CN107737207B - Pharmaceutical composition with antidepressant effect and preparation method and application thereof - Google Patents
Pharmaceutical composition with antidepressant effect and preparation method and application thereof Download PDFInfo
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- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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Abstract
The invention discloses an antidepressant pharmaceutical composition, a preparation method and application thereof. The pharmaceutical composition is prepared from 10-70 parts by weight of ginseng, 30-120 parts by weight of rhizoma acori graminei, 50-220 parts by weight of poria cocos, 15-50 parts by weight of polygala tenuifolia, 0.5-2 parts by weight of rosemary volatile oil and 0.5-2 parts by weight of rose volatile oil. The pharmaceutical composition has a remarkable antidepressant effect.
Description
Technical Field
The invention relates to a pharmaceutical composition with an antidepressant effect, and also relates to a preparation method and application of the composition.
Background
The leading good forgetting recipe was originally found in Tang Dynassia Sun Simiao (prescription of Qian jin of Bei Ji.) Vol (fourteenth Small intestine) prepared from cortex et radix Polygalae, Ginseng radix, Poria, and rhizoma Acori Calami. The recipe of good forgetting is mainly derived from many prescriptions, such as Ding Zhi Wan, Ding Zhi Xiao Wan, Bu Xin Tang, Dou xi san, Yuan Zhi san and Zhuang Yuan Wan. At present, the menstruation prescriptions are developed to be applied to the treatment of various diseases, such as anxiety, amnesia, depression, abnormal speech, senile dementia and the like. The traditional preparation method of the main good forgetting prescription and related derived prescriptions comprises the steps of crushing and preparing into powder and pills, or directly decocting with water to prepare into a water decoction.
In modern researches on the main good forgetting formula and related formulas, the traditional Chinese medicine preparation is mainly a new formula obtained by optimizing the proportion of the components of the medicine and adding or subtracting the medicinal flavor on the basis of the main good forgetting formula. The preparation method of the basic prescription of the medicine generally comprises a direct medicinal material crushing method, a water extraction and alcohol precipitation method and the like; in addition, some of the prior art also include a step of extracting volatile oil from the grass-leaved sweetflag. The prior art has a great emphasis on treating related diseases, but the overall effect is still to be improved.
Disclosure of Invention
The invention aims to provide a medicine composition which is obtained based on a main good forgetting prescription and has more remarkable antidepressant effect.
The invention also aims to provide a preparation method of the pharmaceutical composition, and the pharmaceutical composition obtained by the preparation method has better anti-depression effect.
Still another object of the present invention is to provide a pharmaceutical preparation containing the above pharmaceutical composition.
The invention also aims to provide the application of the pharmaceutical composition.
The purpose of the invention is realized by the following technical scheme:
a pharmaceutical composition for resisting depression is prepared from 10-70 parts by weight of ginseng, 30-120 parts by weight of rhizoma acori graminei, 50-220 parts by weight of poria cocos, 15-50 parts by weight of polygala tenuifolia, 0.5-2 parts by weight of rosemary volatile oil and 0.5-2 parts by weight of rose volatile oil.
According to the pharmaceutical composition, the pharmaceutical composition is preferably prepared from 20-60 parts by weight of ginseng, 40-100 parts by weight of rhizoma acori graminei, 60-200 parts by weight of poria cocos, 18-40 parts by weight of polygala tenuifolia, 1-2 parts by weight of rosemary volatile oil and 0.5-1 part by weight of rose volatile oil.
The pharmaceutical composition according to the present invention is preferably prepared from the following components:
20 parts of ginseng, 100 parts of rhizoma acori graminei, 200 parts of poria cocos, 20 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
60 parts of ginseng, 40 parts of rhizoma acori graminei, 60 parts of poria cocos, 40 parts of polygala tenuifolia, 1 part of rosemary volatile oil and 0.5 part of rose volatile oil; or
32 parts of ginseng, 40 parts of rhizoma acori graminei, 160 parts of poria cocos, 40 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
32 parts of ginseng, 80 parts of acorus gramineus, 160 parts of poria cocos, 32 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
20 parts of ginseng, 40 parts of acorus gramineus, 70 parts of poria cocos, 18 parts of polygala tenuifolia, 1 part of rosemary volatile oil and 0.5 part of rose volatile oil.
In the invention, the rosemary volatile oil is obtained by extracting rosemary by a steam distillation supercritical extraction method. The volatile oil of flos Rosae Rugosae is extracted from flos Rosae Rugosae by supercritical extraction with steam distillation.
The invention also provides a preparation method of the pharmaceutical composition. According to one embodiment of the present invention, the preparation method of the pharmaceutical composition comprises the steps of:
(1) extracting Ginseng radix, rhizoma Acori Graminei, Poria and cortex et radix Polygalae with water as solvent to obtain water extractive solution, concentrating the water extractive solution, and drying to obtain water extract;
(2) clathrating the rosemary volatile oil and the rose volatile oil with cyclodextrin to obtain a volatile oil clathrate;
(3) and uniformly mixing the water extract and the volatile oil clathrate compound to obtain the pharmaceutical composition.
According to another embodiment of the present invention, the preparation method of the pharmaceutical composition comprises the steps of:
(1') extracting volatile oil from ginseng, rhizoma acori graminei and polygala tenuifolia to obtain mixed volatile oil and dregs;
(2') clathrating the mixed volatile oil, the rosemary volatile oil and the rose volatile oil with cyclodextrin to obtain a volatile oil clathrate;
(3') extracting the residue and Poria cocos wolf with water as solvent to obtain water extract, concentrating the water extract, and drying to obtain water extract;
(4') uniformly mixing the volatile oil inclusion compound and the water extract to obtain the medicine composition.
In the step (1'), the ginseng, the acorus gramineus soland and the polygala tenuifolia are respectively extracted with volatile oil and then uniformly mixed to obtain the mixed volatile oil, or the ginseng, the acorus gramineus soland and the polygala tenuifolia are mixed to extract the volatile oil to obtain the mixed volatile oil. In the invention, the method for extracting the volatile oil can be a steam distillation method or a supercritical extraction method. According to one embodiment of the invention, the method for extracting volatile oil is a steam distillation method comprising: adding water into the ginseng, the acorus gramineus soland and the polygala tenuifolia for steam distillation, wherein the water adding amount is 8-15 times of the total weight of the ginseng, the acorus gramineus soland and the polygala tenuifolia, and is preferably 10-14 times of the total weight of the ginseng, the acorus gramineus soland and the polygala tenuifolia; distilling for 4-7 hours, preferably for 5-6 hours to obtain mixed volatile oil and an extracting solution; filtering the extractive solution to obtain medicinal liquid and residue. According to one embodiment of the present invention, the method for extracting volatile oil is a supercritical extraction method comprising: extracting the ginseng, the rhizoma acori graminei and the polygala tenuifolia by adopting a supercritical carbon dioxide extraction method to obtain volatile oil, wherein the extraction temperature is 45-65 ℃, and preferably 50-60 ℃; the extraction pressure is 25-32 mPa, preferably 28-30 mPa; the extraction time is 60-120 min, preferably 80-100 min, and the mixed volatile oil is obtained.
In the step (2'), the mixed volatile oil, the rosemary volatile oil and the rose volatile oil can be uniformly mixed after being respectively included by cyclodextrin, so as to obtain the volatile oil inclusion compound. According to a preferred embodiment of the invention, the mixed volatile oil, the rosemary volatile oil and the rose volatile oil are uniformly mixed and then are included by cyclodextrin, so as to obtain a volatile oil inclusion compound; preferably, the cyclodextrin is beta-cyclodextrin, and the dosage of the beta-cyclodextrin is 2-6 times of the total weight of the mixed volatile oil, the rosemary volatile oil and the rose volatile oil, and is preferably 3.5-5 times.
In the step (3'), the extraction method may be a decoction method, a reflux extraction method or an ultrasonic extraction method, and is preferably a decoction method or a reflux extraction method. In the invention, preferably, the decoction dregs and the poria are mixed and then extracted by adding water for 1-4 times, preferably 2-3 times; the water amount is 5-15 times of the total weight of the ginseng, the rhizoma acori graminei, the polygala tenuifolia and the poria cocos, and preferably 6-8 times of the total weight of the ginseng, the rhizoma acori graminei, the polygala tenuifolia and the poria cocos; the extraction time is 0.5-3 hours, preferably 1-2 hours, and the water extract is obtained. According to one embodiment of the invention, the decoction dregs and the poria cocos are mixed and then extracted with water for 2-3 times, the water addition amount is 6-8 times of the total weight of the ginseng, the rhizoma acori graminei, the polygala tenuifolia and the poria cocos, and the extraction time is 1-2 hours, so that the water extraction solution is obtained.
According to one embodiment of the present invention, in the step (1'), the method for extracting volatile oil is steam distillation; step (3') further comprises: and uniformly mixing the water extract and the liquid medicine to obtain a mixed solution, concentrating the mixed solution, and drying to obtain the water extract.
According to one embodiment of the present invention, the preparation method of the pharmaceutical composition comprises: extracting volatile oil from Ginseng radix, rhizoma Acori Graminei and cortex et radix Polygalae by steam distillation, adding water 12 times of the total weight of Ginseng radix, rhizoma Acori Graminei and cortex et radix Polygalae, and extracting for 5 hr to obtain mixed volatile oil and extractive solution; filtering the extractive solution to obtain medicinal liquid and residue; uniformly mixing the mixed volatile oil, the rosemary volatile oil and the rose volatile oil, adding beta-cyclodextrin for inclusion, drying at 60 ℃, and crushing to obtain a volatile oil inclusion compound, wherein the dosage of the beta-cyclodextrin is 4 times of the total weight of the mixed volatile oil, the rosemary volatile oil and the rose volatile oil; mixing the residues with Poria, decocting in water for 2 times (8 times of the total weight of Ginseng radix, rhizoma Acori Graminei, cortex et radix Polygalae and Poria for 1 hr), filtering, mixing the filtrate with the above medicinal liquid, concentrating, drying, and pulverizing to obtain water extract; and uniformly mixing the water extract and the volatile oil clathrate compound to obtain the pharmaceutical composition.
According to another embodiment of the present invention, the preparation method of the pharmaceutical composition comprises: extracting Ginseng radix, rhizoma Acori Graminei and cortex et radix Polygalae with supercritical carbon dioxide at 50 deg.C under 28mPa for 90min to obtain mixed volatile oil and residue; uniformly mixing the mixed volatile oil, the rosemary volatile oil and the rose volatile oil, adding beta-cyclodextrin for inclusion, drying at 60 ℃, and then crushing to obtain a volatile oil inclusion compound, wherein the dosage of the beta-cyclodextrin is 5 times of the total weight of the mixed volatile oil, the rosemary volatile oil and the rose volatile oil; mixing the residues with Poria, decocting in water for 2 times (8 times of the total weight of Ginseng radix, rhizoma Acori Graminei, cortex et radix Polygalae and Poria for 1 hr), filtering to obtain water extractive solution, concentrating the water extractive solution, drying and pulverizing to obtain water extract; and uniformly mixing the water extract and the volatile oil clathrate compound to obtain the pharmaceutical composition.
The invention also provides a medicinal preparation for resisting depression, which comprises the medicinal composition and pharmaceutically acceptable auxiliary materials. The dosage form of the pharmaceutical preparation is not limited, but is preferably oral dosage form, such as soft capsules, hard capsules, pills, tablets, granules, powder and the like. The pharmaceutically acceptable auxiliary materials include, but are not limited to, starch, dextrin, sucrose, microcrystalline cellulose, lactose, polyethylene glycol 4000, polyethylene glycol 6000, vegetable oil, beeswax and the like.
The invention also provides application of the pharmaceutical composition in preparing antidepressant drugs.
In the invention, the rosemary volatile oil and the rose volatile oil with proper proportion are added on the basis of the ginseng, the grassleaf sweelflag rhizome, the tuckahoe and the polygala root, thereby enhancing the anti-depression curative effect of the pharmaceutical composition; compared with the traditional preparation method, the invention extracts the volatile oil components in the medicinal materials, and particularly extracts the volatile oil components of ginseng and polygala tenuifolia in addition to the rhizoma acori graminei rich in volatile oil, so that the antidepressant effect of the pharmaceutical composition is obviously superior to that of the pharmaceutical composition prepared by the traditional method (such as a crude drug crushing method).
Detailed Description
The present invention will be further described with reference to the following specific examples, but the scope of the present invention is not limited thereto.
In the present invention, ginseng is a dried root and rhizome of Panax ginseng C.A.Mey. Polygala tenuifolia Willd is dried root of Polygala tenuifolia Willd or Polygala sibirica L. Rhizoma Acori Graminei is dried rhizome of Acorus tatarinowii Schott of Araceae. Poria is dried sclerotium of Wolf of Poria cocos (Schw.) belonging to Polyporaceae. Rosemary is derived from Rosmarinus officinalis L. flos Rosae Rugosae of Rosmarinus of Labiatae, and is dried flower bud of Rosa rugosa Thunb. of Rosaceae.
Example 1
Taking 200g of ginseng, 1000g of rhizoma acori graminei and 200g of polygala tenuifolia, uniformly mixing, crushing into coarse powder, extracting volatile oil by adopting a steam distillation method, adding 19.6kg of water, extracting for 6 hours to obtain mixed volatile oil and an extracting solution, and filtering the extracting solution to obtain liquid medicine and dregs of a decoction; uniformly mixing the mixed volatile oil with 20g of rosemary volatile oil and 10g of rose volatile oil, adding beta-cyclodextrin which is 5 times of the total weight of the volatile oil for inclusion, drying at 55-60 ℃, and crushing to obtain a volatile oil inclusion compound; mixing the residue with 2000g Poria, decocting in water for 2 times (30.6 kg for each time and 1 hr for each time), filtering, mixing the filtrate with the above medicinal liquid, concentrating, drying, and pulverizing to obtain water extract; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition.
Example 2
Mixing Ginseng radix 600g, rhizoma Acori Graminei 400g and cortex et radix Polygalae 400g, pulverizing into coarse powder, extracting volatile oil by steam distillation with water amount of 16.8kg for 5 hr to obtain mixed volatile oil and extractive solution, and filtering the extractive solution to obtain medicinal liquid and residue; uniformly mixing the mixed volatile oil with 10g of rosemary volatile oil and 5g of rose volatile oil, adding beta-cyclodextrin which is 4 times of the total weight of the volatile oil for inclusion, drying at 55-60 ℃, and crushing to obtain a volatile oil inclusion compound; mixing the residue with 600g Poria, adding water, reflux-extracting for 2 times (each time with 16kg water and 1 hr extraction time), filtering, mixing the filtrate with the above medicinal liquid, concentrating, drying, and pulverizing to obtain water extract; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition.
Example 3
Taking 320g of ginseng, 400g of rhizoma acori graminei and 400g of polygala tenuifolia, uniformly mixing, crushing into coarse powder, placing the coarse powder in an extraction kettle of a supercritical carbon dioxide extraction apparatus, extracting at the temperature of 55 ℃, under the extraction pressure of 30mPa for 90min, and obtaining mixed volatile oil and dregs; uniformly mixing the mixed volatile oil with 20g of rosemary volatile oil and 10g of rose volatile oil, adding beta-cyclodextrin which is 6 times of the total weight of the volatile oil for inclusion, drying at 55-60 ℃, and crushing to obtain a volatile oil inclusion compound; mixing the residue with 1600g Poria, decocting in water for 3 times (water amount per time is 27.2 kg) and each time for 1 hr to obtain water extractive solution, concentrating, drying, and pulverizing to obtain water extract; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition.
Example 4
Taking 320g of ginseng, 800g of rhizoma acori graminei and 320g of polygala tenuifolia, uniformly mixing, crushing into coarse powder, placing the coarse powder in an extraction kettle of a supercritical carbon dioxide extraction apparatus, extracting at the temperature of 50 ℃, under the extraction pressure of 28mPa for 90min, and obtaining mixed volatile oil and dregs; uniformly mixing the mixed volatile oil with 20g of rosemary volatile oil and 10g of rose volatile oil, adding beta-cyclodextrin which is 5 times of the total weight of the volatile oil for inclusion, drying at 55-60 ℃, and crushing to obtain a volatile oil inclusion compound; mixing the residue with 1600g Poria, decocting in water for 2 times (water amount per time is 24.32 kg) and each time for 1 hr to obtain water extractive solution, concentrating, drying, and pulverizing to obtain water extract; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition.
Example 5
Extracting Ginseng radix 200g, rhizoma Acori Graminei 400g, Poria 700g and cortex et radix Polygalae 180g with water as solvent for 2 times, each time adding water 14.8kg, and extracting for 2 hr and 1 hr respectively to obtain water extractive solution, concentrating the water extractive solution, and drying to obtain water extract; uniformly mixing 10g of rosemary volatile oil and 5g of rose volatile oil, adding beta-cyclodextrin which is 5 times of the total weight of the volatile oil for clathration, drying at 55-60 ℃, and crushing to obtain a volatile oil clathrate compound; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition.
Example 6
500g of lactose was added to 500g of the pharmaceutical composition prepared in example 1, and the mixture was uniformly mixed and granulated by dry compression to obtain granules.
Example 7
Adding 70g of starch into 260g of the pharmaceutical composition prepared in the method of example 2, uniformly mixing, granulating by using 85% ethanol as a wetting agent, drying at the temperature of below 60 ℃, grading, and filling into hard capsules to obtain hard capsules.
Example 8
400g of the pharmaceutical composition prepared by the method of example 3 is added with 200g of starch and 50g of sodium carboxymethyl starch, the mixture is uniformly mixed, 80% ethanol is used as a wetting agent, the mixture is granulated, dried and granulated, 30g of magnesium stearate is added, the mixture is uniformly mixed, and the mixture is tableted and coated with a film to obtain tablets.
Example 9
The pharmaceutical composition prepared by the method of example 4 is uniformly mixed with starch in a weight ratio of 1:1 to obtain powder.
Example 10
Adding 300g of soybean oil and 50g of beeswax into 100g of the pharmaceutical composition prepared in the example 5, heating and stirring uniformly, and pressing into soft capsules to obtain the soft capsules.
Comparative example 1
Taking 32g of ginseng, 80g of rhizoma acori graminei, 160g of poria cocos and 32g of polygala tenuifolia, and crushing into fine powder to obtain the medicinal composition.
Comparative example 2
Taking 320g of ginseng, 800g of rhizoma acori graminei and 320g of polygala tenuifolia, uniformly mixing, crushing into coarse powder, placing the coarse powder in an extraction kettle of a supercritical carbon dioxide extraction apparatus, extracting at the temperature of 50 ℃, under the extraction pressure of 28mPa for 90min, and obtaining mixed volatile oil and dregs; adding 5 times of beta-cyclodextrin into the mixed volatile oil for inclusion, drying at 55-60 ℃, and crushing to obtain a volatile oil inclusion compound; mixing the residue with 1600g Poria, decocting in water for 2 times (water amount per time is 24.32 kg) and each time for 1 hr to obtain water extractive solution, concentrating, drying, and pulverizing to obtain water extract; mixing the water extract and the volatile oil clathrate uniformly to obtain the pharmaceutical composition. The medicine composition and starch are uniformly mixed according to the weight ratio of 1:1 to obtain powder.
Examples of the experiments
The antidepressant effect of the pharmaceutical composition is verified by pharmacodynamic tests. The experimental method and results are as follows:
1. animals and materials
Kunming mouse, SPF grade, female, weight 20 + -2 g, provided by Experimental animals center of the fourth military medical university of people's liberation force of China.
Adrenocorticotropic hormone (ACTH) ELISA detection kit (Nanjing Biotechnology Co., Ltd.)
Electronic balance (Shanghai Minqiao precision scientific instruments Co., Ltd.)
Spevoramax 190 continuous spectrum microplate reader (America molecular instruments Co., Ltd.)
KDC-1044 Low-speed centrifuge (Anhui Zhongkejia scientific instruments Co., Ltd.)
DHP-9162 type electric constant temperature incubator (Shanghai-Hengshi Co., Ltd.)
DHG-9075A type electric heating constant temperature air-blast drying oven (Shanghai-Hengshi Co., Ltd.)
Haixin refrigerator-freezer BCD-301WT/A (Haixin group Co., Ltd.)
Control 1 group test solutions: and (3) adding distilled water into the pharmaceutical composition of the comparative example 1 to prepare a suspension containing 0.2g of crude drugs per 1ml, thus obtaining the pharmaceutical composition.
Control 2 test solutions: and (3) adding distilled water into the powder of the comparative example 2 to prepare a suspension containing 0.2g of the crude drug per 1ml, thus obtaining the traditional Chinese medicine composition.
Test solutions of experimental groups: the powder in example 9 is taken and added with distilled water to prepare a suspension containing 0.2g of crude drugs per 1ml, thus obtaining the traditional Chinese medicine.
2. Test method
2.1 mouse body weight and behavioural test
Taking 50 mice adapted to experimental environment, each half of the mice is female and male, selecting 10 mice as blank groups, and molding 40 mice. The weight of each mouse is respectively weighed and recorded one day before the experiment begins, the mice are placed into a 50ml centrifuge tube every day after the model building begins, small holes with the diameter of 0.4cm are chiseled on the tube wall to ensure good ventilation, and lining cloth is placed at the bottom of a binding tube so as to absorb the excrement of animals. The mouse moves in a limited space in the restraint tube, but the injury on the body can not be caused, the mouse is placed into a water bath at room temperature to be soaked to the xiphoid process of the mouse in a water bath manner, the water is restrained for 1 hour every day, and the molding is continuously carried out for 7 days. The model mice were divided into model group, comparative 1 group, comparative 2 group and test group, 10 mice per group. Mice in blank group and model group were gavaged with 5 mL/kg per day－1The distilled water of (1); the control group 1 and the control group 2 are administered with the test solution of the control group 1 and the test solution of the control group 2 respectively at a dose of 1.0 g/kg per day-1(ii) a Test group the test solution of the present invention was administered to the test group at a dose of 1.0 g/kg/day-1. Except for the blank group, the model group, the control 1 group, the control 2 group and the experimental group were administered daily while continuing to be immersed in the restraining water for 1 hour for two consecutive weeks, and the body weight of each mouse was weighed once every 7 days to calculate the rate of change of body weight. The open field test and forced swimming test were performed two weeks later.
2.2 mouse sweet water preference degree experimental method
After the mice are adaptively raised, 1% sucrose intake training is carried out, namely, the mice are fed with 1% sucrose water and drinking water for 48 hours at the same time, then the water is forbidden for 24 hours, the mice are freely drunk with 1% sucrose water and drinking water for 1 hour in the next morning (taken as the 0 th day) at the same time, and the intake quantities of the mice are respectively recorded. The 1% sucrose water intake and the drinking water intake in 1h were measured at 8: 00-9: 00 in the early morning of 1 st, 2 nd, 3 rd weekends of the experiment, respectively.
3. Observation index
3.1 weight detection:
the weight of each mouse was weighed once every 7 days and the change in weight of the mice was recorded.
3.2 open field test
Changes in the behaviour of the groups of mice were studied in the open field trial at 1d and 22d before the trial. The mouse is placed in the center of an open field test box, the number of the grids which the mouse runs through from the center of the bottom surface within 5min, the number of times of rear leg standing (including counterfort standing and free standing) and the number of times of hair removal are recorded, and the test is carried out in a dark room and a quiet environment.
3.3 forced swimming test
Forced swim tests were performed on day 22. Placing a single mouse into an organic glass cylinder with the diameter of 18cm and the height of 40cm, and placing the cylinder in a sound-insulation dark box, wherein the water depth in the cylinder is 20cm, and the water temperature is room temperature. The activity of the mouse is collected and recorded through the camera, the swimming condition of the mouse within 5min is observed, and the immobility time of the mouse within 5min is counted. The state in which the head is merely kept on the water surface without being flooded by floating is judged as the swimming indeterminate state. The cumulative percent motionless time is calculated.
3.4 measurement of plasma adrenocorticotropic hormone (ACTH)
After the open field test and the forced swimming test are finished, blood is taken out, placed into an anticoagulation tube containing EDTA, kept stand for 2h at the temperature of 4 ℃, centrifuged at 3000r/min for 15min, the upstream blood plasma is taken out and sucked out by a pipette tip and placed into a test tube, stored at the temperature of minus 20 ℃, and the blood plasma adrenocorticotropic hormone level is measured by an enzyme-linked immunosorbent assay.
3.5 measurement of consumption of sugar Water and pure Water
1% sucrose water intake ═ 1% sucrose water intake/(1% sucrose water intake + drinking water intake) ] × 100%.
4. Test results
4.1 mouse weight
As can be seen from Table 1, the body weights of the mice of each group on day 0 and day 7 were not significantly different, but the body weights of the animals of each group molded by day 7 were significantly reduced; on the 21 st day of model building, compared with the model group, the body weight of the mice in the experimental group and the control group are obviously increased, and have significant difference (P is less than 0.05), and the body weight increase of the experimental group is higher than that of the control group.
TABLE 1 mouse weight Change results
| Group of | Example number (only) | Day 0 (g) | Day 7 (g) | Day 14 (g) | Day 21 (g) |
| Blank group | 10 | 20.09±1.28 | 22.13±1.14 | 24.10±1.12 | 25.52±1.23 |
| Model set | 10 | 20.17±1.21 | 21.30±1.26 | 22.39±1.14* | 23.41±1.30* |
| Control 1 group | 10 | 19.83±1.15 | 21.23±1.31 | 22.97±1.20 | 24.00±1.19△ |
| Control 2 group | 10 | 19.93±1.02 | 21.35±1.15 | 23.15±1.16 | 24.12±1.22△ |
| Experimental group | 10 | 20.01±1.09 | 21.55±1.21 | 23.39±1.25 | 24.41±1.14△ |
Note: in comparison with the blank set, the results,*p is less than 0.05; in comparison with the set of models,△P＜0.05。
4.2 open field test
As can be seen from Table 2, compared with the blank group, the number of crossing the grid, the number of times of standing and the number of times of hair trimming of the mice in the model group are obviously reduced, and the difference has statistical significance (P is less than 0.05); compared with the model group, the mice of the experimental group and the control group have obviously increased horizontal crossing grid number, standing times and hair trimming times, and the difference has statistical significance (P is less than 0.05). The number of horizontal crossing lattices, the standing times and the hair-managing times of the experimental group are increased compared with those of the control group, which shows that the invention is superior to the control group in the aspect of improving the depression degree of the model mouse.
TABLE 2 comparison of scores from mouse open field trials
Note: in comparison with the blank set, the results,*p is less than 0.05; in comparison with the set of models,△P＜0.05。
4.3 forced swim test
As can be seen from Table 3, the percentage difference in cumulative dead time for the model group compared to the blank group is statistically significant (P < 0.01); compared with the model group, the percentage of the dead time of the forced swimming in the experimental group has significant difference (P is less than 0.01), and the percentage of the dead time of the forced swimming in the control group has significant difference (P is less than 0.05), which shows that the invention is superior to the comparative example in the aspect of improving the depression degree of the model mouse.
TABLE 3 cumulative percent immobility time for mice
| Group of | Example number (only) | Cumulative percent of immobility time |
| Blank group | 10 | 37.12±7.34 |
| Model set | 10 | 65.46±6.15** |
| Control 1 group | 10 | 55.35±6.53△ |
| Control 2 group | 10 | 54.11±6.76△ |
| Experimental group | 10 | 48.31±8.32△△ |
Note: in comparison with the blank set, the results,**p is less than 0.01; in comparison with the set of models,△p is less than 0.05; in comparison with the set of models,△△P＜0.01。
4.4 measurement of plasma adrenocorticotropic hormone (ACTH)
As can be seen from Table 4, the plasma ACTH concentration in the model group was significantly increased compared to the blank group, and the difference was statistically significant (P < 0.01); compared with a model group, the concentration of ACTH in plasma of an experimental group is obviously increased and has obvious difference (P is less than 0.01), the concentration of ACTH in plasma of a control group is obviously increased and has obvious difference (P is less than 0.05), and the invention is superior to a comparative example in the aspect of improving the depression degree of a model mouse.
TABLE 4 comparison of plasma ACTH concentrations in mice
| Group of | Example number (only) | ACTH(pg/ml) |
| Blank group | 10 | 28.35±5.24 |
| Model set | 10 | 44.45±6.36** |
| Control 1 group | 10 | 36.12±3.87△ |
| Control 2 group | 10 | 34.42±3.43△ |
| Experimental group | 10 | 31.24±3.37△△ |
Note: in comparison with the blank set, the results,**p is less than 0.01; in comparison with the set of models,△p is less than 0.05; in comparison with the set of models,△△P＜0.01。
4.5 sucrose Water preference
As can be seen from Table 5, the sucrose water preference degrees of the mice of each group before molding have no significant difference; the intake of the model group mice on the 14 th day and the 21 st day of the model is obviously reduced compared with that before the model is made (P is less than 0.05); compared with the model group, the intake of the experimental group and the control group of mice is obviously increased, and the difference has statistical significance (P is less than 0.05). The intake of sucrose water in the experimental group was greater than that in the control group.
TABLE 5 comparison of sucrose preferences in mice
Note: in comparison with the blank set, the results,*p is less than 0.05; in comparison with the set of models,△P＜0.05。
in conclusion, the traditional Chinese medicine composition can increase the body quality of a depression model mouse, improve the sucrose water intake of the model mouse and obviously improve the plasma ACTH concentration of the model mouse; the open field test ethology shows that the traditional Chinese medicine composition can obviously increase the crossing number, standing times and hair trimming times of the mice in the model group; forced swimming experiments show that the invention can obviously increase the percentage of the immobility time of the model mouse. The invention can obviously improve the depression degree of a model mouse and has good antidepressant effect.
The present invention is not limited to the above-described embodiments, and any variations, modifications, and substitutions which may occur to those skilled in the art may be made without departing from the spirit of the invention.
Claims (2)
1. A preparation method of a medicinal composition for resisting depression is characterized by comprising the following steps of:
(1') extracting volatile oil from ginseng, rhizoma acori graminei and polygala tenuifolia to obtain mixed volatile oil and dregs;
(2') clathrating the mixed volatile oil, the rosemary volatile oil and the rose volatile oil with cyclodextrin to obtain a volatile oil clathrate;
(3') extracting the residue and Poria cocos wolf with water as solvent to obtain water extract, concentrating the water extract, and drying to obtain water extract;
(4') uniformly mixing the volatile oil inclusion compound and the water extract to obtain the pharmaceutical composition;
wherein, 20 to 60 parts by weight of ginseng, 40 to 100 parts by weight of acorus gramineus, 60 to 200 parts by weight of poria cocos, 18 to 40 parts by weight of polygala tenuifolia, 1 to 2 parts by weight of rosemary volatile oil and 0.5 to 1 part by weight of rose volatile oil;
wherein the content of the first and second substances,
in the step (1'), the method for extracting the volatile oil is a steam distillation method, and comprises the following steps: adding water into the ginseng, the acorus gramineus soland and the polygala tenuifolia for steam distillation, wherein the water adding amount is 8-15 times of the total weight of the ginseng, the acorus gramineus soland and the polygala tenuifolia; distilling for 4-7 hours to obtain mixed volatile oil and an extracting solution; filtering the extractive solution to obtain medicinal liquid and residue; or
In the step (1'), the method for extracting the volatile oil is a supercritical extraction method, and comprises the following steps: extracting the ginseng, the rhizoma acori graminei and the polygala tenuifolia into volatile oil by adopting a supercritical carbon dioxide extraction method, wherein the extraction temperature is 45-65 ℃; the extraction pressure is 25-32 MPa; extracting for 60-120 min to obtain the mixed volatile oil;
in the step (2'), uniformly mixing the mixed volatile oil, the rosemary volatile oil and the rose volatile oil, and then performing inclusion by using beta-cyclodextrin to obtain a volatile oil inclusion compound; the dosage of the beta-cyclodextrin is 2-6 times of the total weight of the mixed volatile oil, the rosemary volatile oil and the rose volatile oil;
in the step (3'), the extraction method is a decoction method or a reflux extraction method; the preparation method of the water extracting solution comprises the following steps: and mixing the decoction dregs and the poria cocos, adding water for extraction for 1-4 times, wherein the water amount of each time is 5-15 times of the total weight of the ginseng, the rhizoma acori graminei, the polygala tenuifolia and the poria cocos, and the extraction time of each time is 0.5-3 hours, so that the water extract is obtained.
2. The method according to claim 1, wherein,
20 parts of ginseng, 100 parts of rhizoma acori graminei, 200 parts of poria cocos, 20 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
60 parts of ginseng, 40 parts of rhizoma acori graminei, 60 parts of poria cocos, 40 parts of polygala tenuifolia, 1 part of rosemary volatile oil and 0.5 part of rose volatile oil; or
32 parts of ginseng, 40 parts of rhizoma acori graminei, 160 parts of poria cocos, 40 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
32 parts of ginseng, 80 parts of acorus gramineus, 160 parts of poria cocos, 32 parts of polygala tenuifolia, 2 parts of rosemary volatile oil and 1 part of rose volatile oil; or
20 parts of ginseng, 40 parts of acorus gramineus, 70 parts of poria cocos, 18 parts of polygala tenuifolia, 1 part of rosemary volatile oil and 0.5 part of rose volatile oil.
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