CN102895439B - Rhizoma acori graminei extract for treating Alzheimer and method for extracting rhizoma acori graminei extract for treating Alzheimer - Google Patents
Rhizoma acori graminei extract for treating Alzheimer and method for extracting rhizoma acori graminei extract for treating Alzheimer Download PDFInfo
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Abstract
The invention provides a rhizoma acori graminei extract for treating Alzheimer and a method for extracting the rhizoma acori graminei extract for treating Alzheimer. The extract consists of a first part of extract and a second part of extract, wherein the first part of extract is obtained through the steps of the ultrasonical extracting and cation exchange resin analyzing of rhizoma acori graminei, the second part of extract is obtained through extracting residues through organic solvent, water is used as the solvent for ultrasonic extraction, the ultrasonic frequency is 30-40kHz, sodium hydroxide aqueous solution is adopted for the analyzing of a cation exchange resin column, hydrochloric acid used as analyzing liquid for analyzing, and petroleum ether, ethyl acetate and n-butyl alcohol are used as the organic solvent. The rhizoma acori graminei extract for treating Alzheimer has an excellent effect and an obvious Alzheimer prevention effect. Compared with other conventional methods, the method for extracting the rhizoma acori graminei extract also has an excellent effect.
Description
Technical field
The invention belongs to medicine preparation field, particularly relate to a kind of Rhizoma Acori Graminei extract and extracting method thereof for the treatment of dementia.
Background technology
Alzheimer is so-called senile dementia.Be a kind of lethal neurodegenerative diseases that carries out sexual development, clinical manifestation is cognitive and memory function constantly worsens, and carrying out property of activity of daily living goes down, and has various neuropsychic symptoms and behavior disorder.Show according to the Chinese Alzheimer association announcement survey result of 2011, the whole world 3,650 ten thousand people that have an appointment suffer from dementia, within every seven seconds, just have a people to suffer from this disease, and mean survival time (MST) only has 5.9 years, is one of " four large killers " of threatening aged health.
Senile dementia is mainly divided into according to its cause of disease the dementia that brain degenerative disease causes---and Alzheimer dementia (is again alzheimer disease, Alzheimer ' s Disease, be called for short AD), the dementia (vascular dementia that causes of cerebrovascular, Vascular Dentia, be called for short VaD), Mixed dementia (Mixed Dementia, be called for short MixD) three major types.Secondly also comprise Lewy body dementia (Dementia with Lewy Bodies, be called for short DLB), Frontotemporal dementia (Frontotemporal Dementia, be called for short FTD), dementia in Parkinson disease (Parkinson ' s Disease, be called for short PD), Creutzfeldt-Jakob disease dementia (Creutzfeldt-Jakob Disease, be called for short CJD), normal pressure hydrocephalus dementia (Normal Pressure Hydrocephalus, be called for short NPH), wernicke-Korsakoff syndrome dementia (Wernicke-Korsakoff Syndrome, be called for short WKS), by vitamin B12, dementia due to folic acid deficiency etc.
The a lot of diseases of senile dementia are in the geratic period, the onset of hiding, and the course of disease is slow and irreversible, clinically taking intelligence infringement as main.Pathological change is mainly cortex diffuse atrophy, and ditch returns broadening, and the ventricles of the brain expand, and neuron reduces in a large number, and visible senile plaque, the pathological changes such as neurofibril knot, and choline acetylase and acetyl choline content significantly reduce.It is dull-witted early stage that onset was once called as senile disease at 65 years old with the former, or presenile dementia, has with sick family history more, and PD is very fast, and temporal lobe and parietal lobe lesion are more remarkable, often have aphasia and apraxia.Over-65s is sent out patient and is claimed late hair style dementia, mostly is and distributes, and pathological development is slower, research discovery, and except age of onset is had any different sooner or later, they are obviously different aspect clinical symptoms and brain pathological change.
The medicine for the treatment of senile dementia mainly contains cholinesterase inhibitor at present, cerebral vasodilator, and calcium antagonists, prevents A β deposition medicine, alpha-secretase enzyme agonist, beta-secretase inhibitor, inhibitors of gamma-secretase and adjusting control agent etc.These medicines overwhelming majority is chemical synthetic drug or peptide medicament, the toxic and side effects existing in long-term taking process due to some chemical synthetic drugs, oral administration biaavailability that peptide medicament exists is low, be easily hydrolyzed in vivo, selectivity is strong, molecular mass is compared with large and be difficult for by shortcomings such as blood brain barrier, makes their research be faced with huge challenge.This excavates the drug research with anti-alzheimer disease activity a good chance is provided for carrying out from natural product and Chinese herbal medicine.As: therefrom in studies on Huperzia serrata, the natural product huperzine A of resulting separation (huperzine A) and the galantamine that extracts from Bulbus Lycoridis Radiatae are all reversible inhibitors of acetylcholine esterase, there is definite therapeutic effect, the feature such as have that selectivity is high, toxicity is low and effective drug duration is long is to treat in the world so far the comparatively desirable drug candidate of senile dementia.In addition, research shows that gamma-secretase is the key factor that determines that senile plaque forms, and therefore present research focuses mostly in the design of inhibitors of gamma-secretase.
Rhizoma Acori Graminei is China's Chinese medicine, research to its anti-senile dementia bioactive molecule mechanism has important application prospect, Rhizoma Acori Graminei (Acorus tatarinowii Schott) is Acorus of Araceae plant, different name Rhizoma Anemones Altaicae (Jiangsu, Zhejiang, Jiangxi, Hunan), purple ear, stone Scolopendra, smelly Chang (Guangdong), Herba Kyllingae, wild leek, Rhizoma et radix valerianae (Yunnan), night perfume (or spice), turn round and stretch out the hand perfume (or spice), sierra salvia (Sichuan) etc., grow on the other stone of wetland or small stream, be mainly distributed in each provinces and regions on the south China the Yellow River.Its rhizome is used as medicine, and has removing dampness by means of aromatics, a function of the eliminating phlegm of having one's ideas straightened out, refreshment Fructus Alpiniae Oxyphyllae, the clinical diseases such as forgetful, apoplectic aphasia, tinnitus, alzheimer disease that are widely used in.The research of the chemical composition of Rhizoma Acori Graminei is more deep, in the majority with volatile oil part research, and the research of non-volatile position is less, mainly comprises the compounds such as Phenylpropanoid Glycosides class, terpenoid, alkaloids, aldehydic acid class, quinone ketone, sterols, amino acids and saccharide.A large amount of pharmaceutical researches show, Rhizoma Acori Graminei all has regulating action to multiple systems such as nervus centralis, cardiovascular, breathing and digestion, but the material base of its performance drug action is never clear and definite, think that for many years the main effective ingredient of Rhizoma Acori Graminei is volatile oil, therefore its research is also concentrated on to volatile oil part, relatively less for the research at other position.But part modern pharmacology result thinks, nonvolatile element has good effect equally, is also one of effective site.Therefore, the real effective ingredient of Rhizoma Acori Graminei still lacks research.
Utilizing at present Rhizoma Acori Graminei senile dementia prevention and cure is mainly by using Rhizoma Acori Graminei, the Herba indigoferae Pseudotinctoriae in Chinese medicine uses, and also has the extract preparation treatment medicine for senile dementia that utilizes Rhizoma Acori Graminei, for example patent CN101721567B and CN102526383A.
The preparation method that CN101721567B discloses a kind of Rhizoma Acori Graminei extract comprises the steps: Rhizoma Acori Graminei water extraction, extracting solution is concentrated to obtain to concentrated solution, go up afterwards nonpolarity macroporous adsorptive resins chromatographic column, gradient elution, gradient is from water to 35% ethanol water, the eluent of the ethanol water of collection 15～35%, dry, can make Rhizoma Acori Graminei extract; Percentage ratio is percent by volume.
CN102526383A discloses a kind of Rhizoma Acori Graminei water extract, it is characterized in that described Rhizoma Acori Graminei water extract prepared in accordance with the following methods: (1) removes volatile oil: Rhizoma Acori Graminei is mixed with water, soak 1.5～2h, be heated to seethe with excitement and keep 6h, collect volatile oil, cooling after collection, filter, preserve filtrate, the filter cake reflux, extract, 1～1.5h that adds water, cooling, filter, merge twice filtrate, obtain first-time filtrate, final gained filter cake is a filter cake, and by a filtration cakes torrefaction, first-time filtrate is Rhizoma Acori Graminei and removes volatile oil water extraction liquid; (2) Petroleum ether extraction: by step (1) first-time filtrate petroleum ether re-extract, merge respectively organic facies and water, obtain single extraction organic facies and single extraction water; (1) filter cake of step is soaked to 1～2h with petroleum ether, reflux, extract, 1～1.5h, repeats to extract, and filters, merge and repeat to extract filtration gained filtrate, obtain secondary filtrate, the filter cake repeating after extracting is secondary filter cake, secondary filtration cakes torrefaction, secondary filtrate and single extraction organic facies are merged, concentrating under reduced pressure, obtains concentrate, i.e. ligroin extraction; (3) ethyl acetate is extracted: by step (2) single extraction water ethyl acetate re-extract, the organic facies that merges re-extract obtains reextraction organic facies, and after re-extract, final water is reextraction water; Step (2) secondary filter cake is repeated to reflux, extract, by ethyl acetate, filter, merge and repeat reflux, extract, filtrate, obtain three times filtrate, repeating the final filter cake of reflux, extract, is three filter cakes, three filtration cakes torrefactions, three filtrate and reextraction organic facies are merged, concentrating under reduced pressure, obtains concentrate, i.e. ethyl acetate extract; (4) n-butanol extraction: by step (3) reextraction water n-butyl alcohol re-extract, merge and obtain three extracted organic phase by the organic facies of n-butyl alcohol re-extract, with the water of the final remainder of n-butyl alcohol re-extract be three aqueous phase extracted; (3) three filter cakes of step are repeated to reflux, extract, with n-butyl alcohol, filter, the filtrate of repeating reflux, extract, with n-butyl alcohol is four filtrates, repeating the final remaining filter cake of reflux, extract, with n-butyl alcohol is four filter cakes, four filtration cakes torrefactions, merge four filtrates and three extracted organic phase concentrating under reduced pressure, obtain concentrate, i.e. n-butanol extract; (5) ethanol extraction: (4) four filter cakes of step are repeated to reflux, extract, with ethanol, filter, merge the filtrate obtaining by ethanol repetition reflux, extract,, be five filtrates, with the filter cake that ethanol repeats finally to obtain after reflux, extract, be five filter cakes, five filtration cakes torrefactions, by concentrated five filtrate decompression, obtain concentrate, i.e. ethanol extraction; (6) water extract: by (4) three aqueous phase extracted concentrating under reduced pressure of step, obtain concentrate, i.e. Rhizoma Acori Graminei water extract, the further separation and purification of described Rhizoma Acori Graminei water extract obtains glucosides and alkaloid.
The effect of the extract of the disclosed Rhizoma Acori Graminei of these patents in senile dementia prevention and cure is unsatisfactory, and possible cause is to be limited to the extracting method of effective ingredient in Rhizoma Acori Graminei unreasonablely, causes effective ingredient not extracted as much as possible.
Because the extract of Rhizoma Acori Graminei is mixture, so extracting method of its active and Rhizoma Acori Graminei and relevant, for obtain of the present invention can senile dementia prevention and cure medicine, the present invention is by a large amount of screenings, find that Rhizoma Acori Graminei has excellent effect as the plant of extracting senile dementia prevention and cure medicine, by great many of experiments, obtain can obtain for senile dementia and there is the extracting method of prevention effect and the effective dose in senile dementia in treatment.
Summary of the invention
For solving the problems of the technologies described above, the invention provides a kind of Rhizoma Acori Graminei extract for the treatment of senile dementia and uses thereof.
Concrete, the invention provides a kind of Rhizoma Acori Graminei extract for the treatment of senile dementia, this extract obtains Part I extract and residue is consisted of the Part II extract of organic solvent extraction by Rhizoma Acori Graminei being carried out to supersound extraction and cation exchange resin analyzing step, wherein the solvent of supersound extraction is water, supersonic frequency is 30-40kHz, cation is changed the parsing sodium hydrate aqueous solution of resin column, desorbed solution resolving hydrochloric acid, organic solvent is petroleum ether, ethyl acetate and n-butyl alcohol.
Preferably, in above-mentioned Rhizoma Acori Graminei extract, the preparation method of Part I extract is as follows:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Part I Rhizoma Acori Graminei extract with desorbed solution described in hcl acidifying.
Preferably, in above-mentioned Rhizoma Acori Graminei extract, the preparation method of Part II extract is as follows:
(3) residue obtaining when obtaining Part I extract is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Part II extract;
Concrete, the invention provides a kind of Rhizoma Acori Graminei extract for the treatment of senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract mixture of senile dementia prevention and cure disease.
Preferably, described Rhizoma Acori Graminei extract method as follows prepares:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30, soak 100min, supersonic frequency 30kHz, extract temperature 50 C, extraction time 110min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 5 times of crude extract weight to stir into crude extract suspension, by this suspension, by 001 × 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved and is obtained desorbed solution cation exchange resin; To pH=6, at 50 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used the lixiviate 3 hours at 20 DEG C of petroleum ether, ethyl acetate and n-butyl alcohol successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract of senile dementia prevention and cure disease.
Preferably, described Rhizoma Acori Graminei extract method as follows prepares:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 50, soak 150min, supersonic frequency 40kHz, extract temperature 60 C, extraction time 170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 10 times of crude extract weight to stir into crude extract suspension, by this suspension, by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved and is obtained desorbed solution cation exchange resin; To pH=8, at 60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:10ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract of senile dementia prevention and cure disease.
The present invention also provides a kind of method of extracting medicine that can senile dementia prevention and cure from Rhizoma Acori Graminei, and the method comprises the steps:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the medicine of senile dementia prevention and cure.
The present invention also provides the purposes of above-mentioned Rhizoma Acori Graminei extract mixture for the preparation of the medicine of senile dementia prevention and cure disease.
Rhizoma Acori Graminei extract provided by the invention has excellent effect as senile dementia prevention and cure disease, senile dementia prevention and cure successful, and the method for the acquisition of this extract is compared to other conventional methods and also has excellent effect.
Detailed description of the invention
In order to understand the present invention, further illustrate the present invention with embodiment below, but do not limit the present invention.
The preparation of embodiment 1 Rhizoma Acori Graminei extract 1
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30, soak 100min, supersonic frequency 30kHz, extract temperature 50 C, extraction time 110min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 5 times of crude extract weight to stir into crude extract suspension, by this suspension, by 001 × 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved and is obtained desorbed solution cation exchange resin; To pH=6, at 50 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used the lixiviate 3 hours at 20 DEG C of petroleum ether, ethyl acetate and n-butyl alcohol successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II; ;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract 1 of senile dementia prevention and cure disease.
The preparation of embodiment 2 Rhizoma Acori Graminei extracts 2
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 50, soak 150min, supersonic frequency 40kHz, extract temperature 60 C, extraction time 170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 10 times of crude extract weight to stir into crude extract suspension, by this suspension, by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved and is obtained desorbed solution cation exchange resin; To pH=8, at 60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:10ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract 2 of senile dementia prevention and cure disease.
Comparative example 1 contrasts Rhizoma Acori Graminei extract 1
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in distillation extraction device, and the weight ratio of Rhizoma Acori Graminei and water is 1: 50, and extraction time 170min obtains distillate and residue after extracting, and distillate concentrating under reduced pressure at 50 DEG C obtains Rhizoma Acori Graminei water extract;
(2) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:10ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei organic extract;
(4) merge Rhizoma Acori Graminei water extract and Rhizoma Acori Graminei organic extract, obtain the contrast Rhizoma Acori Graminei extract 1 of senile dementia prevention and cure disease.
Comparative example 2 contrasts Rhizoma Acori Graminei extract 2
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30, soak 100min, supersonic frequency 30kHz, extract temperature 50 C, extraction time 110min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 5 times of crude extract weight to stir into crude extract suspension, by this suspension, by 001 × 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved and is obtained desorbed solution cation exchange resin; To pH=6, at 50 DEG C of concentrating under reduced pressure, obtain contrast Rhizoma Acori Graminei extract 2 with desorbed solution described in hcl acidifying;
EXPERIMENTAL EXAMPLE 1
Whether can regulate A β level in order to test Rhizoma Acori Graminei extract, by sandwich enzyme-linked immunosorbent assay (ELISA) for measuring A β (A β 42 and/or the A β 40) level of secretion.In the present embodiment, the H4 cell of expressing wild type APP695 is seeded on 6 hole flat boards with 200,000 cells/well and 37 DEG C with 5% carbon dioxide under overnight incubation.Respectively the 1-2 of the contrast Rhizoma Acori Graminei extract of the Rhizoma Acori Graminei extract 1 and 2 of embodiment 1 and 2, comparative example 1-2 is mixed with to the 1.5ml culture medium of 10mg/l, 50mg/l, 100mg/l, 150mg/l, 200mg/l, 250mg/l concentration, the medium treatment cell of use preparation 48 hours.To be collected into Eppendorf tube and analyze from the cell conditioned medium liquid of processing.Test the impact of each extract on A β 42 in H4 cell and A β 40 content, A β 42 and A β 40 changes of contents ratios after described numeric representation and standard control.
The impact of the extract of table 1: embodiment 1 and embodiment 2 on A β 42 in H4 cell
The impact of the extract of table 2: embodiment 1 and embodiment 2 on A β 40 in H4 cell
Table 1 and table 2 show that the Rhizoma Acori Graminei extract of preparing by embodiment 1 and embodiment 2 can reduce A β 42 in H4 cell and the content of A β 40 effectively.Effect is bad for comparative example 1 and 2.
Stable cell line H4 wasted bottle detection embodiment 1 and 2 extracts of simultaneously APP amyloid being crossed expression, have obvious reflection.
The extract of embodiments of the invention 1 and 2 can be realized the good result to senile dementia cell model.
EXPERIMENTAL EXAMPLE 2
By normal group and model group gavage every day normal saline 0.2ml/10g, the extracting solution 0.2ml/10g of medicine group 1 gastric infusion embodiment 1.Medicine group 2 gastric infusion comparative examples' 1 extracting solution 0.2ml/10g, normal group is normal mouse, and model group and medicine group 1 and 2 are to obtain amnemonic mice, and 20 every group, 18-22 gram, according to body weight random packet.Guard time to memory is averaged, and numerical value unit is second.
The experimental result of table 3 Rhizoma Acori Graminei extract to dysmnesia mice
Time the 14th day, the test result of medicine group and model group be significant difference (p<0.01) relatively
Result shows, the Rhizoma Acori Graminei extract of embodiment 1 has significant protective effect to obtaining dysmnesia mice.Method difference, comparative example 1 Rhizoma Acori Graminei extract is not obvious in the amnemonic protective effect to mice.
Claims (6)
1. treat the Rhizoma Acori Graminei extract of senile dementia for one kind, this extract obtains Part I extract and residue is consisted of the Part II extract of organic solvent extraction by Rhizoma Acori Graminei being carried out to supersound extraction and cation exchange resin analyzing step, wherein the solvent of supersound extraction is water, supersonic frequency is 30-40kHz, cation is changed the parsing sodium hydrate aqueous solution of resin column, desorbed solution resolving hydrochloric acid, organic solvent is petroleum ether, ethyl acetate and n-butyl alcohol; Wherein the preparation method of Part I extract is as follows:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension, by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Part I Rhizoma Acori Graminei extract with desorbed solution described in hcl acidifying.
2. Rhizoma Acori Graminei extract according to claim 1, wherein the preparation method of Part II extract is as follows:
(3) residue obtaining when obtaining Part I extract is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Part II extract;
3. treat a Rhizoma Acori Graminei extract for senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension, by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract of senile dementia prevention and cure disease.
4. treat a Rhizoma Acori Graminei extract for senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30, soak 100min, supersonic frequency 30kHz, extract temperature 50 C, extraction time 110min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 5 times of crude extract weight to stir into crude extract suspension, by this suspension, by 001 × 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved and is obtained desorbed solution cation exchange resin; To pH=6, at 50 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used the lixiviate 3 hours at 20 DEG C of petroleum ether, ethyl acetate and n-butyl alcohol successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract of senile dementia prevention and cure disease.
5. treat a Rhizoma Acori Graminei extract for senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized is placed in supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 50, soak 150min, supersonic frequency 40kHz, extract temperature 60 C, extraction time 170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add the water of 10 times of crude extract weight to stir into crude extract suspension, by this suspension, by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved and is obtained desorbed solution cation exchange resin; To pH=8, at 60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:10ml, and the extract of petroleum ether, ethyl acetate and n-butyl alcohol merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the Rhizoma Acori Graminei extract of senile dementia prevention and cure disease.
6. a method of extracting medicine that can senile dementia prevention and cure from Rhizoma Acori Graminei, the method comprises the steps:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed in to supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract temperature 50-60 DEG C, extraction time 110-170min, the extracting solution after supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid concentrating under reduced pressure at 40-50 DEG C obtains Rhizoma Acori Graminei crude extract;
(2) in Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly to stir into crude extract suspension, this suspension, by cation exchange resin, is resolved to acquisition desorbed solution to cation exchange resin with the sodium hydrate aqueous solution of 4-8%; To pH=6-8, at 50-60 DEG C of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with desorbed solution described in hcl acidifying;
(3) residue (1) step being obtained is used petroleum ether, ethyl acetate and n-butyl alcohol lixiviate 3-4 hour at 20-30 DEG C successively, the w/v of residue and petroleum ether, ethyl acetate and n-butyl alcohol is 1g:5-10ml, and extract merges concentrating under reduced pressure and obtains Rhizoma Acori Graminei extract II;
(4) merge Rhizoma Acori Graminei extract I and Rhizoma Acori Graminei extract II, obtain the medicine of senile dementia prevention and cure.
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| CN101085129A (en) * | 2007-07-11 | 2007-12-12 | 石任兵 | Acorus gramineus total phenylpropanoid extraction and total phenols extraction and method for preparing simultaneously |
| CN101721567A (en) * | 2008-10-10 | 2010-06-09 | 上海医药工业研究院 | Grassleaf sweetflag rhizome extract, preparation method and application thereof |
| CN101897843A (en) * | 2009-06-01 | 2010-12-01 | 王登之 | New preparation of effective fractions of rhizoma acori graminei and preparation method thereof |
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| CN101721567A (en) * | 2008-10-10 | 2010-06-09 | 上海医药工业研究院 | Grassleaf sweetflag rhizome extract, preparation method and application thereof |
| CN101897843A (en) * | 2009-06-01 | 2010-12-01 | 王登之 | New preparation of effective fractions of rhizoma acori graminei and preparation method thereof |
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