CN105687564A - Traditional Chinese medicine composition for improving sleep and memory and preparation method of traditional Chinese medicine composition for improving sleep and memory - Google Patents
Traditional Chinese medicine composition for improving sleep and memory and preparation method of traditional Chinese medicine composition for improving sleep and memory Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine composition for improving sleep and memory and a preparation method of the traditional Chinese medicine composition for improving the sleep and the memory. The preparation method of the traditional Chinese medicine composition mainly includes extracting Radix Codonopsis, Poria Cocos and Radix Polygalae by ethanol, conducting ethanol precipitation and edulcoration, passing a macroporous resin column for purification, drying and mixing a dried product with superfine Rhizoma Acori Tatarinowii powder so as to obtain the traditional Chinese medicine composition. Results of animal pharmacological experiments and clinical tests show that the traditional Chinese medicine composition is superior to the prior art in sleep and memory improvement.
Description
Technical field
The present invention relates to pharmaceutical sanitary field, it is specifically related to a kind of Chinese medicine composition improving sleep and memory and preparation method thereof。
Background technology
The problem of insomnia, of common occurrence in daily life at present。The life of people is caused serious impact by long-term insomnia, it is possible to the daily vital function of interference people, bring out multiple disease, concurrent various Negative Emotionals。
Insomnia is referred to as " being insomnia " in the traditional Chinese medical science, is insomnia and is called " must not crouch ", " unable to close the eyes " at Huangdi's Internal Classics, and main manifestations is the deficiency of the length of one's sleep, the degree of depth。The pathogen and pathology of tcm of insomnia is mainly diseases caused by exogenous pathogenic factor six climate exopathogens, incoordination between the spleen and stomach, negative and positive of qi and blood is unbalance, internal damage by the excessive seven emotions etc., and Syndrome Types of TCM has type of deficiency of both the heart and spleen, type of hyperactivity of fire caused by deficiency of YIN, type of deficiency of heart-QI and gallbladder-QI, type of phlegm-heat attacking internally, fire derived from stagnation of liver-QI type。
The high incidence of insomnia is well proved, in crowd, the insomnia outbreak that the adult having 1/3 is once above in a year is estimated in investigation。One of investigation that the U.S. is best shows, the surveyee of 15% in the previous year in have a serious insomnia problem, insomnia increases the weight of with advancing age, and in intermediate ages sample cluster, chronic insomnia rate is about 10%。It is 25% even higher for having the insomnia sickness rate of clinical meaning in the adult that the age is bigger。Insomnia is so common to such an extent as to become one of characteristic negativity change in aging course in old people。
Hypomnesis, the traditional Chinese medical science claims forgetful, forgetful, forgetfulness, and it is the aging of brain physiological and pathological seaility。Forgetful etiology and pathogenesis is all had discussion by ancient Chinese medicine doctor。"Nei Jing" is referred to as " forgetful "。(the big puzzled opinion of Ling Shu Miraculous Pivot or Divine Axis " propose: " QI of the upper portion of the body being insufficient, the therapeutic method to keep the adverse QI flowing downwards is had a surplus, the intestines and stomach is real and cardiopulmonary are empty, void then seek defend stay under, of a specified duration not with time on, therefore forgetful also。" think forgetful be because of seek the stagnation of QI defended in lower and can not on moisten caused by。The Northern Song Dynasty " Shengji Zonglu " is said: " forgetful disease, this is in having a guilty conscience, and vim and vigour decline few, spiritual decrepit and muddleheaded, therefore will turmoil and poor memory are also。" Southern Song Dynasty sternly with and recipes for Saving Lives emphasize that primary disease and heart spleen are closely related: " lid spleen idea and think of, the heart is main think of also, excessive thinking, and Yishe (US 49) is unclear, god's official's not duty, makes people forgetful。" unit's Zhu Zhenheng " red the small stream method for the treatment of heart want " thinks that primary disease is except main heart spleen, " the person that also has mental confusion due to phlegm。" bright Li Zhongzi " Required Readings for Medical Professionals " thinks that this cause of disease " disarmony between the heart and kidney " causes。Clear Lin Peiqin " Lei Zheng Zhi Cai " plays saying of Li Shizhen (1518-1593 A.D.) " the brain being the house of mentality ", it is recognised that " real memory institute is with also for the brain being the house of mentality, the sea of marrow " is also attributed to caused by brains deficiency forgetful。Clear Tang ancestor sea " treatise on blood trouble " says: " all hearts have blood stasis, also make forgetful。" in a word, a forgetful card is many by deficiency of both the heart and spleen, disarmony between the heart and kidney, essential resistance of turbid phlegm, the factor such as stop in blood stasis finally have impact on the function of brain (brain) and causes。
The 3rd disclosed mind tranquilizing pills of " the Sanitation Ministry medicine standard " Traditional Chinese medicine historical preparation discloses following information:
1, composition: Radix Codonopsis 300g, Poria 300g, Radix Polygalae 200g, Rhizoma Acori Graminei 200g。
2, method for making: above four tastes, is ground into fine powder, sieves, mixing。It is appropriate with water that every 100g powder adds refined honey 35～45g, general ball, the another Cinnabaris 2g after flying that fetches water, coating, and cold drying to obtain final product。
3, function with cure mainly: benefiting vital QI for tranquillizing, restoring normal coordination between the heart and kidney, improving eyesight。Not peaceful for mind, palpitate with fear forgetful, insomnia, asthenia, visual deterioration。
In described ministry standard, the method for making of mind tranquilizing pills is that full medicated powder is used as medicine, and there is effective ingredient dissolution few, the problem that bioavailability is low。Accordingly, it is desirable to provide a kind of preparation method that can improve medicine drug effect。
Summary of the invention
The preparation method that it is an object of the invention to provide the Chinese medicine composition improving sleep and memory of a kind of determined curative effect。
A kind of Chinese medicine composition improving sleep and memory provided by the invention, the preparation method of its active component comprises the following steps:
1) medical material of following weight portion is taken: Radix Codonopsis 3 parts, 3 parts of Poria, Radix Polygalae 2 parts, Rhizoma Acori Graminei 2 parts；
2) Radix Codonopsis, Poria, Radix Polygalae extracting method: after adding the ethanol extraction that percent by volume is 20%-40%, adding percent by volume is that 85%-95% ethanol carries out precipitate with ethanol, and supernatant crosses macroporous resin column, eluting, collects eluent, and concentration is dry, makes dried cream powder；
3) Rhizoma Acori Graminei is crushed into impalpable powder；
4) by step 2), 3) gained dried cream powder and impalpable powder mixing, to obtain final product。
Described above prepare a kind of improve sleep and memory Chinese medicine composition step in:
Preferably, step 2) described in step 2) described in macroporous resin be: HPD-100 macroporous resin。
Preferably, step 3) described in breaking method be: superfine comminution at low temperature。
Preferably, step 2) described in Radix Codonopsis, Poria, the extracting method of Radix Polygalae is: by Radix Codonopsis, Poria and Radix Polygalae powder are broken into coarse powder, adding concentration of volume percent is 20-40% soak with ethanol 24h, reflux, extract, 2-3 time, each ethanol consumption is 8-12 times of medical material gross weight, each extraction time is 1.5-3h, united extraction liquid, being evaporated to crude drug concentration is 0.3-0.6g/ml, the ethanol adding 3-5 times of 85-95% carries out precipitate with ethanol, the precipitate with ethanol time is 12-24 hour, filter, centrifugal, supernatant concentration to crude drug concentration is 0.3-0.6g/ml, upper HPD-100 macroporous resin column carries out secondarily purified, loading volume is 3-4 times of resin column volume, loading flow velocity is 2-4BV/h, carbohydrate content is eluted with 1-2BV/h with the water of 5-7 times of column volume, continue and with 1-2BV/h, saponin component is eluted with 5-7 times of column volume 30-60% ethanol, merge eluent, dry, obtain。
Described step 2) described in extracting method much further preferably from: by Radix Codonopsis, Poria and Radix Polygalae powder are broken into coarse powder, adding concentration is 30-40% soak with ethanol 24h, reflux, extract, 2-3 time, each ethanol consumption is 10-12 times of medical material gross weight, each extraction time is 2-3h, united extraction liquid, being evaporated to concentration is 0.5-0.6g/ml, the ethanol adding 4-5 times of 85-95% carries out precipitate with ethanol remove impurity, the precipitate with ethanol time is 12-24 hour, filter, centrifugal (2000r/min-4000r/min) 10-20min, filtration discards precipitation, being concentrated into crude drug concentration is 0.3-0.6g/ml, take macroporous resin column on this medicinal liquid and carry out secondarily purified, loading volume is 3-4 times of resin column volume, loading flow velocity is 3-4BV/h, with the water of 6-7 times of column volume, carbohydrate content is eluted, continue and with 5-7 times of column volume 30-60% ethanol, saponin component is eluted, merge eluent, dry to obtain mixed extract。
Present invention also offers the preparation containing above-mentioned Chinese medicine composition, said preparation is separately made by Chinese medicine composition or is made up of Chinese medicine composition and pharmaceutically acceptable carrier。
Described preparation is solid preparation, such as tablet, capsule or granule etc.；
Described pharmaceutically acceptable carrier is one or more in lactose, starch, dextrin, mannitol, sucrose, hydroxypropyl methylcellulose, cross-linking sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, microcrystalline Cellulose, micropowder silica gel, pregelatinized Starch, sorbitol, magnesium stearate, Pulvis Talci, hydroxypropyl methylcellulose, low-substituted hydroxypropyl cellulose, sodium benzoate, potassium sorbate, sucrose or aspartame, monosodium glutamate, Mel, orange flavor。
The preparation method of Chinese medicinal composition preparation of the present invention, the method can adopt the preparation of conventional formulation technique for various common dosage forms。
Such as:
Method 1: Chinese medicine composition pill of the present invention
Taking the present invention and prepare gained active ingredient of Chinese herbs, add the conventional adjuvant of pill, production method is prepared into the pill of Chinese medicine composition of the present invention routinely。
Method 2: Chinese medicinal composition capsules agent of the present invention
Taking the present invention and prepare gained active ingredient of Chinese herbs, add the conventional adjuvant of capsule, production method is prepared into the capsule of Chinese medicine composition of the present invention routinely。
Method 3: Chinese medicine composition tablet of the present invention
Taking the present invention and prepare gained active ingredient of Chinese herbs, add the conventional adjuvant of tablet, production method is prepared into the tablet of Chinese medicine composition of the present invention routinely。
Method 4: Chinese medicinal composition granules of the present invention
Taking the present invention and prepare gained active ingredient of Chinese herbs, add the conventional adjuvant of granule, production method is prepared into the granule of Chinese medicine composition of the present invention routinely。
Method 5: Chinese medicine composition syrup of the present invention
Taking the present invention and prepare gained active ingredient of Chinese herbs, add the conventional adjuvant of syrup, production method is prepared into the syrup of Chinese medicine composition of the present invention routinely。
Present invention also offers the application of Chinese medicine composition improving sleep and memory and preparation method thereof in preparation of above-mentioned Chinese medicine composition or preparation。
Chinese medicine composition effective ingredient disclosed in this invention is to be prepared from by raw materials such as Radix Codonopsis, Poria, Radix Polygalae, Rhizoma Acori Graminei。Inventor finds in preparation research process for many years, identical drug dose, the active component of different extraction process gained, also can there is bigger difference in its pharmacological action, for this, the Chinese medicine composition of the present invention has been carried out Study on extraction in conjunction with pharmacological evaluation by applicant, sum up the process for extracting of the present invention, and experiments verify that such improvement brings drug effect potentiation: work as Radix Codonopsis, Poria, Polygala tenuifolia alcohol extraction purified and Grassleaf Sweetflag Rhizome mixing gained Chinese medicine composition, effect for improving sleep and memory is far superior to the existing technology of preparing of identical prescription, there is significance progress。
Owing to every taste Chinese crude drug all contains many active component, at least not yet there is any a herb purely and simply to be analyzed and know all the components, different Chinese medicine combines when extracting with different extracting modes, due to influencing each other and acting between solvent, temperature and heterogeneity, extremely complex chemical reaction can be produced。Therefore, above-mentioned effective ingredient is a part for Chinese medicine extract active component, and the drug action impact for overall medicine is still not clear。We contrast different extracting mode quality in improving the purposes of sleep and memory only by effect experiment at present, and the method that there is no changes from the change indicator of part effective ingredient to the curative effect judging compound Chinese patent medicine。
By " extending the pentobarbital sodium experiment length of one's sleep ", the different extracting mode curative effect in improving sleep of " pentobarbital sodium sub-threshold lull dosage experiment " contrast；We pass through step down test, Morris water maze test, the different extracting mode curative effect in improving memory of contrast。
The extracting method of mind tranquilizing pills has been carried out systematic research experiment with the excellent effect of medicine by inventor, by extracting with diverse ways, finally obtained the extractive technique scheme of the present invention, made the medical material of same recipe, given play to therapeutic purposes more clearly, the effect of more remarkable treatment effect。
A kind of Chinese medicine composition improving sleep and memory provided by the invention has the advantage that
1, Chinese medicine composition provided by the invention can be obviously prolonged mouse sleep time, significant difference (P < 0.01) compared with blank group, and compared with comparative example group, all there is significant difference (P < 0.05, P < 0.01)；Compare with former technique group, all there is significant difference (P < 0.05, P < 0.01)。
2, Chinese medicine composition provided by the invention can substantially increase mice sleep rate in 30min, significant difference (P < 0.05, P < 0.01) compared with blank group；And compared with comparative example group, all there is significant difference (P < 0.05, P < 0.01)。
The above results shows, Chinese medicine composition provided by the invention improves the effect of sleep and is better than prior art。
3, Chinese medicine composition provided by the invention has the effect of improvement, consolidating memory, compares with model group, and mice step down test errors number significantly reduces (P < 0.05, P < 0.01)。
4, scopolamine is caused the cognitive disorder of mice by Chinese medicine composition provided by the invention improvement result, compare with model control group, the incubation period relatively model group of appearing on the stage substantially shortens (P < 0.05 or P < 0.01), and errors number significantly reduces (P < 0.05 or P < 0.01)；Comparing with comparative example group, the incubation period relatively model group of appearing on the stage substantially shortens (P < 0.05 or P < 0.01)；Comparing with former technique group, the incubation period relatively model group of appearing on the stage substantially shortens (P < 0.05 or P < 0.01)。
5, this Chinese prescription is carried out extraction purification by the present invention further so that dosage is less, and effect is more excellent。
Detailed description of the invention
Further illustrate the present invention by the examples below。It should be understood that embodiments of the invention are an illustration for the present invention rather than limitation of the present invention。The simple modifications that the present invention is carried out by the essence according to the present invention broadly falls into the scope of protection of present invention。Except as otherwise noted, the percent of the amount of alcohol in the present invention is percentage by volume。
Embodiment 1:
Take Radix Codonopsis 3kg respectively, Poria 3kg and Radix Polygalae 2kg is ground into coarse powder, adding concentration is 30% soak with ethanol 24h, reflux, extract, 2-3 time, each ethanol consumption is 10 times of medical material gross weight, each extraction time is 2h, united extraction liquid, being evaporated to raw concentration is 0.5g/ml, the ethanol adding 4 times of 85-95% carries out precipitate with ethanol remove impurity, the precipitate with ethanol time is 12h, filter, centrifugal (3000r/min) 15min, filtration discards precipitation, being concentrated into crude drug concentration is 0.5g/ml, take this medicinal liquid to cross HPD-100 macroporous resin column and carry out secondarily purified, loading volume is 4 times of resin column volume, loading flow velocity is 3BV/h, with the water 1BV/h of 6 times of column volumes, carbohydrate content is eluted, continue and with 1BV/h, saponin component is eluted with 6 times of column volume 40% ethanol, merge eluent, dry to obtain mixed extract, again this mixed extract mixed homogeneously with the Rhizoma Acori Graminei crushed and get final product。
Embodiment 2:
Take Radix Codonopsis 3kg respectively, Poria 3kg and Radix Polygalae 2kg is ground into coarse powder, adding concentration is 40% soak with ethanol 24h, reflux, extract, 2 times, each ethanol consumption is 12 times of medical material gross weight, each extraction time is 2h, united extraction liquid, being evaporated to raw concentration is 0.5g/ml, the ethanol adding 5 times 85% carries out precipitate with ethanol remove impurity, the precipitate with ethanol time is 12h, filter, centrifugal (2000r/min) 20min, filtration discards precipitation, being concentrated into crude drug concentration is 0.5g/ml, take this medicinal liquid to cross HPD-100 macroporous resin column and carry out secondarily purified, loading volume is 4 times of resin column volume, loading flow velocity is 4BV/h, carbohydrate content is eluted with 2BV/h with the water of 6 times of column volumes, continue and with 2BV/h, saponin component is eluted with 6 times of column volume 40% ethanol, merge eluent, dry to obtain mixed extract, again this mixed extract mixed homogeneously with the Rhizoma Acori Graminei crushed and get final product。
Embodiment 3:
Take Radix Codonopsis 3kg respectively, Poria 3kg and Radix Polygalae 2kg is ground into coarse powder, adding concentration is 40% soak with ethanol 24h, reflux, extract, 3 times, each ethanol consumption is 10 times of medical material gross weight, each extraction time is 3h, united extraction liquid, being evaporated to raw concentration is 0.5g/ml, the ethanol adding 4 times 85% carries out precipitate with ethanol remove impurity, the precipitate with ethanol time is 18h, filter, centrifugal (3000r/min) 15min, filtration discards precipitation, being concentrated into crude drug concentration is 0.5g/ml, take this medicinal liquid to cross HPD-100 macroporous resin column and carry out secondarily purified, loading volume is 4 times of resin column volume, loading flow velocity is 3BV/h, carbohydrate content is eluted with 1BV/h with the water of 5 times of column volumes, continue and with 1BV/h, saponin component is eluted with 5 times of column volume 60% ethanol, merge eluent, dry to obtain mixed extract, again this mixed extract mixed homogeneously with the Rhizoma Acori Graminei crushed and get final product。
Embodiment 4:
Take Radix Codonopsis 3kg respectively, Poria 3kg and Radix Polygalae 2kg is ground into coarse powder, adding concentration is 20% soak with ethanol 24h, reflux, extract, 3 times, each ethanol consumption is 12 times of medical material gross weight, each extraction time is 2h, united extraction liquid, being evaporated to raw concentration is 0.5g/ml, the ethanol adding 3 times 95% carries out precipitate with ethanol remove impurity, the precipitate with ethanol time is 24h, filter, centrifugal (4000r/min) 20min, filtration discards precipitation, being concentrated into crude drug concentration is 0.5g/ml, take this medicinal liquid to cross HPD-100 macroporous resin column and carry out secondarily purified, loading volume is 4 times of resin column volume, loading flow velocity is 3BV/h, carbohydrate content is eluted with 2BV/h with the water of 5 times of column volumes, continue and with 2BV/h, saponin component is eluted with 5 times of column volume 50% ethanol, merge eluent, dry to obtain mixed extract, again this mixed extract mixed homogeneously with the Rhizoma Acori Graminei crushed and get final product。
Embodiment 5: pill
1, composition: the thick paste of embodiment 1 gained, is dried to dried cream powder, obtains 148g, add refined honey 60g, Cinnabaris 2g。
2, preparation method: the method with reference to conventional Chinese medicine preparation makes pill。
Embodiment 6: capsule
1, composition: the thick paste of embodiment 2 gained, is dried to dried cream powder, obtains 152g, adds mannitol 32g, magnesium stearate 2.8g, Pulvis Talci 2.8g。
2, preparation method: the method with reference to conventional Chinese medicine preparation makes capsule。
Embodiment 7: tablet
1, composition: the thick paste of embodiment 3 gained, is dried to dried cream powder, obtains 158g, adds microcrystalline Cellulose 50g, cross-linking sodium carboxymethyl cellulose 4.0g, micropowder silica gel 2.5g。
2, preparation method: the method with reference to conventional Chinese medicine preparation makes tablet。
Embodiment 8: granule
1, composition: the thick paste of embodiment 4 gained, is dried to dried cream powder, obtains 154g, adds dextrin 650g, pregelatinized Starch 75g。
2, preparation method: the method with reference to conventional Chinese medicine preparation makes granule。
Former technique:
Reference the 3rd disclosed mind tranquilizing pills preparation method of " the Sanitation Ministry medicine standard " Traditional Chinese medicine historical preparation, specific as follows:
Weigh Radix Codonopsis 300g, Poria 300g, Radix Polygalae 200g, Rhizoma Acori Graminei 200g；
Above four tastes, are ground into fine powder, sieve, mixing, to obtain final product。
Experimental example: the comparison of Chinese medicine composition extracting method
1, sample packet: embodiment 1-4, former process component do not make dried cream powder。
2, contrast project
Total polysaccharides content: with colorimetric method for determining total polysaccharides content；
Total saponin content: vanilla root rot development process；
The rate of extract: hypobaric drying method。
3, experimental result is in Table 1
The different percolation method extraction effect to medical material of table 1
| Group | Total saponins (mg g-1) | Total polysaccharides (mg g-1) | The rate of extract (%) |
| Embodiment 1 | 9.3±1.16 | 146.8±1.23 | 14.8±1.37 |
| Embodiment 2 | 10.8±1.04 | 151.7±1.15 | 15.2±1.30 |
| Embodiment 3 | 11.2±1.11 | 163.4±1.12 | 15.8±1.33 |
| Embodiment 4 | 12.7±1.03 | 170.5±1.20 | 15.4±1.36 |
| Former technique group | 8.9±1.22 | 135.8±1.31 | - |
Table 1 result shows: the total saponins of embodiment of the present invention 1-4 and the content of total polysaccharides are superior to former technique。Experimental result points out the technology of the present invention to extract gained Chinese medicine composition total saponins, total polysaccharides compared with prior art, extracts more complete。
Effect experiment
One, the pentobarbital sodium experiment length of one's sleep is extended
1, experiment material and method
1.1 experiment materials
Healthy SPF level Kunming mouse, male, body weight 20 ± 2g, buys from Guangxi Medical University's Experimental Animal Center (production licence number: SCXK osmanthus 2009-0002)。
Embodiment of the present invention 1-4；Former technique example；Comparative example (melatonin sheet)；Pentobarbital sodium (sigma)。1.2 methods
Mice adaptability feeds 3d, takes 84 mices, is randomly divided into blank group, comparative example group, former technique group, embodiment 1-4 group, often 12 mices of group。Carrying out gavage to sample by following dosage: former technique group is 1000mg/kg/d, embodiment group dosage is 100mg/kg/d, and contrast groups dosage is 0.5mg/kg/d, and blank group gives isometric(al) distilled water, 0.2mL/10g, 1 time/d, continuous 30d。
After last administration 15min, each group mouse peritoneal injection 50mg/kg pentobarbital sodium, injection volume is 0.2mL/20g, with righting reflex loss (when mice is placed in supine position, in 60s can not the person of righting, namely think righting reflex loss) for index, observe the length of one's sleep。Variance analysis is adopted to add up。
2, result is in Table 2
Table 2 on extend pentobarbital sodium length of one's sleep impact (N=12)
Note: compare with blank group, * P < 0.05, * * P < 0.01；Compare with comparative example group,#P < 0.05,##P < 0.01；Compare with former technique group,▲P < 0.05,▲▲P < 0.01。
Table 2 result shows, compares with blank group, and comparative example group, former technique group and embodiment 1-4 group all can be obviously prolonged mouse sleep time (P < 0.01)；Comparing with comparative example group, embodiment 1-4 group all can be obviously prolonged mouse sleep time (P < 0.05 or P < 0.01)；Comparing with former technique group, embodiment 1-4 group all can be obviously prolonged mouse sleep time (P < 0.05 or P < 0.01)。
More than test result indicate that, the embodiment of the present invention enhances the health care improving sleep after changing original technique。
Two, pentobarbital sodium sub-threshold lull dosage experiment
1.1 experiment materials
Healthy SPF level Kunming mouse, male, body weight 20 ± 2g, buys from Guangxi Medical University's Experimental Animal Center (production licence number: SCXK osmanthus 2009-0002)。
Embodiment of the present invention 1-4；Former technique example；Comparative example (melatonin sheet)；Pentobarbital sodium (sigma)。
1.2 methods
Take 140 mices, it is randomly divided into blank group, comparative example group, former technique group, embodiment 1-4 group, often group 20, carrying out gavage to sample by following dosage: embodiment group dosage is 100mg/kg/d, contrast groups dosage is 0.5mg/kg/d, former technique group dosage 1000mg/kg/d, blank group gives isometric(al) distilled water, 0.2mL/10g, 1 time/d, continuous 30d。
After last administration 15min, each group mouse peritoneal injection maximum subthreshold hypnotic dosage of 30mg/kg pentobarbital sodium, injection volume is 0.2mL/20g, observes (the above person of righting reflex loss 1min) number of mice of falling asleep in 30min。
2, result is in Table 3
Table 3 on the impact of pentobarbital sodium sub-threshold lull dosage experiment mice (N=20)
Note: compare with blank group, * P < 0.05, * * P < 0.01；Compare with comparative example group,#P < 0.05,##P < 0.01；Compare with former technique group,▲P < 0.05,▲▲P < 0.01。
Table 3 result shows, compares with blank group, and comparative example group, former technique group and embodiment 1-4 group all can substantially increase mice sleep rate (P < 0.05 or P < 0.01) in 30min；Comparing with comparative example group, embodiment 1-4 group all can substantially increase mice sleep rate (P < 0.05 or P < 0.01) in 30min；Comparing with former technique group, embodiment 2-4 group all can substantially increase mice sleep rate (P < 0.05 or P < 0.01) in 30min。
More than test result indicate that, the embodiment of the present invention enhances the health care improving sleep after changing original technique。
Three, step down test
1, experiment material and method
1.1 experiment materials
Healthy SPF level Kunming mouse, male and female dual-purpose, body weight 20 ± 2g, buys from Guangxi Medical University's Experimental Animal Center (production licence number: SCXK osmanthus 2009-0002)。
Embodiment of the present invention 1-4, former technique example, comparative example (Piracetam Tablet), scopolamine
1.2 methods
Mice 80,2 monthly age adult mices, it is randomly divided into 8 groups, often group 10 animals, respectively blank group, model control group, comparative example group, former technique group and embodiment 1-4 group。
Each group of embodiment 1-4 gives test sample 100mg/kg/d, every day gavage 1 time；Blank group and model control group give 0.9% sodium chloride solution, and former technique group dosage is 1000mg/kg/d, comparative example group dosage 1200mg/kg/d, continuous 30d。Animal is weighed weekly twice, to adjust given the test agent dosage。Training in 30th day。At first 10 minutes lumbar injection scopolamine normal saline solutions of training, blank group injection normal saline。Each group of the every batch of experiment has a mice to test sample, operation repetitive, by that analogy。Being respectively put in 4 grid of diving tower instrument by first 4 mices during training, first adapt to environment 3 minutes, be then electrified to, after mice is shocked by electricity, majority jumps onto diving tower, escapes electric shock。Contact copper grid for getting an electric shock with mice biped when jumping off simultaneously, be considered as wrong reaction, train 5 minutes, and record electric shock number of times in 5 minutes, retest after 24 hours, with incubation period and electric shock number of times for observation index record test result。
2, experimental result is in Table 4
Table 4 each Mus mice step down test result (N=10)
Note: compare with blank group, * P < 0.05, * * P < 0.01；Compare with model control group,#P < 0.05,##P < 0.01；Compare with comparative example group,▲P < 0.05,▲▲P < 0.01；Compare with former technique group,&P < 0.05,&&P < 0.01。
Table 4 result shows, compares with blank group, and model control group substantially shortens incubation period, electric shock number of times substantially increases (P < 0.01)；Comparing with model control group, administration group is obviously prolonged incubation period, the number of times that gets an electric shock significantly reduces (P < 0.05 or P < 0.01)；Comparing with comparative example group, embodiment is obviously prolonged 3,4 groups of incubation periods, the number of times that gets an electric shock significantly reduces (P < 0.05 or P < 0.01)；Comparing with former technique group, embodiment is obviously prolonged 3,4 groups of incubation periods, the number of times that gets an electric shock significantly reduces (P < 0.05 or P < 0.01)。
Test result indicate that, embodiment 1-4 group reduces (P < 0.05 or P < 0.01) relative to model control group prolongation of latency and errors number, embodiment 3,4 groups reduces (P < 0.05 or P < 0.01) relative to former technique group group prolongation of latency and errors number, and new technology can strengthen the effect of mice improvement, consolidating memory。
Four, Morris water maze test
1, experiment material and method
1.1 experiment materials
Healthy SPF level Kunming mouse, male and female dual-purpose, body weight 20 ± 2g, buys from Guangxi Medical University's Experimental Animal Center (production licence number: SCXK osmanthus 2009-0002)。
Embodiment of the present invention 1-4, former technique example, comparative example (Piracetam Tablet), scopolamine
1.2 methods
Mice 80,2 monthly age adult mices, be randomly divided into 8 groups, often 10 animals of group, respectively blank group, model control group, comparative example group, former technique group and embodiment 1-4 group。
Each group of embodiment 1-4 gives test sample 100mg/kg/d, every day gavage 1 time；Blank group and model control group give 0.9% sodium chloride solution, and former technique group dosage is 1000mg/kg/d, and comparative example group dosage is 1200mg/kg/d, continuous 30d。Animal is weighed weekly twice, to adjust given the test agent dosage。Within 30th day, start training。At first 10 minutes lumbar injection scopolamine normal saline solutions of training, blank group injection normal saline。Experiment allows mice free swimming 2min to adapt to surrounding the previous day, and from daystart, the every batch of experiment is respectively arranged with 1 mice to test sample, operation repetitive, by that analogy。Training every day 4 times, randomly choose a place of entry every time, put in water by mice towards pool wall, the labyrinth video camera with display system disposed above, computer is from motion tracking timing and records swimming track。4 times training mice enters water from four different place of entry respectively, if mice does not find platform in 120s, need to be caused platform。At this moment it is designated as 120s incubation period, every time training interval 60s, using the average times of 4 times as the training achievement of every day, training 5 days continuously。Orientation navigation is tested, and test lasts 5d, trains 4 every day。4 times incubation period achievement meansigma methods enter last statistics as the final result on the same day。Space exploration test (Spatialprobetest), 5th afternoon carries out space exploration test, removing platform, put in water at Northwest Quadrant pool wall midpoint towards pool wall by test mice, in record 60s, the number of times of mice spanning platform is as the Memory result of mice。
2, experimental result is in Table 5
Table 5 respectively organize water maze test in mice result (N=10)
Note: compare with blank group, * P < 0.05, * * P < 0.01；Compare with model control group,#P < 0.05,##P < 0.01；Compare with comparative example group,▲P < 0.05,▲▲P < 0.01；Compare with former technique group,&P < 0.05,&&P < 0.01。
Table 5 experimental result shows, compares with blank group, and the incubation period of appearing on the stage of model control group mice is obviously prolonged, and the number of times (i.e. errors number) of spanning platform substantially increases (P < 0.01)；Compare with model control group, the incubation period relatively model group of appearing on the stage each administration treated animal substantially shortens (P < 0.05 or P < 0.01), and errors number significantly reduces (P < 0.05 or P < 0.01)；Compare with comparative example group, the incubation period relatively model group of appearing on the stage embodiment 1-4 treated animal substantially shortens (P < 0.05 or P < 0.01), and 4 groups of errors number of embodiment significantly reduce (P < 0.05)；Compare with former technique group, the incubation period relatively model group of appearing on the stage embodiment 1-4 treated animal substantially shortens (P < 0.05 or P < 0.01), and 4 groups of errors number of embodiment significantly reduce (P < 0.05)。
It is shown that the cognitive disorder that scopolamine is caused mice by embodiment of the present invention 1-4 has improvement result, embodiment 4 groups improves memory effect optimum。New technology can strengthen the improvement result of mice cognitive disorder。
Although, above use generality explanation, detailed description of the invention and test, the present invention is described in detail, but on basis of the present invention, it is possible to it is made some modifications or improvements, and this will be apparent to those skilled in the art。Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention。
Claims (5)
1. the Chinese medicine composition improving sleep and memory, it is characterised in that the preparation method of its active component comprises the following steps:
1) medical material of following weight portion is taken: Radix Codonopsis 3 parts, 3 parts of Poria, Radix Polygalae 2 parts, Rhizoma Acori Graminei 2 parts；
2) Radix Codonopsis, Poria, Radix Polygalae extracting method: after adding the ethanol extraction that percent by volume is 20%-40%, adding percent by volume is that 85%-95% ethanol carries out precipitate with ethanol, and supernatant crosses macroporous resin column, eluting, collects eluent, and concentration is dry, makes dried cream powder；
3) Rhizoma Acori Graminei is crushed into impalpable powder；
4) by step 2), 3) gained dried cream powder and impalpable powder mixing, to obtain final product。
2. according to claim 1 improve sleep and the Chinese medicine composition of memory, it is characterised in that step 2) described in macroporous resin be: HPD-100 macroporous resin。
3. the Chinese medicine composition improving sleep and memory according to claim 1, it is characterised in that the breaking method described in step 3) is: superfine comminution at low temperature。
4. the Chinese medicine composition improving sleep and memory according to claim 1, it is characterized in that, step 2) described in Radix Codonopsis, Poria, the extracting method of Radix Polygalae is: by Radix Codonopsis, Poria and Radix Polygalae powder are broken into coarse powder, adding concentration of volume percent is 20-40% soak with ethanol 24h, reflux, extract, 2-3 time, each ethanol consumption is 8-12 times of medical material gross weight, each extraction time is 1.5-3h, united extraction liquid, being evaporated to crude drug concentration is 0.3-0.6g/ml, the ethanol adding 3-5 times of 85-95% carries out precipitate with ethanol, the precipitate with ethanol time is 12-24 hour, filter, centrifugal, supernatant concentration to crude drug concentration is 0.3-0.6g/ml, upper HPD-100 macroporous resin column carries out secondarily purified, loading volume is 3-4 times of resin column volume, loading flow velocity is 2-4BV/h, carbohydrate content is eluted with 1-2BV/h with the water of 5-7 times of column volume, continue and with 1-2BV/h, saponin component is eluted with 5-7 times of column volume 30-60% ethanol, merge eluent, dry, obtain。
5. improving a Chinese medicine preparation for sleep and memory, the Chinese medicine composition improving sleep and memory described in claim 1 prepares with acceptable carrier in pharmaceuticals industry and forms。
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