CN102895439A - 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 the 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 constantly worsens for cognitive and memory function, and carrying out property of activity of daily living goes down, and various neuropsychic symptoms and behavior disorder are arranged.Show according to Chinese Alzheimer association announcement survey result in 2011, the whole world 3,650 ten thousand people that have an appointment suffer from dementia, just have a people to suffer from this disease in per seven seconds, and mean survival time (MST) only has 5.9 years, is one of " four large killers " of threatening aged health.
Senile dementia mainly is divided into the dementia that the brain degenerative disease causes according to its cause of disease---and the Alzheimer dementia (is again alzheimer disease, Alzheimer ' s Disease, be called for short AD), the dementia (vascular dementia that causes of cerebrovascular, Vascular Dentia, abbreviation VaD), Mixed dementia (Mixed Dementia is called for short MixD) three major types.Secondly also comprise dull-witted (the Dementia with Lewy Bodies of Lewy body, 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), dull-witted (the Creutzfeldt-Jakob Disease of Creutzfeldt-Jakob disease, be called for short CJD), normal pressure hydrocephalus dementia (Normal Pressure Hydrocephalus is called for short NPH), dull-witted (the Wernicke-Korsakoff Syndrome of wernicke-Korsakoff syndrome, be called for short WKS), by vitamin B12, dementia due to the 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 take the intelligence infringement as main.Pathological change is mainly the cortex diffuse atrophy, and ditch returns broadening, and the ventricles of the brain enlarge, 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, and aphasia and apraxia are often arranged.Over-65s is sent out the patient and is claimed late hair style dull-witted, mostly is and distributes, and pathological development is slower, and research finds, except age of onset was had any different sooner or later, they were obviously different aspect clinical symptoms and the change of encephalopathy reason.
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, because the toxic and side effects that some chemical synthetic drugs exist in the long-term taking process, the oral administration biaavailability that peptide medicament exists is low, easily be hydrolyzed in vivo, selectivity is strong, molecular mass is large and be difficult for by shortcomings such as blood brain barrier, so that their research is faced with huge challenge.This excavates the drug research with anti-ageing year dementia activity a good chance is provided for carrying out from natural product and Chinese herbal medicine.As: therefrom the natural product huperzine A of resulting separation (huperzine A) and the galantamine that extracts from Bulbus Lycoridis Radiatae all are reversible inhibitors of acetylcholine esterase in the studies on Huperzia serrata, definite therapeutic effect is arranged, the characteristics such as have that selectivity is high, toxicity is low and effective drug duration is long are to treat in the world so far the comparatively desirable drug candidate of senile dementia.In addition, studies show that gamma-secretase is the key factor that determines that senile plaque forms, therefore present research focuses mostly in the design of inhibitors of gamma-secretase.
Rhizoma Acori Graminei is China's Chinese medicine, its anti-senile dementia bioactive molecule research on mechanism there is the important application prospect, Rhizoma Acori Graminei (Acorus tatarinowii Schott) is the 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 be fragrant, turn round and stretch out the hand perfume (or spice), sierra salvia (Sichuan) etc., grow on the other stone of wetland or small stream, mainly be 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 and comparison of the chemical composition of Rhizoma Acori Graminei deeply, and is in the majority with volatile oil part research, and the research of non-volatile position is less, mainly comprises the chemical 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 the volatile oil part, then relatively less for the research at other position.But the part modern pharmacology is the result think, nonvolatile element has good effect equally, also is one of effective site.Therefore, the real effective ingredient of Rhizoma Acori Graminei still lacks research.
Utilizing at present the Rhizoma Acori Graminei senile dementia prevention and cure mainly is by Rhizoma Acori Graminei is used, also have the extract preparation that utilizes Rhizoma Acori Graminei to treat medicine for senile dementia, for example patent CN101721567B and CN102526383A as the Herba indigoferae Pseudotinctoriae in the Chinese medicine.
The preparation method that CN101721567B discloses a kind of Rhizoma Acori Graminei extract comprises the steps: the Rhizoma Acori Graminei water extraction, extracting solution is concentrated to get 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%, drying 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 prepares in accordance with the following methods: (1) removes volatile oil: Rhizoma Acori Graminei is mixed with water, soak 1.5～2h, be heated to boiling and keep 6h, collect volatile oil, collect complete rear cooling, filter, preserve filtrate, filter cake adds water reflux, extract, 1～1.5h, cooling is filtered, and merges filtrate twice, obtain first-time filtrate, final gained filter cake is a filter cake, and with a filtration cakes torrefaction, first-time filtrate is Rhizoma Acori Graminei and removes volatile oil water extraction liquid; (2) Petroleum ether extraction: with 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 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 after repeating to extract is the secondary filter cake, the 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 extraction: with step (2) single extraction water ethyl acetate re-extract, the organic facies that merges re-extract obtains the reextraction organic facies, and final water is the reextraction water behind the re-extract; Step (2) secondary filter cake is repeated reflux, extract, with 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: with step (3) reextraction water n-butyl alcohol re-extract, the organic facies that merges with the n-butyl alcohol re-extract obtains three extracted organic phase, is three aqueous phase extracted with the final remaining water of n-butyl alcohol re-extract; (3) three filter cakes of step are repeated 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 concentrating under reduced pressure with four filtrates and three extracted organic phase, obtain concentrate, i.e. n-butanol extract; (5) ethanol extraction: (4) four filter cakes of step are repeated reflux, extract, with ethanol, filter, merging repeats the filtrate that reflux, extract, obtains with ethanol, be five filtrates, the filter cake that repeats finally to obtain after the reflux, extract, with ethanol is five filter cakes, and five filtration cakes torrefactions are concentrated with five filtrate decompression, obtain concentrate, i.e. ethanol extraction; (6) water extract: with (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 extract on senile dementia prevention and cure of the disclosed Rhizoma Acori Graminei of these patents is unsatisfactory, and possible cause is to be limited to the extracting method of effective ingredient in the 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 the 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 the senile dementia prevention and cure medicine, by great many of experiments, obtained to obtain to have for senile dementia extracting method and the effective dose in the treatment senile dementia of prevention effect.
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, the second portion extract composition that this extract obtains first's extract and residue passed through organic solvent extraction by Rhizoma Acori Graminei being carried out 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 are petroleum ether, ethyl acetate and n-butyl alcohol.
Preferably, in the above-mentioned Rhizoma Acori Graminei extract, the preparation method of first's extract is as follows:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain first's Rhizoma Acori Graminei extract with the described desorbed solution of hcl acidifying.
Preferably, in the above-mentioned Rhizoma Acori Graminei extract, the preparation method of second portion extract is as follows:
(3) residue that will obtain in the time of will obtaining first's extract uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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 the second portion 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 the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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 places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 5 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, by 001 * 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=6, at 50 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol 20 ℃ of lower lixiviates 3 hours 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 places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 10 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, and by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=8, at 60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 30 ℃ of lower lixiviate 3-4 hours 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 the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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 the senile dementia prevention and cure disease, the 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.
The specific embodiment
In order to understand the present invention, the below further specifies the present invention with embodiment, but does 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 places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 5 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, by 001 * 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=6, at 50 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol 20 ℃ of lower lixiviates 3 hours 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 places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 10 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, and by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=8, at 60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 30 ℃ of lower lixiviate 3-4 hours 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's 1 contrast Rhizoma Acori Graminei extract 1
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized places the 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 gets the Rhizoma Acori Graminei water extract at 50 ℃ of lower concentrating under reduced pressure;
(2) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 30 ℃ of lower lixiviate 3-4 hours 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 the 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's 2 contrast Rhizoma Acori Graminei extracts 2
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 5 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, by 001 * 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=6, at 50 ℃ of concentrating under reduced pressure, obtain contrast Rhizoma Acori Graminei extract 2 with the described desorbed solution of hcl acidifying;
EXPERIMENTAL EXAMPLE 1
Whether can regulate A β level in order to test Rhizoma Acori Graminei extract, sandwich enzyme-linked immunosorbent assay (ELISA) is used 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 the 6 hole flat boards with 200,000 cells/well and 37 ℃ 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 the 1.5ml culture medium of 10mg/l, 50mg/l, 100mg/l, 150mg/l, 200mg/l, 250mg/l concentration, with the medium treatment cell of preparation 48 hours.To be collected into Eppendorf tube and analyze from the cell conditioned medium liquid of processing.Test each extract to the impact of A β 42 in the H4 cell and A β 40 content, A β 42 and A β 40 changes of contents ratios behind described numeric representation and the standard control.
The extract of table 1: embodiment 1 and embodiment 2 is on the impact of A β 42 in the H4 cell
The extract of table 2: embodiment 1 and embodiment 2 is on the impact of A β 40 in the H4 cell
Table 1 and table 2 show that the Rhizoma Acori Graminei extract by embodiment 1 and embodiment 2 preparations can reduce A β 42 in the H4 cell and the content of A β 40 effectively.Comparative example 1 and 2 then effect is bad.
Simultaneously the APP amyloid is crossed stable cell line H4 wasted bottle detection embodiment 1 and 2 extracts of expression, obvious reflection is arranged.
Embodiments of the invention 1 and 2 extract can be realized the good result to the senile dementia cell model.
EXPERIMENTAL EXAMPLE 2
With 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, the 18-22 gram is according to the body weight random packet.Guard time to memory is averaged, and numerical value unit is second.
Table 3 Rhizoma Acori Graminei extract is to the experimental result of dysmnesia mice
Time the 14th day, the test result of medicine group and model group be significant difference (p＜0.01) relatively
The result shows, the Rhizoma Acori Graminei extract of embodiment 1 has significant protective effect to obtaining the dysmnesia mice.Method is different, and comparative example 1 Rhizoma Acori Graminei extract is not obvious in the amnemonic protective effect to mice.
Claims (7)
1. Rhizoma Acori Graminei extract for the treatment of senile dementia, the second portion extract composition that this extract obtains first's extract and residue passed through organic solvent extraction by Rhizoma Acori Graminei being carried out 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 are petroleum ether, ethyl acetate and n-butyl alcohol.
2. Rhizoma Acori Graminei extract according to claim 1, wherein the preparation method of first's extract is as follows:
(1) the Rhizoma Acori Graminei leaf powder of pulverizing is placed the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain first's Rhizoma Acori Graminei extract with the described desorbed solution of hcl acidifying.
3. Rhizoma Acori Graminei extract according to claim 1, wherein the preparation method of second portion extract is as follows:
(3) residue that will obtain in the time of will obtaining first's extract uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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 the second portion extract.
4. 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 the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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.
5. Rhizoma Acori Graminei extract for the treatment of senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 5 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, by 001 * 7 cation exchange resin, the sodium hydrate aqueous solution with 4% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=6, at 50 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol 20 ℃ of lower lixiviates 3 hours 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.
6. Rhizoma Acori Graminei extract for the treatment of senile dementia, this extract method as follows obtains:
(1) the Rhizoma Acori Graminei leaf powder of 100g being pulverized places the 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 the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 50 ℃ of lower concentrating under reduced pressure;
(2) water that adds 10 times of crude extract weight in the Rhizoma Acori Graminei crude extract stirs into the crude extract suspension, and by cation exchange resin, the sodium hydrate aqueous solution with 8% is resolved the acquisition desorbed solution to cation exchange resin with this suspension; To pH=8, at 60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 30 ℃ of lower lixiviate 3-4 hours 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.
7. 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 the supersound extraction device, the weight ratio of Rhizoma Acori Graminei and water is 1: 30-50, soak 100-150min, supersonic frequency 30-40kHz, extract 50-60 ℃ of temperature, extraction time 110-170min, the extracting solution after the supersound extraction carries out sucking filtration, obtain clear liquid and residue, clear liquid gets the Rhizoma Acori Graminei crude extract at 40-50 ℃ of lower concentrating under reduced pressure;
(2) in the Rhizoma Acori Graminei crude extract, add crude extract weight 5-10 water doubly and stir into the crude extract suspension, this suspension by cation exchange resin, is resolved the acquisition desorbed solution with the sodium hydrate aqueous solution of 4-8% to cation exchange resin; To pH=6-8, at 50-60 ℃ of concentrating under reduced pressure, obtain Rhizoma Acori Graminei extract I with the described desorbed solution of hcl acidifying;
(3) residue that (1) step was obtained uses petroleum ether, ethyl acetate and n-butyl alcohol at 20-30 ℃ of lower lixiviate 3-4 hour 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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| 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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