CN103285071B - Medical use of milkvetch root total saponin - Google Patents
Medical use of milkvetch root total saponin Download PDFInfo
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Abstract
The invention relates to the field of medicine, particularly to a medical use, a drug composition and a health product of milkvetch root total saponin. The milkvetch root total saponin can be used for preparing a drug or a health product for improving memory of normal people.
Description
Technical field
The present invention relates to Medicines and Health Product field, particularly relate to a kind of medical usage and pharmaceutical composition and health product of Radix Astragali total saponins.
Background technology
The knowledge of obtaining by study must be stored in some way, and reads out over time, just can cause the change of individual behavior.This " access " process is exactly our said memory.According to asynchronism(-nization) required between access, people are divided into roughly impermanent memory and longterm memory memory conventionally.The time scale of impermanent memory (claiming again working memory) is generally the several seconds to arrive several minutes, the cognitive process restriction such as is subject to attention.Impermanent memory generally can only store a small amount of information.Under specific circumstances, the information of impermanent memory system storage can be transferred to longterm memory system, and is strengthened and consolidate at this, reach several hours, several days even lifelong.Longterm memory is divided into again declarative memory and procedural memory, and reference memory belongs to declarative memory.
Radix Astragali total saponins is one of active component of the Radix Astragali, and it comprises astragaloside I, II, III, IV, V, VI, VII, different astragaloside I and II and Radix Astragali saponin unit etc., it has adjusting blood glucose, enhancing human body immunity power, promotes growth, improves the effects such as antioxidant ability of organism.Research shows, Radix Astragali total saponins can obviously suppress the hypertrophy of neutrophilic granulocyte and reduce the cytokine of inflammation local joint, as interleukin-1 and nitric oxide production level, alleviate articular chondrocyte apoptosis that nitric oxide causes etc., and toxicity is low, and side effect is little.Have no so far the report that can improve normal mouse memory about Radix Astragali total saponins.
Summary of the invention
Object of the present invention aims to provide a kind of new medical usage and pharmaceutical composition and health product of Radix Astragali total saponins.
Specifically, first aspect present invention has been to provide a kind of Radix Astragali total saponins in the medicine of preparation raising memory in normal people or the application in health product.
In a preference, in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
In another preference, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
In another preference, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
A second aspect of the present invention has been to provide a kind of pharmaceutical composition that improves memory in normal people, and it comprises the Radix Astragali total saponins for the treatment of effective dose, and in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
In a preference, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
In another preference, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
The present invention also provides a kind of health product that improve memory in normal people, the Radix Astragali total saponins that it comprises effective dose, and in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
In a preference, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
In another preference, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
The details of various aspects of the present invention will be able to detailed description in chapters and sections subsequently.By below and the description of claim, feature of the present invention, object and advantage will be more obvious.
Accompanying drawing explanation
Fig. 1 has embodied the content (ELSD figure) of astragaloside in Radix Astragali total saponins of the present invention, and wherein the appearance time of astragaloside is 11.66min;
Fig. 2 A has embodied the impact of Radix Astragali total saponins of the present invention on normal KM mice eight arm maze experiment reference memory mistakes.（*,p<0.05）；
Fig. 2 B has embodied the impact of Radix Astragali total saponins of the present invention on the first wrong front positive exact figures of normal KM mice eight arm maze experiments.（*,p<0.05）；
Fig. 3 has embodied Radix Astragali total saponins of the present invention normal KM mice is shuttled back and forth in experiment and initiatively escapes the impact of number of times.（*，p<0.05；+，p<0.07）。
The specific embodiment
Appearance part of the present invention is based on so unexpected discovery: Radix Astragali total saponins can significantly improve the memory of normal mouse.Therefore, Radix Astragali total saponins can be used for medicine or the health product that preparation improves memory in normal people.
And then a first aspect of the present invention has been to provide Radix Astragali total saponins and has improved the medicine of memory in normal people or the application in health product in preparation.
Preferably, in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
More preferably, in described Radix Astragali total saponins the content of astragaloside lower than 1%.
Most preferably, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1:1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
A second aspect of the present invention has been to provide a kind of pharmaceutical composition that improves memory in normal people, and it comprises the Radix Astragali total saponins for the treatment of effective dose, and in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
Preferably, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
More preferably, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
The present invention also provides a kind of health product that improve memory in normal people, the Radix Astragali total saponins that it comprises effective dose, and in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
Preferably, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
More preferably, described Radix Astragali total saponins can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
As known for one of ordinary skill in the art, Radix Astragali total saponins of the present invention can be by the conventional method of this area as water extraction, alcohol extracting method, extraction are refined, resin adsorption method is refining and supercritical CO
2methods etc. are extracted and are obtained from the Radix Astragali, also can be by commercial sources buy from Sigma chemical company, Man Site bio tech ltd, Chengdu etc. and obtain.Its purity all meets medicinal standard.Preferably, in Radix Astragali total saponins of the present invention the content of astragaloside lower than 5%; More preferably, in Radix Astragali total saponins of the present invention the content of astragaloside lower than 1%; Most preferably, Radix Astragali total saponins of the present invention can make by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by water saturated n-butyl alcohol for extracting solution (1: the 1) extraction after above-mentioned concentrating 2～4 times, combining extraction liquid, reclaims n-butyl alcohol, concentrated;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d), by above-mentioned crude extract dissolve with methanol, then add the ether of 3～5 times of amounts: acetone (1:1) solution, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
Radix Astragali total saponins of the present invention can be used separately or use with the form of pharmaceutical composition.Pharmaceutical composition comprises Radix Astragali total saponins of the present invention and the pharmaceutically suitable carrier as active component.Preferably, the Radix Astragali total saponins of the present invention as active component that pharmaceutical composition of the present invention contains 0.1-99.9% percentage by weight." pharmaceutically suitable carrier " can not destroy the pharmaceutical active of Radix Astragali total saponins of the present invention, its effective dose simultaneously, and can bringing into play pharmaceutical carrier, to make the consumption of used time nontoxic to human body.
Described pharmaceutically suitable carrier includes but not limited to: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance are if phosphate, glycine, sorbic acid, water, salt, electrolyte are as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate, satisfied fatty acid partial glycerol ester admixture etc.
Other conventional excipient substances are as binding agent (as microcrystalline Cellulose), filler (as starch, glucose, Lactis Anhydrous and lactose beadlet), disintegrating agent (as cross-linked pvp, crosslinked carboxymethyl fecula sodium, cross-linking sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose), lubricant (as magnesium stearate) and absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent, wetting agent etc.
Radix Astragali total saponins of the present invention with and pharmaceutical composition can and can pass through intestinal or non-intestinal or topical routes by this area conventional method preparation.Oral formulations comprises capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.; Non-intestinal drug delivery agent comprises injection etc.; Local administration preparation comprises cream, patch, ointment, spray etc.Be preferably oral formulations.
Radix Astragali total saponins of the present invention take and the route of administration of pharmaceutical composition can be as oral, Sublingual, percutaneous, through muscle or subcutaneous, mucocutaneous, vein, urethra, vagina etc.
Except making medicament, also can in Radix Astragali total saponins of the present invention, add the various food additive such as antioxidant, pigment, enzyme preparation, by the conventional method of this area, make health food.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percent, ratio, ratio or umber by weight.
Unless otherwise defined, the same meaning that all specialties of using in literary composition and scientific words and one skilled in the art are familiar.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The use that better implementation method described in literary composition and material only present a demonstration.
The above-mentioned feature that the present invention mentions, or the feature that embodiment mentions can combination in any.All features that patent specification discloses can with any composition forms use, each feature disclosing in description, can anyly provide the alternative characteristics of identical, impartial or similar object to replace.Therefore apart from special instruction, the feature disclosing is only the general example of equalization or similar features.
Embodiment 1: prepare Radix Astragali total saponins of the present invention
Experiment material: Milkvetch Root is purchased from Shanghai Gaoqiao Yang Hetang pharmacy; The organic reagent such as ethanol, n-butyl alcohol is analytical pure, purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Experimental technique:
Radix Astragali 5kg, is placed in CTG-10 extractor, with 70% alcohol reflux 2 times, adds 10 times of amounts the 1st time, adds 8 times of amounts the 2nd time, each 1h.Merge extractive liquid,, decompression recycling ethanol, is concentrated into 10L.With water saturated n-butyl alcohol (1: 1) extraction 3 times, merge n-butyl alcohol part, reclaim butanol extraction liquid and be concentrated into about 1L.
Extract after concentrated is slowly added on processed good D101 resin column (7.5cm * 95cm resin height 64cm), the first distilled water eluting with 4 times of column volumes, to remove carbohydrate content, continue with 30%, 70%, 90% ethanol elution of 4 times of column volumes, collect the alcohol eluen of each component, collect eluent, concentrated, be drying to obtain thick saponin.
By a small amount of dissolve with methanol of thick saponin, then add the ether of 3 times of amounts: acetone (1:1) solution, stirs, standing 10min, puts into centrifuge 5000rpm, 10min, pour out supernatant (keeping standby), precipitation is used a small amount of methanol ultrasonic dissolution again, then adds the ether of 3 times of amounts: acetone (1:1) solution, 3 times repeatedly, last precipitation is with after methanol ultrasonic dissolution, filter, reclaim solvent, drying under reduced pressure and get final product.
Embodiment 2: prepare Radix Astragali total saponins of the present invention
Experiment material: Milkvetch Root is purchased from Shanghai Gaoqiao Yang Hetang pharmacy; The organic reagent such as ethanol, n-butyl alcohol is analytical pure, purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Experimental technique:
Radix Astragali 5kg, is placed in CTG-10 extractor, with 70% alcohol reflux 2 times, adds 10 times of amounts the 1st time, adds 8 times of amounts the 2nd time, each 2h.Merge extractive liquid,, decompression recycling ethanol, is concentrated into 10L.With water saturated n-butyl alcohol (1: 1) extraction 2 times, merge n-butyl alcohol part, reclaim butanol extraction liquid and be concentrated into about 1L.
Extract after concentrated is slowly added on processed good D101 resin column (7.5cm * 95cm resin height 64cm), the first distilled water eluting with 3 times of column volumes, to remove carbohydrate content, continue with 30%, 70%, 90% ethanol elution of 3 times of column volumes, collect the alcohol eluen of each component, collect eluent, concentrated, be drying to obtain thick saponin.
By a small amount of dissolve with methanol of thick saponin, then add the ether of 4 times of amounts: acetone (1:1) solution, stirs, standing 10min, puts into centrifuge 5000rpm, 10min, pour out supernatant (keeping standby), precipitation is used a small amount of methanol ultrasonic dissolution again, then adds the ether of 3 times of amounts: acetone (1:1) solution, 4 times repeatedly, last precipitation is with after methanol ultrasonic dissolution, filter, reclaim solvent, drying under reduced pressure and get final product.
Embodiment 3: prepare Radix Astragali total saponins of the present invention
Experiment material: Milkvetch Root is purchased from Shanghai Gaoqiao Yang Hetang pharmacy; The organic reagent such as ethanol, n-butyl alcohol is analytical pure, purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Experimental technique:
Radix Astragali 5kg, is placed in CTG-10 extractor, with 70% alcohol reflux 2 times, adds 10 times of amounts the 1st time, adds 8 times of amounts the 2nd time, each 3h.Merge extractive liquid,, decompression recycling ethanol, is concentrated into 10L.With water saturated n-butyl alcohol (1: 1) extraction 3 times, merge n-butyl alcohol part, reclaim butanol extraction liquid and be concentrated into about 1L.
Extract after concentrated is slowly added on processed good D101 resin column (7.5cm * 95cm resin height 64cm), the first distilled water eluting with 4 times of column volumes, to remove carbohydrate content, continue with 30%, 70%, 90% ethanol elution of 4 times of column volumes, collect the alcohol eluen of each component, collect eluent, concentrated, be drying to obtain thick saponin.
By a small amount of dissolve with methanol of thick saponin, then add the ether of 5 times of amounts: acetone (1:1) solution, stirs, standing 10min, puts into centrifuge 5000rpm, 10min, pour out supernatant (keeping standby), precipitation is used a small amount of methanol ultrasonic dissolution again, then adds the diethyl ether solution of 3 times of amounts, 3 times repeatedly, last precipitation is with after methanol ultrasonic dissolution, filter, reclaim solvent, drying under reduced pressure and get final product.
Embodiment 4 HPLC-ELSD methods are measured the content of astragaloside in Radix Astragali total saponins of the present invention
Experiment material: acetonitrile, methanol are analytical pure, purchased from Chemical Reagent Co., Ltd., Sinopharm Group; Astragaloside: purchased from Shanghai Pu Ming Industrial Co., Ltd., lot number: AG-120802.
Experimental technique:
Chromatographic condition: Thermo Diamonsil C18 post (250 * 4.6mm, 5 μ m); Mobile phase: acetonitrile-water (36:65); Flow velocity: 1.0ml/min; Column temperature: 30 ℃; ELSD parameter: evaporating temperature: 80 ℃; Atomization temperature: 60 ℃; Outlet temperature: 50 ℃; Flow rate of carrier gas: 1.2ml/min.
After Radix Astragali total saponins sample (pressing embodiment 1～3 preparation) is dry, difference accurately weighed 5.97mg, 5.92mg, 5.89mg, add methanol 2ml and make to dissolve, with 0.45 μ m filtering with microporous membrane, stand-by.Accurately weighed astragaloside reference substance 2.53mg, adds methanol 2ml and makes to dissolve, with 0.45 μ m filtering with microporous membrane.Draw successively filtrate 5,10 μ L, respectively by above-mentioned chromatographic condition analysis, measure peak area.With the logarithm value of reference substance content (μ g), to abscissa (X), the logarithm value of reference substance peak area (A) is vertical coordinate (Y), obtains equation y=1.4053x+2.1823.
Experimental result:
As shown in Figure 1, astragaloside appearance time is 11.66min, Radix Astragali total saponins sample size is respectively 30 μ L, 20 μ L, 20 μ L, obtain peak area and be respectively 60.8, 30.4, 23.5, after substitution equation, antilogarithm converts to such an extent that Astragaloside content is respectively 0.5206 μ g, 0.3179 μ g, 0.2647 μ g, the total sample size of Radix Astragali total saponins is respectively 89.55 μ g, 59.2 μ g, 58.9 μ g, thereby show that the content of astragaloside in Radix Astragali total saponins of the present invention is respectively 0.58%, 0.54%, 0.45%, show that astragaloside is not main active in Radix Astragali total saponins of the present invention.
Embodiment 5
KM mice eight arm maze experiments
Experiment material: KM mice is provided by responsible company of Shanghai Slac Experimental Animal Co., Ltd., credit number: SYXK(Shanghai) 2009-0069; Oxiracetam injection, manufacturing enterprise: Tri-Lion Pharmaceutical Co., Ltd., Harbin, batch number: 120810B; Astragaloside, purchased from Shanghai Pu Ming Industrial Co., Ltd., lot number: AG-120802.
Experimental technique:
1) administration prospective adaptation: mice weigh after fasting 24h, eight arm ends are all placed sucrose and normal diet mixture, when each training starts, mice is placed in to labyrinth central platform, the end that freely enters eight radiation arms by it takes food, until 10min is last or fulfil looking for food of all arms ahead of schedule, finishes adaptive training one time, adapt to once every day, altogether 4d.Give during this time half diet and raise, make mice all the time in starvation.
2) administration test: 24h administration after training, blank group lumbar injection every day PBS, positive controls lumbar injection oxiracetam 800mg/kg; Low dose group lumbar injection Radix Astragali total saponins of the present invention (pressing embodiment 1 preparation) 100mg/kg; High dose group lumbar injection Radix Astragali total saponins 200mg/kg.Administration second day carries out eight arm labyrinths, four arm ends are placed sucrose and normal diet mixture 50mg therein at random, mice is placed in to labyrinth central platform, and the end that freely enters radiation arm by it takes food, until 5min end or fulfil looking for food of all arms ahead of schedule.Train continuously 5d.Mice enters the arm of food and absorbed food is a selecting properly, otherwise is wrong choice.Recording parameters is the number of times of wrong choice.Reenter and put food arm and be called working memory mistake, enter and do not put food arm and be called reference memory mistake.Omnidistance video recording.After training finishes, three weeks (administration 26d, 27d) once tests again.
Experimental result:
1. Radix Astragali total saponins significantly reduces the reference memory mistake of normal KM mice
As shown in Figure 2 A, experiment mice after the Radix Astragali total saponins of 100mg/kg is processed, 3 weeks reference memory errors number and the matched group mistake of comparing after eight arms training, number of times obviously reduces (p<0.05), and positive control drug oxiracetam group is made mistakes number of times also significantly lower than matched group (p<0.05).
2. Radix Astragali total saponins significantly improves the front positive exact figures of head mistake of the eight arm maze experiments of normal KM mice
As shown in Figure 2 B, the experiment mice after the Radix Astragali total saponins of 200mg/kg is processed, before the first mistake of eight latter 3 weeks of arm training, positive exact figures and matched group are compared, and correctly number of times obviously raises (p<0.05), even higher than positive control drug oxiracetam group.
Embodiment 6
The experiment of KM mice shuttle box
Experiment material: KM mice is provided by responsible company of Shanghai Slac Experimental Animal Co., Ltd., credit number: SYXK(Shanghai) 2009-0069; Oxiracetam injection, manufacturing enterprise: Tri-Lion Pharmaceutical Co., Ltd., Harbin, batch number: 120810B; Astragaloside, purchased from Shanghai Pu Ming Industrial Co., Ltd., lot number: AG-120802.
Experimental technique:
First mice is put into reaction chamber and adapt to 4min, to eliminate investigatory reaction, be then placed on the experimental box electric shock district of shuttling back and forth.First give conditioning stimulus 8s, after the 2s of interval, give electricity irritation 5s.If animal escapes to place of safety as escape reaction initiatively in the 10s of conditional stimulus and interval, after electric shock, just escaping to place of safety is passive escape reaction.After training for several times, mice can form initiatively escape conditional reflex gradually, forms longterm memory.After each training, after the 10s of interval, give same stimulation, repetition training 40 times.Train continuously 5d.In the time of training, give blank group lumbar injection every day PBS, positive controls lumbar injection oxiracetam 800mg/kg; Low dose group lumbar injection Radix Astragali total saponins of the present invention (pressing embodiment 2 preparations) 100mg/kg; High dose group lumbar injection Radix Astragali total saponins 200mg/kg.Administration 11d and 21d official testing, number of times initiatively escaped in record.
Experimental result:
What Radix Astragali total saponins significantly improved the experiment of normal KM mice shuttle box initiatively escapes number of times
As shown in Figure 3, after training finishes, when one week (after administration the 11st day), mice and matched group comparison after Radix Astragali total saponins is processed, initiatively escape number of times and increase significantly (p<0.05), even also higher than positive drug oxiracetam group; After administration the 21st day, 100mg/kg Radix Astragali total saponins administration group and matched group ratio, though significance level before not reaching still kept obvious advantage (p<0.07).
Conclusion:
From above-mentioned experimental result, Radix Astragali total saponins of the present invention can significantly improve the maintenance of impermanent memory, longterm memory and the memory of normal mouse, can be used as medicine or health product that normal person improves memory.
Many aspects involved in the present invention have been done as above and have been set forth.Yet, it should be understood that before not departing from spirit of the present invention and put, those skilled in the art can be equal to and change and modify it, and described change and modification fall into the coverage of the application's claims equally.
Claims (4)
1. Radix Astragali total saponins improves the medicine of memory in normal people or the application in health product in preparation.
2. application as claimed in claim 1, is characterized in that, in described Radix Astragali total saponins, the content of astragaloside is lower than 5%.
3. application as claimed in claim 2, is characterized in that, in described Radix Astragali total saponins, the content of astragaloside is lower than 1%.
4. the application as described in claim 1 or 2 or 3, is characterized in that, described Radix Astragali total saponins makes by the following method:
(a) 70% alcohol reflux of 8～10 times of amounts 2～3 times for the Radix Astragali, each 1～3h, merge extractive liquid,, decompression recycling ethanol, concentrated;
(b) by the n-butanol extraction of water saturated 1: 1 2～4 times for above-mentioned extracting solution after concentrated, combining extraction liquid, reclaims n-butyl alcohol, concentrates;
(c) by D101 type macroporous adsorbent resin on the extract after above-mentioned concentrating, first, with the distilled water eluting of 3～4 times of amounts, continue with 30% ethanol, 70% ethanol and 90% ethanol gradient elution of 3～4 times of amounts, collect eluent, concentrated, dry, obtain crude extract;
(d) by above-mentioned crude extract dissolve with methanol, then add the ether acetone soln of the 1:1 of 3～5 times of amounts, stir, standing, centrifugal, precipitation is used dissolve with methanol again, and 3～5 times repeatedly, filter, reclaim methanol, concentrated, dry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310250408.XA CN103285071B (en) | 2013-06-21 | 2013-06-21 | Medical use of milkvetch root total saponin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310250408.XA CN103285071B (en) | 2013-06-21 | 2013-06-21 | Medical use of milkvetch root total saponin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103285071A CN103285071A (en) | 2013-09-11 |
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| CN108929357B (en) * | 2018-08-14 | 2021-05-07 | 安徽弘腾药业有限公司 | Method for rapidly extracting astragaloside from astragalus |
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| 李维祖.黄芪总苷及黄芪甲苷对糖皮质激素诱导老前期小鼠记忆障碍的保护作用及其机制.《安徽医药》.2008,第12卷(第7期),592-594. * |
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