CN105055782A - Traditional Chinese medicine combination capable of improving memory and preparation method thereof - Google Patents
Traditional Chinese medicine combination capable of improving memory and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine combination capable of improving memory, which belongs to the technical filed of Chinese medicine. The traditional Chinese medicine combination capable of improving memory which adopts radix rehmanniae preparata as a monarch drug according to a specific radio can obviously improve scopolamine acquired memory disorder model memory, and improves memory effect which is better than existing drugs in a step-down test, a step-through test, a water maze test and a through pass test. The traditional Chinese medicine combination capable of improving memory can be prepared to be a commonly used oral dosage form, such as granules, tablets, capsule and oral liquid and the like. The traditional Chinese medicine combination capable of improving memory does not have acute toxicity, genetic toxicity and chronic toxicity, and is suitable to improve memory and prevent memory decline and memory disorders.
Description
Technical field
The application relates to the field of Chinese medicines and field of food health care, is specifically related to Chinese medicine composition of a kind of Improving memory and preparation method thereof.
Background technology
Memory be human brain to experience cross things memorize, maintenance, reproduce or re-recognize, it is the basis of carrying out the senior mental activity such as thinking, the imagination.Human memory ties with cerebral hippocampus, the chemical composition change of brain inside is relevant.Cause the main cause of hypomnesis to have the unhealthy emotions such as depression, anxiety, indignation, insomnia, disease, the age, brain overwork, excessive mental pressure, smoke, the bad hobby such as to drink.After entering person in middle and old age, with advancing age, cerebral function fails, and hypomnesis is one of middle-aged and elderly people body function debilitating symptoms.
The too early decline of memory belongs to the sick category such as the traditional Chinese medical science " forgetful ", " forgetfulness ", " poor memory ", " forgetting well ", and the traditional Chinese medical science thinks forgetful and forgetfulness, is due to excessive thinking, nervous exhaustion and causing, controls and should nourish heart kidney.Because " in the heart, the essence of the heart depends on kidney to the refreshing residence of people ", and the brain being the house of mentality, the sea of marrow, real memory institute with also.The classic of TCM has cloud: control the person forgotten and must hand over its heart kidney, lead under making the god of the heart in kidney, the elite of kidney rises in brain.Essence can be angry, and gas can give birth to god, and god determines gas clearly from the fresh mistake forgotten.Current kinds of traditional Chinese medicines has been proved the effect having and remember and improve, as the Chinese medicine of cerebral blood flow can be increased: Radix Salviae Miltiorrhizae, Radix Notoginseng, Rhizoma Chuanxiong, Radix Puerariae, Flos Carthami, Folium Ginkgo, Tong Fructus Tribuli, Radix Stephaniae Tetrandrae, Caulis Piperis Kadsurae, Radix Ilicis Pubescentis, Cortex Ilicis Rotundae, Herb Gynostemmae Pentaphylli, carve leaf Pimpinella anisum Linn. etc.; Can excited cerebral neural Chinese medicine: Herba Ephedrae, the Radix Angelicae Dahuricae, Herba Xanthii grass, Radix Gentianae, Radix cocculi laurifolii, Semen Strychni, Cortex Periplocae etc.; There is the Chinese medicine of soporific function: Magnetitum, Os Draconis, Dens Draconis, succinum, Cinnabaris, Semen Ziziphi Spinosae, Radix Polygalae, Flos Albiziae etc.; Can the Chinese medicine of memory reinforcing: Rhizoma Acori Graminei, Fructus Alpiniae Oxyphyllae, Folium Ginkgo, Herba Herminii, Rhizoma Gastrodiae, Herba Boschniakiae Rossicae, Fructus Evodiae, the Radix Paeoniae Alba, the Rhizoma Anemarrhenae, Radix Panacis Quinquefolii etc.And many moneys Chinese medicine composition has also been proved the effect having memory and improve, as Yougui Wan, the kidney-Yang-Reinforcing Bolus, Bolus as a Kidney-Yin-Tonic etc. that six drugs containing rehmanniae Pelleting sanction, huperzine A capsule, Folium Ginkgo and Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd produce.
At present, on market, the product category of Improving memory power is a lot, but be subject to the restriction that long-term taking can produce some toxic and side effects, be suitable for the reasons such as crowd's scope is little, exploitation can long-term taking and the fundamentally health product of Improving memory power or medicine, become inexorable trend.
Summary of the invention
The application provides a kind of better effects if, toxic and side effects less, applicable crowd is Improving memory power Chinese medicine composition more widely, concrete, the application relates to a kind of Chinese medicine composition for Improving memory, and it comprises Radix Rehmanniae Preparata 15-35g, Rhizoma Dioscoreae 6-30g, Poria 6-20g, Semen Cuscutae 6-20g, Folium Ginkgo 6-16g.
The Chinese medicine composition of preferred described Improving memory comprises Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Poria 9g, Semen Cuscutae 12g, Folium Ginkgo 10g.
The Chinese medicine composition of preferred described Improving memory comprises Radix Rehmanniae Preparata 15-35g, Rhizoma Dioscoreae 6-30g, Fructus Corni 6-20g, Poria 6-20g, Cortex Moutan 6-20g, Rhizoma Alismatis 6-20g, Cortex Cinnamomi 1-10g, Fructus Lycii 6-20g, Semen Cuscutae 6-20g, Herba Epimedii 6-20g, Folium Ginkgo 6-16g.The Chinese medicine composition of further preferred described Improving memory comprises Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Fructus Corni 12g, Poria 9g, Cortex Moutan 9g, Rhizoma Alismatis 9g, Cortex Cinnamomi 6g, Fructus Lycii 12g, Semen Cuscutae 12g, Herba Epimedii 12g, Folium Ginkgo 10g.
The application also relates to a kind of preparation method of Chinese medicine composition of Improving memory, and it comprises the steps:
1) optional volatile oil is drawn: when comprising Fructus Corni, Cortex Moutan, Cortex Cinnamomi etc. in Chinese medicine composition, steam distillation is adopted to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) all the other medical materials of corresponding weight are taken, the water being equivalent to decoction pieces 10 times amount is added in extraction vessel, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filter, merge concentrated, then add the volatile oil of extraction, be prepared into basic, normal, high three concentration of Chinese medicine composition extract, amount to raw medicinal herbs and be respectively 0.53g/ml, 1.06g/ml, 2.12g/ml.
The application also relates to described Chinese medicine composition for the preparation of the purposes in the medicine of Improving memory, health product or food.
The medicine of described Improving memory, health product can add pharmaceutically acceptable adjuvant and be prepared into oral liquid, granule, electuary, tablet, capsule, pill etc.
The medicine of preferred described Improving memory, health product can add pharmaceutically acceptable adjuvant and be prepared into granule.
Beneficial effect:
The present invention is by optimizing known each Chinese medicine proportioning with Improving memory power, be that the Chinese medicine composition of the specific proportioning of monarch drug can improve scopolamine significantly and causes acquired dysmnesia model memory with Radix Rehmanniae Preparata, the effect of Improving memory is better than existing medicine, and not there is acute toxicity, genetoxic and chronic toxicity, the crowd of being suitable for is extensive, is suitable for Improving memory, amnesia and amnemonic control.
Accompanying drawing explanation
Fig. 1: water maze schematic diagram
Detailed description of the invention
Technical scheme of the present invention is further described by following embodiment.It is pointed out that following explanation is only illustrating the technical scheme that application claims is protected, any restriction not to these technical schemes.The content that protection scope of the present invention is recorded with appended claims is as the criterion.
Effect example 1: Chinese medicine composition is on the impact of mouse ability of learning and memory
1 experiment purpose: investigate Chinese medicine composition to the impact of mouse ability of learning and memory
2 experiment materials
2.1 instrument and equipment
DT-200 type mice diving tower instrument (Chengdu Tai Meng Science and Technology Ltd.)
BA-200 type mice keeps away dark instrument (Chengdu Tai Meng Science and Technology Ltd.)
Morris water maze (Chengdu Tai Meng Science and Technology Ltd.)
Rat shuttle box (Shanghai Ai Yan bio tech ltd HB-DSX2 type)
2.2 medicines and reagent
(1) Yougui Wan, the kidney-Yang-Reinforcing Bolus (Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd, the accurate word Z11021040 of traditional Chinese medicines), is configured to 1.76g/ml solution for standby;
(2) Bolus as a Kidney-Yin-Tonic (Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd, the accurate word Z11020735 of traditional Chinese medicines), is configured to 1.17g/ml solution for standby;
(3) Ginaton (Folium Ginkgo extract sheet) (Dr Willmar Schwabe, registration certificate number: H20090296), is configured to 7.8mg/ml solution for standby;
(4) huperzine A capsule (Chongqing Yao You pharmaceutical Co. Ltd, the accurate word H20030373 of traditional Chinese medicines), is configured to 0.02mg/ml solution for standby;
(5) scopolamine hydrobromide injection (Xuzhou Lai En pharmaceutcal corporation, Ltd, the accurate word H32022136 of traditional Chinese medicines);
(6) the application's Chinese medicine composition:
Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Fructus Corni 12g, Poria 9g, Cortex Moutan 9g, Rhizoma Alismatis 9g, Cortex Cinnamomi 6g, Fructus Lycii 12g, Semen Cuscutae 12g, Herba Epimedii 12g, Folium Ginkgo 10g, its preparation method is as follows:
1) volatile oil is drawn: adopt steam distillation to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) all the other medical materials of corresponding weight are taken, the water being equivalent to decoction pieces 10 times amount is added in extraction vessel, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filter, merge concentrated, concentrated, add the volatile oil of extraction again, be prepared into basic, normal, high three concentration of Chinese medicine composition extract, amount to raw medicinal herbs and be respectively 0.57g/ml, 1.14g/ml, 2.28g/ml.
2.3 animal
Male SPF level Kunming mouse, body weight 18-22 gram.Wistar rat, male, body weight 180-220g.
3 experimental programs and result
3.1 Jumping test
3.1.1 animal grouping and administration
Male SPF level Kunming mouse, 70, body weight 18-22 gram.Healthy kunming mice is divided into 7 groups at random, namely normal group, model group, huperzine A group (0.4mg/kgBW), Yougui Wan, the kidney-Yang-Reinforcing Bolus group (35.2g/kgBW), Bolus as a Kidney-Yin-Tonic group (23.4g/kgBW), Ginaton group (156mg/kgBW), the basic, normal, high dosage group of Chinese medicine composition (are respectively 15.33g/kgBW, 30.68g/kgBW, 61.34g/kgBW), often 10 are organized.Each group all by gastric infusion, administration volume is 20ml/kg, every day 1 time, successive administration 30 days.
3.1.2 experimental technique
Last administration is after 1 hour, and model group and each administration group mouse peritoneal injection scopolamine hydrobromide injection 5mg/kgBW set up mouse memory acquired disturbance model, and the normal saline of naive mice lumbar injection respective volume, carries out diving tower training after 10min.
During experiment, conform 3min animal to be put into reaction chamber (on platform, under platform), then by the copper grid in animal placing response case, passes to the alternating current of 36v immediately.Animal is shocked by electricity, and its normal reaction is rebound platform (insulator), to hide noxious stimulation.Most animals may skip on copper grid again or repeatedly, is subject to electric shock and snaps back again on platform.Be placed on by animal on the platform in reaction chamber, record mice jumps onto safety board required time (response time) after being shocked by electricity first, and in 5min, each Mus jumps off the errors number of platform, in this, as school grade.Carry out resurveying after 24h testing, mice is placed on platform, record the number of animals sum that each Mus first time jumps off number of shocks in the incubation period of platform, each Mus 3min and shocked by electricity, as recall tests achievement.
3.1.3 date processing and result judge
Incubation period, result was measurement data, available variance analysis, but first need carry out homogeneity test of variance by the program of variance analysis, and variance is neat, calculated F value, F value < F
0.05, conclusion: no significant difference between each group mean: F value>=F
0.05, P≤0.05, adds up with the comparative approach between two of mean between multiple experimental group and a matched group; Suitable variable transitions is carried out to the data of abnormal or heterogeneity of variance, after meeting normal state or variance and requiring together, adds up by the data after conversion; If do not reach normal state or the neat object of variance after variable transitions yet, use rank test instead and add up.
The number of animals of jumping off platform in errors number and 3min is numeration data, available X
2inspection, the total number of cases of four fold table is less than 40, or total number of cases is equal to or greater than 40, but when occurring that theoretical value is equal to or less than 1, should use exact method method instead.
If given the test agent group compares with matched group, incubation period obviously extends, and errors number or the number of animals of jumping off platform are obviously less than matched group, and difference has significance, and in above three indexs, any one index is positive, all can judge that this experiment is positive.
3.1.4 experimental result
Table 1 Chinese medicine composition causes memory acquisition disturbance model mice Jumping test to scopolamine
Note: in table, " * " represents to compare with model group to have aobvious person's difference (P < 0.05); " * * " represents to compare with model group to have and extremely shows person's difference (P < 0.01).
Compare with blank group, model group learning and memory latent time shortens, and errors number increases; Compared with model group, each positive control drug group, Chinese medicine composition group learning and memory latent time extend, and errors number reduces, and difference has statistical significance.Chinese medicine composition drug effect has dose-dependant, and high dose group effect is better than huperzine A group, and middle and high dosage group effect is better than Yougui Wan, the kidney-Yang-Reinforcing Bolus group, Bolus as a Kidney-Yin-Tonic group and Ginaton group.
3.2 darkness avoidance test
3.2.1 animal grouping and administration
Same 3.1.1
3.2.2 experimental technique
Last administration is after 1 hour, and model group and each administration group mouse peritoneal injection scopolamine hydrobromide injection 5mg/kgBW set up mouse memory acquired disturbance model, and the normal saline of naive mice lumbar injection respective volume, trains after 10min.
During experiment, mice face is put into bright room in hole dorsad, starts timer simultaneously.Animal enters darkroom through hole and is shocked by electricity, and timer stops automatically, takes out mice, records every Mus and meet with to entering darkroom the required time of shocking by electricity from putting into bright room, and this i.e. incubation period, training 5min, and records number of shocks in 5min.Recast test after 24h, records every animal and enters number of shocks in incubation period in darkroom and 5min.
3.2.3 date processing and result judge
Incubation period, result was measurement data, available variance analysis, but first need carry out homogeneity test of variance by the program of variance analysis, and variance is neat, calculated F value, F value < F
0.05, conclusion: no significant difference between each group mean: F value>=F
0.05, P≤0.05, adds up with the comparative approach between two of mean between multiple experimental group and a matched group; Suitable variable transitions is carried out to the data of abnormal or heterogeneity of variance, after meeting normal state or variance and requiring together, adds up by the data after conversion; If do not reach normal state or the neat object of variance after variable transitions yet, use rank test instead and add up.
Enter the number of animals entering darkroom in the number of times in darkroom and 5min in 5min and be enumeration data, available X
2inspection, the total number of cases of four fold table is less than 40, or total number of cases is equal to or greater than 40, but when occurring that theoretical value is equal to or less than 1, should use exact method method instead.
If the incubation period that given the test agent group mice enters darkroom is obviously longer than matched group, enter the number of animals entering darkroom in the errors number in darkroom or 5min in 5min and be less than matched group, and difference has significance, index any one of above three indexs is positive, all can judge that this experiment is positive.
3.2.4 experimental result
Table 2 Chinese medicine composition causes memory acquisition disturbance model mice step-through test to scopolamine
Note: in table, " * " represents to compare with model group to have significant difference (P < 0.05); " * * " represents to compare with model group to have pole significant difference (P < 0.01).
Compare with blank group, model group learning and memory latent time shortens, and errors number increases; Compared with model group, each positive control drug group, Chinese medicine composition group learning and memory latent time extend, and errors number reduces, and difference has statistical significance.Chinese medicine composition drug effect has dose-dependant, and high dose group effect is better than huperzine A group, and middle and high dosage group effect is better than Yougui Wan, the kidney-Yang-Reinforcing Bolus group, Bolus as a Kidney-Yin-Tonic group and Ginaton group.
3.3 water maze laboratory
3.3.1 animal grouping and administration
Same 3.1.1
3.3.2 experimental technique
Last administration is after 1 hour, and model group and each administration group mouse peritoneal injection scopolamine hydrobromide injection 5mg/kgBW set up mouse memory acquired disturbance model, and the normal saline of naive mice lumbar injection respective volume, trains after 10min.
Water maze test device (see Fig. 1) is a long 1.1m of being, the wide 0.7m of being, height is the brown lucite groove of 0.4m, there is ladder (platform, p) one end of groove, only has a road to arrive ladder, mice can be climbed up along it, and obtains rest.Be labyrinth-like in groove, mice can not see ladder, and there is cecum many places, is equipped with auto-induction apparatus; Automatic control monitor record data; Depth of water 10cm, water temperature (22 ± 1) DEG C, writing time 3min.
First inductivity training mice swimming labyrinth before test, can reach about 50% mice freely can swim out of water maze.During training, first mice is placed near P place and makes it climb up 2 times voluntarily, divided for 3 stages trained afterwards.In 1st stage: stop at A place with baffle plate, mice is trained from A place; In 2nd stage: stop at B place with baffle plate, mice is made to start training thus; 3rd stage: train by starting point S place.Each stage-training 2 times, carries out next stage training after rest 5min again.Mice rest 5min is made after whole training, being put near cat ladder makes it climb up 2 times voluntarily again, then directly S place is put in, record mice covering the race time (swimmingtime) and enter number of times and the errors number (numberoferror) of cecum, in 2min can not the person of swimming out of by 2min.The test result on modeling same day is school grade.24h after modeling, directly allows mice swimming labyrinth test, records its covering the race time, errors number equally, as recall tests achievement.
3.3.3 date processing and result judge
The time of reaching home belongs to measurement data, available variance analysis, but first need carry out homogeneity test of variance by the program of variance analysis, and variance is neat, calculates F value, F value < F
0.05, conclusion: no significant difference between each group mean: F value>=F
0.05, P≤0.05, adds up with the comparative approach between two of mean between multiple experimental group and a matched group; Suitable variable transitions is carried out to the data of abnormal or heterogeneity of variance, after meeting normal state or variance and requiring together, adds up by the data after conversion; If do not reach normal state or the neat object of variance after variable transitions yet, use rank test instead and add up.
Errors number and number of animals (percentage rate) two indexes of reaching home are enumeration data, available X
2inspection, the total number of cases of four forms is less than 40, or when total number of cases is equal to or greater than 40 but occurs that theoretical value is equal to or less than 1, should use exact method method instead.
Test group and matched group ratio, reach home time used or the errors number before reaching home of test group is obviously less than matched group, or the number of animals of reaching home in 2min is obviously more than matched group, and has significance through statistical test difference.Index any one of it is positive, can be judged to this experiment positive.
3.3.4 experimental result
Table 3 Chinese medicine composition causes memory acquisition disturbance model mice water maze laboratory to scopolamine
Note: in table, " * " represents to compare with model group to have significant difference (P < 0.05); " * * " represents to compare with model group to have pole significant difference (P < 0.01).
Compare with blank group, the mice covering the race time obviously extends, and errors number also obviously increases; Compared with model group, each positive controls and Chinese medicine composition group, the mice covering the race time obviously shortens, and errors number significantly reduces, and statistically has significant difference.Chinese medicine composition drug effect has dose-dependant, and high dose group effect is better than huperzine A group, and middle and high dosage group effect is better than Yougui Wan, the kidney-Yang-Reinforcing Bolus group, Bolus as a Kidney-Yin-Tonic group and Ginaton group.
3.4 shuttle back and forth experiment
3.4.1 animal grouping and administration
Male Wistar rat, 70, body weight 180-220 gram.Be divided into 7 groups at random, namely normal group, model group, huperzine A group (0.27mg/kgBW), Yougui Wan, the kidney-Yang-Reinforcing Bolus group (24.2g/kgBW), Bolus as a Kidney-Yin-Tonic group (16.1g/kgBW), Ginaton group (108mg/kgBW), the basic, normal, high dosage group of Chinese medicine composition (are respectively 10.58g/kgBW, 21.17g/kgBW, 42.33g/kgBW), often 10 are organized.Each group all by gastric infusion, administration volume is 10ml/kg, every day 1 time, successive administration 30 days.
3.4.2 experimental technique
Last administration is after 1 hour, and model group and each administration group mouse peritoneal injection scopolamine hydrobromide injection 5mg/kgBW set up mouse memory acquired disturbance model, and the normal saline of naive mice lumbar injection respective volume, trains after 10min.
Rat is put into any side of case, start after 20s to present light or buzz, continue 20s, in rear 10s, give electricity irritation (100V, 0.2mA, 50Hz, AC) simultaneously.Rat escapes to offside top after being shocked by electricity, and interrupts electric shock after blocking photocell.This is passive avoidance response, conditional stimulus is given before each electric shock, repeatedly after strengthening, namely rat acceptable conditions is jumped to offside after stimulating and is blocked photocell and escape electric shock, this is active avoidance response, every other day trains one time, often returns 50 times, after continuous training 4-5 returns, the active avoidance response rate of animal can reach more than 80%-90%.Index is analyzed as follows: animal number of shocks, the active avoidance time according to print result.Deconditioning, after 7 days, measures it and remembers the situation that disappears.
3.4.3 date processing and result judge
Animal active avoidance time and passive avoidance time are measurement data, available variance analysis.Experimental group active and/or passive avoidance time are significantly shorter than matched group, and difference has significance, be then judged to the positive.
3.4.4 experimental result
Table 4 Chinese medicine composition causes memory acquisition disturbance rat model to scopolamine and to shuttle back and forth experiment
Note: in table, " * " represents to compare with model group to have significant difference (P < 0.05); " * * " represents to compare with model group to have pole significant difference (P < 0.01).
Compare with blank group, rat active avoidance time shorten, number of shocks also obviously increases; Compared with model group, each positive controls and Chinese medicine composition group, rat active avoidance time shorten obviously extends, and number of shocks also significantly reduces, and statistically has significant difference.Chinese medicine composition drug effect has dose-dependant, and high dose group effect is better than huperzine A group, and middle and high dosage group effect is better than Yougui Wan, the kidney-Yang-Reinforcing Bolus group, Bolus as a Kidney-Yin-Tonic group and Ginaton group.
4 conclusions
Chinese medicine of the present invention obviously can improve scopolamine and cause memory acquisition disturbance model memory, and Chinese medicine composition effect of the present invention is suitable with huperzine A effect, and middle and high dosage group is obviously better than Yougui Wan, the kidney-Yang-Reinforcing Bolus, Bolus as a Kidney-Yin-Tonic group and Ginaton group.
Effect example 2: the acute toxicity test of Chinese medicine composition
1 test objective: whether observe Chinese medicine composition has Acute Toxicity to body
2 experiment materials
2.1 medicines and reagent
Chinese medicine composition extract
2.2 instrument and equipment
Metal gastric perfusion needle, syringe
2.3 animal
Kunming mouse, SPF level, body weight 18-22g, male and female half and half
3 experimental techniques
Animal is observed 5 days after buying, fasting before administration (not water restriction) 16h, with the Cmax of Chinese medicine composition extract (4.77g/ml) and maximum administration volume (4.0ml/100gBw) gastric infusion, blank group gives the normal saline of respective volume, Continuous Observation 14 days, the mice that mice and the observation of death expire is dissected, observes each change organizing internal organs.4 interpretations of result and judgement
Continuous Observation 14 days, if animal does not occur death, then thinks that Chinese medicine composition extract is greater than study dosage to Kunming mouse per os acute toxicity dosage.
5 experimental results
Continuous Observation 14 days, none animal of result occurs dead, administration group mice searches for food, drink water, the mental status with activity situation compared with blank group, without significant change.
To observation mice after 14 days, cut open inspection and find, respectively organize internal organs, after one's own heart, liver, spleen, harmonization of the stomach kidney etc. and naive mice internal organs thing notable difference.
6 experiment conclusion:
The maximum tolerated dose of Chinese medicine composition extract Kunming mouse gastric infusion is greater than 190g/kgBw (being converted into crude drug amount), is equivalent to 106 times of people's consumption, can judges that Chinese medicine composition belongs to nontoxic level.
Effect example 3: the genetic toxicity test of Chinese medicine composition
1 test objective: whether observe Chinese medicine composition has Genotoxic Effect to body
2 experiment materials
2.1 medicines and reagent
Chinese medicine composition extract
1%-2% eosin stains liquid
2.2 instrument and equipment
Metal gastric perfusion needle, syringe
Biological microscope
2.3 animal
Male mice in kunming, SPF level, body weight 18-22g
3 experimental techniques
50 male mice in kunming, be divided into 5 groups at random, Chinese medicine composition group establishes 3 dosage groups, take maximum tolerated dose as high dose group, 1/2 of maximum tolerated dose is middle dosage group, 1/4 of maximum tolerated dose is low dose group, separately establishes 1 positive controls and feminine gender (solvent) matched group, often organizes 10.Positive controls selects cyclophosphamide, and gastric infusion dosage is 40mg/kgBW.Mouse stomach volume is 0.2ml/10gBW, every day 1 time, continuous 5 days, every minor tick 24h.Within the 35th day after gastric infusion first, put to death mice with cervical dislocation, win both sides epididymis, be placed with in the small beaker of appropriate normal saline (about 1mL) or put into the plate filling 2mL normal saline.With eye scissors, epididymis is longitudinally cut 1-2 cutter, static 3-5min, shakes gently.Filter with four layers of lens paper or synthetic fibers blood mesh bag, suction strainer liquid smear.After air drying, fix more than 5min with methanol dry, with 1%-2% eosin stains liquid 1h, gently rush with water, dry.
4 interpretations of result and judgement
Sperm deformity, is mainly manifested in head, is secondly afterbody.Lopsided type can be divided into Wugou, banana-shaped, fat head, amorphous, tail folding, double end, two tails etc.
Each dosage group should compare respectively at corresponding negative control group, as evaluated sperm deformity positive criteria by Wilcoson rank test method is, abnormal rate be at least negative control group times amount or through statistics significance, and have dose-response relationship.
General negative control group cacospermia rate is 0.8%-3.4%.
5 experimental results
Table 1 Chinese medicine composition extract mouse inbred strain result
Note: in table, " * " represents to compare with negative control group to have significant difference (P < 0.05); " * * " represents to compare with negative control group to have significant difference (P < 0.01).
6 experiment conclusion:
Chinese medicine composition extract mouse sperm deformity rate in 95.24-381g/kgBW dosage range compares with negative control group without the significance difference opposite sex, can judge Chinese medicine composition thing genetoxic.
Effect example 4: the chronic toxicity test of Chinese medicine composition
1 test objective: whether observe Chinese medicine composition has chronic toxic effect to body
2 experiment materials
2.1 medicines and reagent
Chinese medicine composition extract
2.2 instrument and equipment
Metal gastric perfusion needle, syringe
2.3 animal
SD rat, 6-8 week age, SPF level, male and female half and half
3 experimental techniques
3.1 animal grouping and administrations
88 rats, experimental animal healthy after quarantine is observed is divided into 4 groups at random, often organizes 22, male and female half and half.Chinese medicine composition group establishes 3 dosage groups, and with 1/4 of maximum tolerated dose for high dose group, 1/8 of maximum tolerated dose is middle dosage group, and 1/16 of maximum tolerated dose is low dose group, separately establishes feminine gender (solvent) matched group, often organizes 10.
Route of administration is per os, every day gastric infusion 1 time, continuous 90 days.
3.2 clinical observations and inspection
3.2.1 clinical observation
Duration of test observes outward appearance sign, the behavioral activity and feces shape etc. of 1 all test group and control animals every day, there are time of ANOMALOUS VARIATIONS and situation etc. in record animal, as found, animal has death or the gross anatomy of doing in time of being at death's door to observe, and leaves and takes that organ-tissue sample is standby does histopathologic examination.Observe every day and record the food-intake of animal, weighing the weight of animals 1 time per weekend.
3.2.2 clinical examination
1) hematological indices inspection: during off-test, often group takes 12 rats (male and female half and half) blood sample, detects with SysmexpocH-lOOi type entry automatic hemacytometer.Index for examination have red blood cell count(RBC) (RBC), leukocyte (WBC) count and classification, hemoglobin (HGB), platelet count (PET), hematid specific volume (HCT) etc.
2) blood biochemical (liver function) checks: often organize during off-test and gather respectively and be separated 12 rats (male and female half and half) serum, detect with Rx-daytona automatic clinical chemistry analyzer.Detect the indexs such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkali phosphatase (ALP), blood urea nitrogen (BUN), total protein (TP), albumin (ALB), blood glucose (GLU), total bilirubin (TBIL), T-CHOL (cHo), acetylcholine esterase (ChE).
3) urine (renal function) checks: before off-test 2d often group get 12 rats (male and female half and half) and put respectively in people's metabolic cage and raise, collect 24h urine, detect with ClinitekStatus full automatic urine analyzer.Testing index has outward appearance, acid-base value (pH), occult blood (BLD), albumen (PRo), glucose in urine (U.GLU) and precipitate microscopy etc.
4) organ weights measures: the animal of putting to death all test group and matched group during off-test, weighs the body weight of every animal and the heart, liver, spleen, lung, kidney, brain, stomach, uterus respectively, the Main Tissues internal organs such as testis.
5) pathological examination: to all animals of putting to death when duration of test death and off-test, all carries out anatomic observation and leaves and takes organ-tissue sample for making pathological examination.
4 interpretations of result and judgement
Data transformations and statistical analysis is carried out with SPSS (11.0) software.During analysis, first homogeneity test of variance is carried out to data, as neat in sent out difference, adopt one factor analysis of variance totally to compare, note abnormalities and carry out comparing between two between multiple dose group with a matched group mean by Dunnett method again.If heterogeneity of variance, then suitable quantity conversion is carried out to initial data, after meeting homogeneity test of variance, add up by the data after conversion; If do not reach the neat object of variance after variable transitions yet, use rank test instead and add up, find overall more variant, then adopt and do not require that the TamhanesT2 of homogeneity of variance inspection is compared between two.
5 experimental results
5.1 Chinese medicine compositions are on the impact of hematological indices
To the observed result of the hematological indices such as RBC, WBC, L, M, N, HGB, PLT, HCT in table 1, the indices of each dosage group of male and female rat compares with matched group all without significant difference (P > 0.05).When showing that Chinese medicine composition dosage reaches 62.5g/kgBW, obvious impact is not all produced on surveyed every hematological indices.
Table 1 Chinese medicine composition is on the impact of rat blood index
Note: in table, " * " represents to compare with negative control group to have significant difference (P < 0.05); " * * " represents to compare with negative control group to have significant difference (P < 0.01).
5.2 impacts on liver function
With the project such as serum AST, ALT, ALP, TP, ALB, GLU, T-BIL, CHO, ChE for observation index, testing result is in table 2, male rat T-BIL with CHO, female rats AST are variant compared with blank group with ChE, but difference does not have statistical significance, do not have a dose-dependant characteristic yet.Experimental result shows, Chinese medicine composition is fed for a long time and can not be produced material impact to rat liver.
Table 2 Chinese medicine composition is to rat liver function effect
Note: in table, " * " represents to compare with negative control group to have significant difference (P < 0.05); " * * " represents to compare with negative control group to have significant difference (P < 0.01).
5.3 Chinese medicine compositions are to renal function
Rat urine BUN, CRE are observed.Experimental result is in table 3, and compared with matched group, rat BUN and CRE has certain change, but change is not remarkable, does not have statistical significance, and without dose-response relationship.Experimental result shows, Chinese medicine composition is fed for a long time and can not be produced material impact to Renal Function.
Table 3 Chinese medicine composition affects Renal Function in Rats
Note: in table, " * " represents to compare with negative control group to have significant difference (P < 0.05); " * * " represents to compare with negative control group to have significant difference (P < 0.01).
5.4 Chinese medicine compositions affect organ coefficient
To the organ coefficient measurement result of the rat heart, liver, spleen, lung, kidney, brain major organs in table 4, compared with blank group, the organ coefficient of internal organs is respectively organized not have significant difference.
Table 4 Chinese medicine composition is on the impact of organ coefficient
Note: in table, " * " represents to compare with negative control group to have significant difference (P < 0.05); " * * " represents to compare with negative control group to have significant difference (P < 0.01).
6 experiment conclusion:
Chinese medicine composition is fed rat blood biochemical indicator, hepatic and renal function for a long time, is respectively organized internal organs to produce material impact.
Embodiment 1: a kind of Chinese medicine oral liquid of Improving memory
Radix Rehmanniae Preparata 35g, Rhizoma Dioscoreae 30g, Poria 20g, Semen Cuscutae 20g, Folium Ginkgo 16g
Take the medical material of corresponding weight, in extraction vessel, add the water being equivalent to medical material 10 times amount, after soaking 30min, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filter, merge concentrated, be prepared into Chinese medicine composition extract, amounting to raw medicinal herbs is 2.12g/ml.
Medicinal liquid lets cool, stock solution embedding, irradiation sterilization, and after inspection, packed products, obtains the Chinese medicine oral liquid of Improving memory power.
Embodiment 2: a kind of Chinese medicine granules of Improving memory
Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Fructus Corni 12g, Poria 9g, Cortex Moutan 9g, Rhizoma Alismatis 9g, Cortex Cinnamomi 6g, Fructus Lycii 12g, Semen Cuscutae 12g, Herba Epimedii 12g, Folium Ginkgo 10g.
1) volatile oil is drawn: adopt steam distillation to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) all the other medical materials of corresponding weight are taken, the water being equivalent to decoction pieces 10 times amount is added in extraction vessel, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filter, merge concentrated, when obtaining 50-60 DEG C, relative density is the clear paste powder of 1.20-1.32, add the acceptable filler of pharmacy, as starch, lactose, the mixings such as microcrystalline Cellulose, add starch slurry or polyvidone or sodium carboxymethyl cellulose wet granulation, granulate after dry, volatile oil is evenly sprayed on granule, put hermetic container and absorb 48 hours, packaging, obtain the Chinese medicine granules of Improving memory.
Embodiment 3: a kind of Chinese medicine capsules of Improving memory
Granule embodiment 2 prepared loads gelatine capsule, obtains the Chinese medicine capsules of Improving memory.
Embodiment 4: a kind of Chinese medicinal tablet of Improving memory
Tabletting add magnesium stearate in granule prepared by embodiment 2 after, obtains the Chinese medicinal tablet of Improving memory.
Embodiment 5: a kind of instant particles as Chinese medicine of Improving memory
Radix Rehmanniae Preparata 15g, Rhizoma Dioscoreae 30g, Fructus Corni 20g, Poria 20g, Cortex Moutan 20g, Rhizoma Alismatis 20g, Cortex Cinnamomi 10g, Fructus Lycii 20g, Semen Cuscutae 20g, Herba Epimedii 20g, Folium Ginkgo 16g, its preparation method is as follows:
1) volatile oil is drawn: adopt steam distillation to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) all the other medical materials of corresponding weight are taken, the water being equivalent to decoction pieces 10 times amount is added in extraction vessel, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filter, merge concentrated, when obtaining 50-60 DEG C, relative density is the clear paste powder of 1.20-1.32, add the acceptable filler of pharmacy, as starch, lactose, the mixings such as microcrystalline Cellulose, add starch slurry or polyvidone or sodium carboxymethyl cellulose wet granulation, granulate after dry, again granule is rubbed with the hands broken, volatile oil is evenly sprayed on rub with the hands broken after granule on, put hermetic container and absorb 48 hours, packaging, obtain the instant particles as Chinese medicine of Improving memory.
Embodiment 6: a kind of medicine pill of Improving memory
Radix Rehmanniae Preparata 35g, Rhizoma Dioscoreae 30g, Fructus Corni 20g, Poria 20g, Cortex Moutan 15g, Rhizoma Alismatis 15g, Cortex Cinnamomi 10g, Fructus Lycii 15g, Semen Cuscutae 15g, Herba Epimedii 15g, Folium Ginkgo 10g, its preparation method is as follows:
1) volatile oil is drawn: adopt steam distillation to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) take all the other medical materials of corresponding weight, in extraction vessel, add the water being equivalent to decoction pieces 10 times amount, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filters, merge concentrated, concentrated, be the clear paste powder of 1.20-1.32 relative to density when obtaining 50-60 DEG C, add volatile oil after pulverizing, mix to obtain medicated powder.Mel boils to temperature 119 DEG C, after by temperature to 60-70 DEG C, add medicated powder be in harmonious proportion knead, obtain the medicine pill of Improving memory.
The present invention is that the Chinese medicine composition of the specific proportioning of monarch drug can improve scopolamine significantly and causes the acquired dysmnesia of mice with Radix Rehmanniae Preparata, and pharmacological action presents dose-dependent characteristic substantially.Further, Chinese medicine composition high dose group is better than huperzine A, and middle and high dosage group is better than Yougui Wan, the kidney-Yang-Reinforcing Bolus, Bolus as a Kidney-Yin-Tonic and Ginaton.Chinese medicine composition of the present invention does not have acute toxicity, genetoxic and chronic toxicity, is suitable for memory improvement amnesia and amnemonic control.
Content of the present invention merely illustrates some claimed specific embodiments; one of them or more described technical characteristic can be combined with arbitrary one or more technical scheme in technical scheme; these technical schemes obtained through combination also in the application's protection domain, just as these technical schemes obtained through combination in the disclosure of invention concrete record.
Claims (9)
1. for a Chinese medicine composition for Improving memory, it is characterized in that, described compositions is prepared from by following crude drug: Radix Rehmanniae Preparata 15-35g, Rhizoma Dioscoreae 6-30g, Poria 6-20g, Semen Cuscutae 6-20g, Folium Ginkgo 6-16g.
2. Chinese medicine composition according to claim 1, is characterized in that Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Poria 9g, Semen Cuscutae 12g, Folium Ginkgo 10g.
3. Chinese medicine composition according to claim 1, is characterized in that, can also add in described compositions: Fructus Corni 6-20g, Cortex Moutan 6-20g, Rhizoma Alismatis 6-20g, Cortex Cinnamomi 1-10g, Fructus Lycii 6-20g, Herba Epimedii 6-20g.
4. Chinese medicine composition according to claim 3, is characterized in that, Fructus Corni 12g, Cortex Moutan 9g, Rhizoma Alismatis 9g, Cortex Cinnamomi 6g, Fructus Lycii 12g, Herba Epimedii 12g.
5. the Chinese medicine composition according to any one of claim 3-4, is characterized in that described Chinese medicine composition is prepared from by following crude drug: Radix Rehmanniae Preparata 24g, Rhizoma Dioscoreae 12g, Fructus Corni 12g, Poria 9g, Cortex Moutan 9g, Rhizoma Alismatis 9g, Cortex Cinnamomi 6g, Fructus Lycii 12g, Semen Cuscutae 12g, Herba Epimedii 12g, Folium Ginkgo 10g.
6. the Chinese medicine composition according to any one of claim 1-5, is characterized in that, described Chinese medicine composition adds pharmaceutically acceptable adjuvant agent and is prepared into oral liquid, granule, electuary, tablet, capsule, pill.
7. Chinese medicine composition according to claim 6, it is characterized in that, described Chinese medicine composition is granule.
8. a preparation method for the Chinese medicine composition described in any one of claim 1-6, it comprises the steps:
1) optional volatile oil is drawn: when there is Fructus Corni, Cortex Moutan, Cortex Cinnamomi etc. in Chinese medicine composition, steam distillation is adopted to extract volatile ingredient contained in Fructus Corni, Cortex Moutan, Cortex Cinnamomi, extraction process is: take the Fructus Corni of corresponding weight, Cortex Moutan, Cortex Cinnamomi, add the water soaking 1 hour of 10 times of medical material amounts, volatile oil is extracted 4 hours with volatile oil distillator, for subsequent use;
2) take all the other medical materials of corresponding weight, in extraction vessel, add the water being equivalent to decoction pieces 10 times amount, after immersion 30min together with the remaining medical material of volatile oil is drawn, decoct 30min, 2 times, filter, merging filtrate, or with 10 times amount ethanol or Diluted Alcohols, soak 30min, reflux, extract, 2 times, each 1 hour, filters, merging filtrate, concentrated, then add the volatile oil of extraction and pharmaceutically acceptable adjuvant is prepared into corresponding dosage form, obtain the Chinese medicine composition of Improving memory.
9. the Chinese medicine composition described in an any one of claim 1-6 is for the preparation of the purposes in the medicine of Improving memory, health product or food.
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| KR20110110574A (en) * | 2010-04-01 | 2011-10-07 | (주)아모레퍼시픽 | Composition for improving study skills |
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