characters.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide described in the present invention preferably prepares by the following method: take wilsonii as raw material; through preparing Radix Et Caulis Acanthopanacis Senticosi polysaccharide containing operations such as degreasing, water extraction, alcohol precipitation, ion-exchange and dialysis; the Radix Et Caulis Acanthopanacis Senticosi polysaccharide preparing has neuroprotection, can prepare the health food with neuroprotective effect.
The present invention is in the preparation process of Radix Et Caulis Acanthopanacis Senticosi polysaccharide, and the design parameter of each operation is as follows:
During degreasing, employing ethanol is degreasing solvent, and the mass volume ratio of wilsonii and ethanol is 1:2g/mL～1:20g/mL, and skimming temp is 50～90 ℃, and the time is 1～10h; During water extraction, the mass volume ratio of wilsonii and water is 1:2g/mL～1:20g/mL, and extracting temperature is 50～100 ℃, and water extraction time is 1～10h, after water extraction is got, obtains extract; During alcohol precipitation, adopt ethanol precipitation extract, the volume ratio of ethanol and extract is 3:1～5:1.
Ion-exchange is anion exchange, after alcohol hypostasis is dissolved, crosses anion-exchange column, adopts successively water and eluent to carry out wash-out, and eluent is crossed bag filter dialysis, and dislysate, through concentrated and dry processing, obtains Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
The material of described anion exchange is preferably cellulose, sephadex, Ago-Gel or resin; The molecular cut off of bag filter is 1.0～5.0kD.
During anion exchange, eluent is preferably acetate, phosphate, barbiturate, citrate, sodium salt, sylvite, ammonium salt or glycine.
Wilsonii described in the present invention preferably refers to medicinal part root or the root skin of wilsonii.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide provided by the invention has neuroprotective effect and refers to that it can significantly suppress the cytotoxicity of neurotoxicity compound in cell and whole animal level, suppress the abnormal neurotoxicity causing of assembling of poly glumine in C. Elegans Automatic Screening disease model, can also significantly reduce C. Elegans Automatic Screening disease model activity in vivo oxygen free radicals and extend its survival rate under oxidative stress, thereby can be used for control and comprise extremely assembling with protein of Alzheimer's disease and Huntington's chorea etc., oxidative stress and specific nerve cell are the nervous system disease of main pathological characters with organizing decline death gradually.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide purposes provided by the invention is also included in preparation and has the healthy food of neuroprotective effect or the application in health food.Above-mentioned Radix Et Caulis Acanthopanacis Senticosi polysaccharide is added to other raw material if lactalbumin, auxiliary material are as maltodextrin, can be prepared as healthy food or the health food with neuroprotective effect.
Tool of the present invention has the following advantages:
(1) the Radix Et Caulis Acanthopanacis Senticosi polysaccharide preparation technology that prepared by the present invention is simply perfect, and stability and high efficiency is applicable to suitability for industrialized production;
(2) the Radix Et Caulis Acanthopanacis Senticosi polysaccharide that prepared by the present invention verified its mammalian neural cell with in C. Elegans Automatic Screening whole animal level, the abnormal gathering of the relevant protein of the nervous system disease and neurotoxicity thereof, oxidative stress damage are all had to inhibitory action; show that it can be applicable to have in the solid-liquid preparation healthy food or health food of neuroprotective effect; for control and the successful aging of the nervous system disease provides New Scheme, and can promote deep application and the modernization of Chinese medicine development of polyose.
Accompanying drawing explanation
Below by the specific embodiment and pharmacodynamic study result, the invention will be further described.
Fig. 1 is that in embodiment 4, Radix Et Caulis Acanthopanacis Senticosi polysaccharide suppresses respectively A β 42, 3-nitropropionic acid, the Cytotoxic data characterization figure of Pidolidone and paraquat, wherein Figure 1A characterizes 1mg/mL, 2mg/mL and 4mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide are alleviated the cytotoxicity of 42 couples of HNB's cell SH-SY5Y of A β, Figure 1B characterizes 1mg/mL, 2mg/mL and 4mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide are alleviated the cytotoxicity of 3-nitropropionic acid to SH-SY5Y, Fig. 1 C characterizes 1mg/mL, 2mg/mL and 4mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide are alleviated the cytotoxicity of Pidolidone to SH-SY5Y, Fig. 1 D characterizes 1mg/mL, 2mg/mL and 4mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide are alleviated the cytotoxicity of paraquat to SH-SY5Y, wherein * represents p<0.05,
Fig. 2 is that in embodiment 5, Radix Et Caulis Acanthopanacis Senticosi polysaccharide suppresses poly glumine abnormal data characterization and micro-schematic diagram of assembling toxicity in C. Elegans Automatic Screening body, wherein Fig. 2 A characterizes the neuronic existing state of HA759 C. Elegans Automatic Screening ASH unprocessed and that process through 2mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide, Fig. 2 B characterizes 2mg/mL and 4mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide suppresses poly glumine gathering to the neuronic neurotoxicity of HA759 C. Elegans Automatic Screening ASH, Fig. 2 C characterizes 2mg/mL Radix Et Caulis Acanthopanacis Senticosi polysaccharide and improves the ASH neuron behavioral function that poly glumine is assembled toxic damages, wherein * represents p<0.05,
Fig. 3 is the data characterization figure that in embodiment 6, Radix Et Caulis Acanthopanacis Senticosi polysaccharide reduces this model oxidative stress survival rate of the gentle raising of activated oxidized water in C. Elegans Automatic Screening poly glumine model, wherein Fig. 3 A characterizes 2mg/ml Radix Et Caulis Acanthopanacis Senticosi polysaccharide reduction HA759 C. Elegans Automatic Screening activity in vivo oxygen free radicals, and Fig. 3 B characterizes 2mg/ml Radix Et Caulis Acanthopanacis Senticosi polysaccharide and improves the survival rate of HA759 C. Elegans Automatic Screening under paraquat oxidative stress.
The specific embodiment
The present invention will be described to enumerate a part of specific embodiment below, is necessary to be pointed out that at this following specific embodiment, only for the invention will be further described, does not represent limiting the scope of the invention.Some nonessential modifications that other people make according to the present invention and adjustment still belong to protection scope of the present invention.
A kind of Radix Et Caulis Acanthopanacis Senticosi polysaccharide that the present embodiment provides, it prepares by the following method: after Radix Acanthopanacis Senticosi root and root skin are pulverized according to solid-liquid ratio 1:5(g/mL) add ethanol, heat stirring and refluxing 3h at 60 ℃, filtration under diminished pressure, except desolventizing, reclaims medicinal material.According to solid-liquid ratio 1:15(g/mL) add water, at 90 ℃, add thermal agitation and extract 8h.After reduced pressure concentration, centrifugal acquisition clarification extract, adds 4 times of volume ethanol in extract according to 1:4 volume ratio, spends the night and staticly settles, centrifugal collecting precipitation process vacuum freeze drying after mixing at 4 ℃.By the water-soluble rear DEAE-sepharose sephadex anion-exchange column of crossing of sediment, adopt successively water and 1.0mol/L sodium chloride solution wash-out, collect the eluent of 1.0mol/L sodium chloride solution, adopt 1.0kD bag filter to dialyse, dialysis is dammed, and liquid passes through reduced pressure concentration and vacuum freeze drying obtains Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
A kind of Radix Et Caulis Acanthopanacis Senticosi polysaccharide that the present embodiment provides, it prepares by the following method: after Radix Acanthopanacis Senticosi root and root skin are pulverized according to solid-liquid ratio 1:10(g/mL) add ethanol, heat stirring and refluxing 5h at 70 ℃, filtration under diminished pressure, except desolventizing, reclaims medicinal material.According to solid-liquid ratio 1:5(g/mL) add water, at 80 ℃, add thermal agitation and extract 3h.After reduced pressure concentration, centrifugal acquisition clarification extract, adds 4 times of volume ethanol in extract according to 1:4 volume ratio, spends the night and staticly settles, centrifugal collecting precipitation process vacuum freeze drying after mixing at 4 ℃.By the water-soluble rear DEAE-sephadex A-25 cellulose anion-exchange column of crossing of sediment, adopt successively water and 1.0mol/L ammonium chloride solution wash-out, collect the eluent of 1.0mol/L ammonium chloride solution, adopt 3.5kD bag filter to dialyse, dialysis is dammed, and liquid passes through reduced pressure concentration and vacuum freeze drying obtains Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
Embodiment 3
A kind of Radix Et Caulis Acanthopanacis Senticosi polysaccharide that the present embodiment provides, it prepares by the following method: after wilsonii rhizome is pulverized according to solid-liquid ratio 1:20(g/mL) add ethanol, heat stirring and refluxing 7h at 80 ℃, filtration under diminished pressure, except desolventizing, reclaims medicinal material.According to solid-liquid ratio 1:10(g/mL) add water, at 70 ℃, add thermal agitation and extract 6h.After reduced pressure concentration, centrifugal acquisition clarification extract, adds 5 times of volume ethanol in extract according to 1:5 volume ratio, spends the night and staticly settles, centrifugal collecting precipitation process vacuum freeze drying after mixing at 4 ℃.By the water-soluble rear D900 type anion exchange resin of crossing of sediment, adopt successively water and 0.5mol/L liquor kalii acetici wash-out, collect the eluent of 0.5mol/L liquor kalii acetici, adopt 2.0kD bag filter to dialyse, dialysis is dammed, and liquid passes through reduced pressure concentration and vacuum freeze drying obtains Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
A β 42(amyloid-beta 42 fragments, β-amyloid peptide42, A β 42) protein aggregation, 3-nitropropionic acid, Pidolidone and paraquat are respectively the derivants of mitochondrial respiratory toxicity, toxicity of excitatory amino acid and oxidative stress toxicity, all can produce the neurotoxicity relevant to multiple the nervous system disease.Suppress the cytotoxicity of these neurotoxicity compounds and can effectively improve neural deterioration symptom undoubtedly.The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 1 is made, joins respectively in the HNB's cell SH-SY5Y with 50 μ M A β 42,7.5mM3-nitropropionic acid, 10mM Pidolidone and the modeling of 0.8mM paraquat according to 1-4mg/mL concentration.Control group adopts modeling to process, but replaces polysaccharide solution with cell culture medium, and blank group is not processed with modeling reagent and polysaccharide.At 37 ℃, hatch after 24h, add MTT solution to continue to hatch 4h.After hatching end, supernatant is abandoned in suction, adds DMSO jolting, measures light absorption value at 570nm place, and cell survival rate is the ratio of polysaccharide group light absorption value and control group light absorption value.Cell viability value represents with mean ± SD, adopts T check to compare the otherness of polysaccharide group and control group.
Figure 1A demonstration, A β 42 can significantly reduce SH-SY5Y cell viability when 50 μ M, and Radix Et Caulis Acanthopanacis Senticosi polysaccharide can improve the cell survival rate after A β 42 processes under 4mg/ml concentration, than contrast, has improved 13.6%.
Figure 1B shows, 3-nitropropionic acid can significantly reduce SH-SY5Y cell viability when 7.5mM, and Radix Et Caulis Acanthopanacis Senticosi polysaccharide all can be alleviated the neurotoxicity of 3-nitropropionic acid in 1-4mg/ml concentration range, when polysaccharide concentration is 2mg/ml, its cell survival rate has improved 7.7% than control group.
Fig. 1 C shows, Pidolidone can significantly reduce SH-SY5Y cell viability when 10mM, and Radix Et Caulis Acanthopanacis Senticosi polysaccharide shows the neurovirulent activity of inhibition Pidolidone in 1-4mg/ml concentration range, when polysaccharide concentration is 4mg/ml, its cell survival rate has improved 22.2% than control group.
Fig. 1 D shows, paraquat can significantly reduce SH-SY5Y cell viability because of oxidative damage when 0.8mM, and Radix Et Caulis Acanthopanacis Senticosi polysaccharide can suppress the neurotoxicity of paraquat in 1-4mg/ml concentration range, when polysaccharide concentration is 2mg/ml, its cell survival rate has improved 14.4% than control group.
These results show, Radix Et Caulis Acanthopanacis Senticosi polysaccharide can suppress the neurotoxicity of multiple neurotoxicity compound in human nerve cell level, shows that it has good external neuroprotection.
Embodiment 5
C. Elegans Automatic Screening transgenic models HA759 utilizes osmotic pressure related gene osm-10 to impel polyQ(poly glumine, polyglutamine, polyQ) and GFP in its head ASH neuron, express, polyQ assembles will produce remarkable toxicity, cause ASH neuron carrying out property decline fast (showing as fluorescence disappears).Therefore can come detection of drugs polyQ to be assembled to the neurovirulent mitigation of mediation by observing the neuronic survival rate of ASH.The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 2 is made, adds respectively with 1-4mg/mL concentration in the C. Elegans Automatic Screening poly glumine model HA759 larva of L1 phase, and control group adds isopyknic S Medium, is placed at 15 ℃ and cultivates 3 days.Collect HA759 C. Elegans Automatic Screening, add sodium azide solution to make its paralysis, mix rear dropping on agarose pad, at the neuronic GFP phosphor dot of every C. Elegans Automatic Screening ASH of fluorescence microscopy Microscopic observation.Random statistical～100 C. Elegans Automatic Screening, counts the neuronic survival rate of ASH.The C. Elegans Automatic Screening number of ASH neuronal survival percentage=ASH neuronal survival/(the C. Elegans Automatic Screening number of C. Elegans Automatic Screening number+ASH neuronal death of ASH neuronal survival) * 100%.Survival rate represents with mean ± SEM, uses one-way ANOVA analysis and Tukey post-hoc test to compare the otherness of polysaccharide group and control group.
As shown in Figure 2 A, the ASH neuron of the HA759 C. Elegans Automatic Screening of processing without polysaccharide becomes feeble and die (fluorescence disappearance) because polyQ assembles result, adds after Radix Et Caulis Acanthopanacis Senticosi polysaccharide ASH neuron still survive (fluorescence existence).Fig. 2 B shows, the survival rate of control group HA759 C. Elegans Automatic Screening ASH neuron after 3 days is 38.9%, and after adopting Radix Et Caulis Acanthopanacis Senticosi polysaccharide to process, the neuronic survival rate of ASH obviously improves, when polysaccharide concentration is 2mg/ml and 4mg/ml, the neuronic survival rate of ASH is increased to respectively 58.1% and 59.7% than control group.This result shows that Radix Et Caulis Acanthopanacis Senticosi polysaccharide can alleviate the neurotoxicity that polyQ assembles mediation extremely.
Therefore its height of ASH neuron control of C. Elegans Automatic Screening oozes avlidance behavior ability, is being subject to luring while stopping with hypertonic solution, and the HA759 C. Elegans Automatic Screening that ASH neuron does not become feeble and die has and tends to the ambivalent behavior feature of avoiding hypertonic solution after attractant; And the HA759 C. Elegans Automatic Screening of ASH neuronal death only has the chemotactic behavior of trend attractant, lose the avoidance ability to hypertonic solution.The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 2 is made, adds respectively with 1-4mg/mL concentration in the HA759 larva of L1 phase, and control group adds isopyknic S Medium, is placed at 15 ℃ and cultivates 3 days.Collect HA759 nematode, with PBS, wash away the swill in nutrient solution.With 8M glycerine (height oozes to detest and avoids reagent), biacetyl (luring chemotactic reagent), sodium azide (anesthetic), preparing the chemotactic of HA759 C. Elegans Automatic Screening avoids dull and stereotyped, C. Elegans Automatic Screening after processing is placed on flat board, cultivate 1.5h in 20 ℃ after, take out the distribution situation of statistics C. Elegans Automatic Screening, thereby calculate the rate of avoidance of C. Elegans Automatic Screening to hypertonic solution.Rate of avoidance (avoidance index) computing formula is as follows: rate of avoidance=(C. Elegans Automatic Screening number in region of avoidance/C. Elegans Automatic Screening sum) * 100%, rate of avoidance represents with mean ± SD, uses one-way ANOVA analysis and Tukey post-hoc test to compare the otherness of polysaccharide group and control group.
Result as shown in Figure 2 C; the control group of processing without polysaccharide has near the avoidance of turning back glycerine line of 54.5% C. Elegans Automatic Screening; after adopting Radix Et Caulis Acanthopanacis Senticosi polysaccharide to process; when polysaccharide concentration is 2mg/ml; the rate of avoidance of HA759 C. Elegans Automatic Screening is increased to 69.8%; show that Radix Et Caulis Acanthopanacis Senticosi polysaccharide has protective effect to ASH neuron, with conclusion is consistent as shown in Figure 2 A and 2 B.
Above result demonstration, Radix Et Caulis Acanthopanacis Senticosi polysaccharide can suppress the abnormal neurotoxicity causing of assembling of polyQ in neuronal cell and Animal Behavior Science level, shows that it has neuroprotection in good body.
Embodiment 6
The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 3 is made, adds with 2mg/mL concentration in the HA759 C. Elegans Automatic Screening larva of L1 phase, and control group adds isopyknic S Medium, is placed at 15 ℃ and cultivates 3 days.Collect HA759 C. Elegans Automatic Screening, add the PBS containing 1% polysorbas20, then be transferred in glass homogenizer in homogenate on ice, collect homogenate and add again DCFH-DA probe solution (indicator of active oxygen radical content), at 37 ℃, lucifuge is hatched, then with fluorescence microplate reader, detect fluorescent value, excitation wavelength is 485nm, and emission wavelength is 530nm.Relative intensity of fluorescence represents with mean ± SEM, adopts T check to compare the otherness of polysaccharide group and control group.
Result as shown in Figure 3A, the DCF relative fluorescence value of control group is 47.8, after adopting 2mg/ml Radix Et Caulis Acanthopanacis Senticosi polysaccharide to process, DCF relative fluorescence value drops to 29.1, show that Radix Et Caulis Acanthopanacis Senticosi polysaccharide can significantly reduce HA759 C. Elegans Automatic Screening activity in vivo oxygen free radicals, there is antioxidation activity in good body.
The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 3 is made, with 2mg/mL concentration, add in the L4 phase HA759 C. Elegans Automatic Screening larva of processing through 5-FUDR, with 100mM paraquat, carry out oxidative stress modeling, control group is processed equally simultaneously, but replaces polysaccharide solution with isopyknic S Medium.Be placed in 20 ℃ of cultivations, to add the time of polysaccharide and paraquat to count 0h, every the survival condition of 12h statistics C. Elegans Automatic Screening, until it is all dead, in statistic processes, health is stiff motionless and the unresponsive C. Elegans Automatic Screening of slight vibrations is judged as to dead state.C. Elegans Automatic Screening sample size 80-100 bar with each processed group is as the criterion, and C. Elegans Automatic Screening survival rate is analyzed by Kaplan-Meier method, and with log-rank test, compares the otherness of polysaccharide group and control group.
Result as shown in Figure 3 B, after adding paraquat, the acute oxidative stress of generation is caused C. Elegans Automatic Screening dead, the mean survival time of control group is 62h, and after adopting 2mg/ml Radix Et Caulis Acanthopanacis Senticosi polysaccharide to process, the mean survival time significant prolongation of C. Elegans Automatic Screening is to 71.4h, show that Radix Et Caulis Acanthopanacis Senticosi polysaccharide can effectively resist the oxidative stress of paraquat, thereby improve the oxidative damage relevant to neurological conditions.
Through above-mentioned mammalian neural cell and C. Elegans Automatic Screening disease model result of the test, confirm, Radix Et Caulis Acanthopanacis Senticosi polysaccharide not only can effectively suppress the neurotoxicity (seeing embodiment 4) of multiple neurotoxicity compound, the neurotoxicity (seeing embodiment 5) of alleviating the abnormal aggregation inducing of polyQ, can also significantly reduce disease model activity in vivo oxygen free radicals and improve the survival rate (see embodiment 6) of disease model under oxidative stress environment.These results fully prove that Radix Et Caulis Acanthopanacis Senticosi polysaccharide has good neuroprotection, can be used for preventing and treating the nervous system disease.
Embodiment 7
The Radix Et Caulis Acanthopanacis Senticosi polysaccharide that embodiment 1 is made, adds as required the preparation of appropriate lactalbumin, maltodextrin and becomes pulverulence healthy food or the health food with neuroprotective effect, can also add other conventional auxiliary material and make regular dosage form.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under Spirit Essence of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, be included in protection scope of the present invention.
Claims (9)
1. Radix Et Caulis Acanthopanacis Senticosi polysaccharide has the application in the health food of neuroprotective effect in preparation.
2. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 1 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: described neuroprotective effect refer to take protein gathering extremely, oxidative stress and the decline of specific nerve cell dead be the preventive and therapeutic effect of the nervous system disease of main pathological characters.
3. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 1 has the application in the health food of neuroprotective effect in preparation; it is characterized in that described Radix Et Caulis Acanthopanacis Senticosi polysaccharide prepares by the following method: take wilsonii as raw material; through preparing containing degreasing, water extraction, alcohol precipitation, ion-exchange and dialysis operation, described Radix Et Caulis Acanthopanacis Senticosi polysaccharide has good neuroprotection.
4. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 3 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: during degreasing, employing ethanol is degreasing solvent, and the mass volume ratio of wilsonii and ethanol is 1:2～20g/mL, skimming temp is 50～90 ℃, and the time is 1～10h; During water extraction, the mass volume ratio of wilsonii and water is 1:2～20g/mL, and extracting temperature is 50～100 ℃, and water extraction time is 1～10h; During alcohol precipitation, adopt ethanol precipitation extract, the volume ratio of ethanol and extract is 3～5:1.
5. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 3 has the application in the health food of neuroprotective effect in preparation; it is characterized in that: described ion-exchange is anion exchange; after being dissolved, alcohol hypostasis crosses anion-exchange column; adopt successively water and eluent to carry out wash-out; eluent is crossed bag filter dialysis; dislysate, through concentrated and dry processing, obtains Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
6. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 5 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: the material of described anion exchange is cellulose, sephadex, Ago-Gel or resin.
7. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 5 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: eluent is acetate, phosphate, barbiturate, citrate, sodium salt, sylvite, ammonium salt or glycine.
8. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 5 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: the molecular cut off of bag filter is 1.0～5.0kD.
9. Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 3 has the application in the health food of neuroprotective effect in preparation, it is characterized in that: described wilsonii refers to medicinal part root or the root skin of wilsonii.
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| CN107098986A (en) * | 2017-04-26 | 2017-08-29 | 江西九草铁皮石斛科技协同创新有限公司 | A kind of method that utilization low temperature stress effect improves Dendrobium officinale polysaccharide |
| CN107098986B (en) * | 2017-04-26 | 2019-06-11 | 江西九草铁皮石斛科技协同创新有限公司 | A method of it is acted on using low temperature stress and improves Dendrobium officinale polysaccharide |
| CN110407948A (en) * | 2018-04-26 | 2019-11-05 | 青岛农业大学 | A kind of extraction process of Radix Et Caulis Acanthopanacis Senticosi polysaccharide |
| CN108948221A (en) * | 2018-08-03 | 2018-12-07 | 广东药科大学 | A kind of Cultivation of Dictyophora polysaccharide and its preparation and application with neuroprotective efficacy |
| WO2022016644A1 (en) * | 2020-07-20 | 2022-01-27 | 广州青岚生物科技有限公司 | Acanthopanax senticosus harms homogeneous polysaccharide, preparation method therefor and use thereof |
| GB2613696A (en) * | 2020-07-20 | 2023-06-14 | Guangzhou Qinglan Biotechnology Co Ltd | Acanthopanax senticosus harms homogeneous polysaccharide, preparation method therefor and use thereof |
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