CN115998793B - Vigour-preserving soup for improving learning and memory capacity of organism and preparation method thereof - Google Patents
Vigour-preserving soup for improving learning and memory capacity of organism and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a health care soup for improving learning and memory ability of an organism, which comprises the following components: the invention also relates to a preparation method of the health care soup for improving the learning and memory capacity of organisms. The beneficial effects of the invention are as follows: the invention obtains the primordial energy-preserving soup of the three formulas through the integral regulation effect of multiple links and multiple targets, has remarkable effect on improving the learning and memory capacity of the memory-impaired mice, and has the protection effect on the nerves of the memory-impaired mice; meanwhile, the invention researches in vivo experiments, and the in vivo experiments prove that the Baoyuan decoction for improving the learning and memory ability of the organism can restore and protect the learning and memory ability of mice with Alzheimer's disease, and can be used as a potential treatment means of Alzheimer's disease; the invention can be applied to the preparation of medicaments or health care products for improving the learning and memory capacity of organisms.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a primordial energy-preserving soup for improving learning and memory capacity of an organism and a preparation method thereof.
Background
With the acceleration of social development, the phenomenon of aging of population is gradually prominent. Alzheimer's disease is the most common neurodegenerative disease which is easy to get for middle-aged and elderly people, and brings serious physical and psychological burden to patients and families thereof. In order to ensure the later life happiness of the old, improve the resident happiness life index and improve the learning and memory capacity of the middle-aged and the old, the method is probably a method for preventing the Alzheimer disease.
Through research, it has now been found that Baoyuan decoction has neuroprotective effects of resisting oxidation, inhibiting inflammation and resisting apoptosis. However, whether the Baoyuan decoction can improve the learning and memory ability is reported recently.
Molding the mice by injecting streptozotocin into a single ventricle; the injection of streptozotocin into the ventricles of the mice can cause the energy metabolism in the brains of the mice to be disturbed, reduce the energy supply level in the brains, simultaneously cause related oxidative damage to brain areas of the mice, induce the expression of inflammatory factors to rise, promote the occurrence of neuroinflammation, finally promote the decline of learning and memory abilities of the mice and the change of behaviours. The model simulates sporadic Alzheimer's disease, and is suitable for research of Alzheimer's disease on learning and memory ability injury.
Disclosure of Invention
The invention aims to overcome the defects of the existing medicines capable of treating Alzheimer disease, and provides a health care soup which has no obvious side effect on human bodies, can be used for a long time and improves the learning and memory capacity of organisms and a preparation method thereof.
The first Baoyuan decoction for improving the learning and memory capacity of the organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice and 1.77g of cinnamon.
The second type of Baoyuan decoction for improving the learning and memory capacity of the organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice, 1.77g of cinnamon and 8.85g of shizandra berry.
The third kind of Baoyuan decoction for improving the learning and memory ability of the organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice, 1.77g of cinnamon, 13.275g of cassia twig, 8.85g of bighead atractylodes rhizome, 26.55g of poria cocos and 17.7g of angelica.
The preparation method of the three primordial qi-preserving soup for improving the learning and memory capacity of the organism comprises the following specific steps:
Selecting traditional Chinese medicine raw materials according to a set weight ratio, adding ginger, and soaking with water for a set period of time;
Adding water, decocting for the first time, and filtering with gauze;
Adding water into the decoction obtained by filtering with gauze after the first decoction, decocting for the second time, and filtering with gauze to obtain the health care soup for improving learning and memory ability of organism.
Preferably, the traditional Chinese medicine raw materials are selected according to the set weight ratio, and then are soaked in water for 30min after being added with a piece of ginger.
Preferably, 400ml of water is added in the first decoction, the decoction is carried out for 90min, and the mixture is filtered by using 200-mesh gauze.
Preferably, in the second decoction, 100ml of water is added into the decoction obtained by filtering the gauze after the first decoction, and the decoction is decocted for 30min and is continuously filtered by using 200-mesh gauze.
The three applications of the vitality-preserving soup for improving the learning and memory capacity of the organism are used for preparing medicaments for improving the learning and memory capacity of the organism.
The beneficial effects of the invention are as follows:
the invention provides the health care soup which has no obvious side effect on human bodies, can be used for a long time, improves the learning and memory capacity of organisms, utilizes the advantages of multiple targets and multiple links of traditional Chinese medicines through a controlled variable method, comprehensively compares the treatment effects of the health care soup prepared from traditional Chinese medicine raw medicinal materials with different components and dosages, and obtains the optimal dosages of various traditional Chinese medicine raw medicinal materials in the health care soup;
The invention obtains the primordial energy-preserving soup of the three formulas through the integral regulation effect of multiple links and multiple targets, has remarkable effect on improving the learning and memory capacity of the memory-impaired mice, and has the protection effect on the nerves of the memory-impaired mice; meanwhile, the invention researches in vivo experiments, and the in vivo experiments prove that the Baoyuan decoction for improving the learning and memory ability of the organism can restore and protect the learning and memory ability of mice with Alzheimer's disease, and can be used as a potential treatment means of Alzheimer's disease; the invention can be applied to the preparation of the medicine for improving the learning and memory ability of organisms.
Drawings
FIG. 1 is a graph comparing changes in the fourth quadrant of the water maze mice for five days of incubation;
FIG. 2 is a histogram of changes in latency of groups of mice versus Model group on day five of the fourth quadrant of the water maze;
FIG. 3 is a histogram of the proportion of stiff time to total time for each group of mice in a conditional fear experiment;
FIG. 4 is a histogram of total cholesterol content in serum of each group of mice versus Model group;
FIG. 5 is a histogram of the comparison of the high density lipoprotein content in the serum of each group of mice with the Model group;
FIG. 6 is a histogram of the comparison of the low density lipoprotein level in the serum of each group of mice with the Model group;
FIG. 7 is a histogram of triglyceride levels in serum of mice of each group versus Model group;
FIG. 8 is a schematic representation of representative bands of related proteins in the hippocampus of each group of mice;
FIG. 9 is a histogram showing the comparison of SHANK3 protein expression in hippocampus of mice in each group with Model group;
FIG. 10 is a histogram of NR2A expression in hippocampus vs. Model group for each group of mice;
FIG. 11 is a histogram showing the comparison of PSD95 protein expression in hippocampus of mice in each group with Model group;
FIG. 12 is a histogram of NR2B protein expression in hippocampus vs. Model group for each group of mice.
Detailed Description
The invention is further described below with reference to examples. The following examples are presented only to aid in the understanding of the invention. It should be noted that it will be apparent to those skilled in the art that modifications can be made to the present invention without departing from the principles of the invention, and such modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.
Example 1
A Baoyuan decoction for improving the learning and memory capacity of an organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice and 1.77g of cinnamon.
Example 2
A Baoyuan decoction for improving the learning and memory capacity of an organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice, 1.77g of cinnamon and 8.85g of shizandra berry.
Example 3
A Baoyuan decoction for improving the learning and memory capacity of an organism is prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice, 1.77g of cinnamon, 13.275g of cassia twig, 8.85g of bighead atractylodes rhizome, 26.55g of poria cocos and 17.7g of angelica.
Example 4
Based on the embodiment 1, 2 or 3, the embodiment 4 of the application provides a preparation method of the Baoyuan decoction for improving the learning and memory ability of organisms:
Taking several traditional Chinese medicine raw materials of ginseng, astragalus, liquorice, cinnamon, schisandra chinensis, cassia twig, bighead atractylodes rhizome, poria cocos and angelica according to the formula of the embodiment 1, 2 or 3, adding a piece of ginger, and soaking for 30min;
performing primary decoction: adding 400ml of water, decocting for 90min, and filtering with 200 mesh gauze;
and (3) performing secondary decoction: adding 100ml of water into the decoction obtained by filtering gauze after the first decoction, and decocting for 30min; filtering with 200 mesh gauze to obtain the health soup for improving learning and memory ability.
Example 5
Based on examples 1 to 4, the present example uses three primordial energy-preserving soup for improving learning and memory ability of organism prepared by the formulation of example 4 according to example 1 to example 3 in learning and memory ability experiment of single-sided ventricle injection streptozotocin mice to detect the effect of primordial energy-preserving soup for improving learning and memory ability of organism on learning and memory ability of Alzheimer disease mice:
Experimental materials:
1. Experimental biology: experimental animals: strain ICR mice (SPF grade);
2. medicine: streptozotocin, available from Sigma, usa under the product number V900890, purity 98%;
3. Seven dosing groups were set: the group B1 is administered by using the primordial energy preserving soup of the corresponding formula in the embodiment 1, the group B2 is administered by using the primordial energy preserving soup of the corresponding formula in the embodiment 2, and the group B3 is administered by using the primordial energy preserving soup of the corresponding formula in the embodiment 3; the ginseng, astragalus root, licorice, cinnamon, schisandra fruit, cassia twig, bighead atractylodes rhizome, poria cocos and angelica in the Baoyuan decoction components in the B1 group to the B3 group are purchased from Beijing Tongren Tang; the control group is a normal control group, the sham group is a sham operation group (i.e. a group which performs related operations but does not use related medicines), the model group is a model group, and the done group is a positive medicine group;
4. experimental instrument and materials
The experimental process comprises the following steps:
5.1, modeling and drug delivery modes:
An amount of STZ powder was weighed and dissolved in 0.01m μl-1PBS at ph=4.5 to prepare a50 μg μl-1 solution. Mixing well, wrapping the EP pipe with tinfoil paper, and avoiding light transmission. The molding was performed by lateral ventricle STZ injection, mice were anesthetized with 0.3% sodium pentobarbital solution (50 mg kg-1) and subsequently fixed, and head hair was cut off to expose scalp. The scalp is cut off, the lower part of the anterior cranium is 0.5mm, the horizontal side is 1.0mm, and the needle insertion depth is 3.0mm. The STZ groups each injected 3 μl and the sham groups each injected an equal volume of PBS. After molding, mice were sutured, adaptively bred for one week, then randomly grouped, and dosed.
5.2, Spatial learning and memory experiment water maze experiment:
After the end of the administration, the mice were subjected to a water maze experiment for five days; the first 4 days are training periods, each mouse is placed in water in four quadrants in sequence, each quadrant is provided with a graph with different shapes as a mark, the mouse needs to face the graph when placed on the water surface, the recording time is 60s, and if the mouse cannot find a platform within 60s, the mouse is guided to the platform. Mice were tested on day 5 for escape latency, and data were collected and processed by the water maze image automatic monitoring system to obtain a graph comparing the change profiles of the four-quadrant five-day latency of each group of mice in the water maze as shown in fig. 1.
5.3, Conditional fear experiments:
For three days. Placing the mice in a dark cage on the first day, adapting the mice for 3min, then starting acoustic stimulation (Tone, 2.5hz,85 db) for 30 seconds, performing electric shock (0.75 mA) on the soles of the mice after the acoustic stimulation is finished, and silencing for 2min after the electric shock is finished; the above acoustic stimulus-electrical stimulus-quiet period procedure was cycled a total of 7 times, with a total duration of 1244s. The next day, mice were placed in the same cages and allowed to freely move without any stimulation for 1200s. The third day a memory test was performed.
5.4, Behavioural experiments:
Changing the internal environment of the cage, covering the original inner wall of the cage with a paperboard, opening light, putting the mice into the cage, adapting to the light for 3min, opening acoustic stimulus (Tone, 2.5hz,85 db) for 30s, and after finishing, silencing for 2min, repeating the process for 10 times in total, wherein the whole experiment needs to ensure that the external environment is quiet and light-proof.
5.5, Detection of triglycerides and total cholesterol in serum:
After the behavioural experiment is completed, the mice are anesthetized, sacrificed by cervical removal, brain tissues are separated, the hippocampus is rapidly peeled off on ice, and the frozen products are placed in a minus eighty degree refrigerator for freezing. The detection of triglycerides and total cholesterol was performed against serum according to the kit instructions.
Taking out Hippocampus in minus eighty degrees refrigerator, adding RAPA lysate, PMSF, and phosphatase inhibitor, cutting, performing tissue disruption in an ultrasonic disruption device, centrifuging at 4deg.C in 12000g min-1 centrifuge for 15min, collecting supernatant, detecting sample concentration with BCA kit, and adding loading buffer at 95deg.C for 8min. SDS-PAGE electrophoresis for 45min, NC membrane wet-transfer for 65min, then placing the sheared strips in 5% milk prepared with TBST buffer for one hour at room temperature, placing the strips in primary antibodies (NR 2B, NR2A, shank antibodies diluted 1:1000; PSD95 diluted 1:4000) after one hour, and cooling overnight at 4 ℃. After gradient washing, secondary antibody (dilution ratio 1:10000) is added for incubation for one hour at room temperature, ECL luminous solution is soaked after gradient elution, a Tanon 5200Multi gel imaging system scans strips, and Image J software is used for analysis.
The statistical method adopts single-factor analysis of variance and GRAPHPAD PRISM software to analyze.
Results
The change in latency of mice in each group on day five of the fourth quadrant of the water maze was compared to the Model group (n=4), a: p < 0.05, giving the results as in fig. 2: the dose of the Baoyuan decoction and the prescription of the Baoyuan decoction in the embodiment 1 to the embodiment 3 is 2 times that of the Baoyuan decoction, so that the dead time ratio in the space fear experiment of the mice injected with streptozotocin by the lateral ventricle can be increased; as shown in fig. 3, the proportion of the stiff time of each group of mice in the fear test is compared with the total time, and the conclusion that the primordial care soup and the 2-time dosage added by the primordial care soup in the examples 1 to 3 can improve the blood index of the mice injected with streptozotocin from the lateral ventricle is obtained; as shown in FIG. 4, the total cholesterol content in the serum of each group of mice was compared with that of the Model group, a: p < 0.05, c: p < 0.001; as shown in FIG. 5, the serum content of high density lipoprotein in each group of mice was compared with that of the Model group, a: p < 0.05, b: p < 0.01; as shown in FIG. 6, the serum low density lipoprotein content of each group of mice was compared with that of the Model group, b: p < 0.01, c: p < 0.001; as shown in FIG. 7, the triglyceride content in the serum of each group of mice was compared with that of the Model group, a: p < 0.05, b: p < 0.01; as shown in fig. 8, 2-fold doses of the group's primordial energy-preserving soup and its formulation promote expression of synapse-related proteins in mice injected with streptozotocin from the ventricles; as shown in fig. 9 to 12, the expression of the SHANK3 protein (a: p < 0.05), the expression of NR2A (a: p < 0.05, B: p < 0.01), the expression of PSD95 protein (a: p < 0.05) and the expression of NR2B protein ((n=4), a: p < 0.05, B: p < 0.01) in the hippocampus of each group of mice were compared with the Model group (n=4), respectively; in the above, n is the sample content, a, b and c are the corresponding statistical p-value, wherein the p-value corresponding to a is smaller than 0.05, the p-value corresponding to b is smaller than 0.01, and the p-value corresponding to c is smaller than 0.001;
taken together, the 2-fold doses of the Baoyuan decoction and the Baoyuan decoction in the embodiment 1 to the embodiment 3 can generate better memory protection effect on mice injected with streptozotocin into the lateral ventricle. The duration of the fifth day water maze incubation period of the mice can be reduced. Increasing the duty cycle of the dead time in the conditioned fear experiments. Increase expression of related synaptoproteins in the hippocampus of mice.
Claims (6)
1. The Baoyuan decoction for improving the learning and memory capacity of the organism is characterized by being prepared from the following traditional Chinese medicine raw materials in parts by weight: 8.85g of ginseng, 17.71g of astragalus root, 4.43g of liquorice, 1.77g of cinnamon, 13.275g of cassia twig, 8.85g of bighead atractylodes rhizome, 26.55g of poria cocos and 17.7g of angelica.
2. A method for preparing the health care soup for improving the learning and memory ability of an organism according to claim 1, which is characterized by comprising the following specific steps:
Selecting traditional Chinese medicine raw materials according to a set weight ratio, adding ginger, and soaking with water for a set period of time;
Adding water, decocting for the first time, and filtering with gauze;
Adding water into the decoction obtained by filtering with gauze after the first decoction, decocting for the second time, and filtering with gauze to obtain the health care soup for improving learning and memory ability of organism.
3. The method for preparing the health care soup for improving the learning and memory ability of the organism according to claim 2, which is characterized in that: selecting the traditional Chinese medicine raw materials according to the set weight ratio, adding a piece of ginger, and soaking in water for 30min.
4. The method for preparing the health care soup for improving the learning and memory ability of the organism according to claim 2, which is characterized in that: adding 400ml of water in the first decoction, decocting for 90min, and filtering with 200 mesh gauze.
5. The method for preparing the health care soup for improving the learning and memory ability of the organism according to claim 2, which is characterized in that: in the second decoction, 100ml of water is added into the decoction obtained by filtering the gauze after the first decoction, and the decoction is decocted for 30min and is continuously filtered by using 200 meshes of gauze.
6. Use of the baoyuan decoction for improving learning and memory ability of an organism according to claim 1 in preparation of a medicament for improving learning and memory ability of an organism.
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