CN110237204B - A composition with brain strengthening/cranial nerve regulating effects - Google Patents

A composition with brain strengthening/cranial nerve regulating effects Download PDF

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CN110237204B
CN110237204B CN201910648608.8A CN201910648608A CN110237204B CN 110237204 B CN110237204 B CN 110237204B CN 201910648608 A CN201910648608 A CN 201910648608A CN 110237204 B CN110237204 B CN 110237204B
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parts
composition
brain
strengthening
effects
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CN110237204A (en
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姜海燕
易斌
罗丽莲
肖艳皎
韩雅慧
陈明霞
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Beijing Silian Pharmaceutical Industry Co ltd
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Beijing Silian Pharmaceutical Industry Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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Abstract

The invention discloses a composition with brain strengthening/cranial nerve conditioning effects, which comprises the following components in parts by weight: 0.0002 to 0.0005 part of folic acid compound, 2 to 4.5 parts of ginkgo leaf, 0.5 to 1.5 parts of galangal, 1 to 3 parts of white paeony root, 0.5 to 1.5 parts of szechuan lovage rhizome, 3 to 5 parts of danshen root, 1 to 3 parts of common anemarrhena rhizome and 1 to 3 parts of barbary wolfberry fruit. Compared with the prior art, the composition has the effects of improving sleep and enhancing memory, and has good effects of nourishing cranial nerves, strengthening brain and improving intelligence.

Description

A composition with brain strengthening/cranial nerve regulating effects
Technical Field
The invention relates to a composition, in particular to a composition with brain strengthening/cranial nerve conditioning effects.
Background
The brain, functioning as a high-level center regulating various systems and organs of the body, participates in and regulates neural activities such as learning, memory, language, and consciousness. The brain function is closely related to human life, and the abnormal brain function affects the cognition, emotion and behavior of human and the function of each organ to different degrees, further affecting the daily life, work and study, social activities and the like of human. However, with the increasing frequency of social and economic activities and the increasing pace of modern life, people in all levels of society have more and more obviously realized the heavy burden of mental stress to people in recent years. More and more people, in a sub-healthy state of the nerves due to insufficient nutrition and excessive stress on the brain, stand out as: mental fatigue, inattention, physical weakness, memory loss, comprehension and creativity loss, dysphoria, anxiety, and immune dysfunction. Therefore, there is an increasing demand for nutritional health products having brain strengthening and cranial nerve regulating effects.
CN108497493A discloses a food for improving the function of cranial nerve cells, which comprises the following raw materials in parts by weight: 50-200 parts of emblic leafflower fruit extract, 5-50 parts of phosphatidylserine, 5-50 parts of sodium hyaluronate, 1.5-15 parts of gamma-aminobutyric acid, 0.3-3 parts of calcium pantothenate and 6-30 parts of epsilon-polylysine hydrochloride. The food for improving brain nerve cell function has effects of regulating nerve impulse conduction, improving brain memory function and repairing damaged brain cell.
CN108714192A discloses a composition for improving the function of cerebral neurotrophic factors, which comprises, by weight, 91.9-99.8% of asparagus extract, 0.3-2.4% of phosphatidylserine, 0.3-2.4% of fungus extract, 0.09-1.6% of theanine and 0.09-2.0% of gamma-aminobutyric acid.
With the development of society, the proportion of brainworkers is larger and larger, and with the development of mobile internet, the modern people generally have excessive brain use, and the population suffering from insomnia is larger and larger. Insomnia causes the brain to have insufficient rest for a long time, increases the burden of the brain, and can also cause some chronic diseases. Accordingly, there is an increasing demand for drugs and health products for treating and improving insomnia.
CN109512910A discloses a medicine for treating insomnia, which comprises the following components in parts by weight: 15 parts of radix bupleuri, 15 parts of fructus aurantii, 15 parts of red peony root, 6 parts of liquorice, 20 parts of salvia miltiorrhiza, 20 parts of radix achyranthis bidentatae, 20 parts of uncaria, 10 parts of chrysanthemum, 20 parts of rhizoma alismatis, 20 parts of ligusticum chuanxiong hort, 10 parts of rhizoma corydalis, 10 parts of polygala tenuifolia and 20 parts of albizia flower. CN109303841A discloses a traditional Chinese medicine composition for treating insomnia, which comprises the following raw materials in parts by weight: 10-25 parts of longan pulp, 15-20 parts of red date, 15-25 parts of lotus seed, 5-10 parts of peach seed, 15-25 parts of hawthorn, 10-15 parts of cassia seed and 8-10 parts of lily. The medicine has functions of tranquilizing mind, refreshing brain, and treating insomnia. CN104306555A discloses a food, health care product or pharmaceutical composition for treating insomnia, which comprises the following raw materials in parts by weight: 8-15 parts of coreopsis tinctoria, 5-15 parts of lycium ruthenicum murr, 10-25 parts of lycium barbarum leaves, 12-15 parts of safflower and 15-25 parts of buckwheat. The invention carries out scientific and reasonable compatibility of the raw materials, exerts the synergistic interaction effect, can effectively improve the sleep condition and has good treatment and prevention effects on insomnia.
At present, in the prior art, aiming at the problems of single function and unobvious application effect of products for strengthening and regulating the cranial nerves, the development of a product for strengthening and regulating the cranial nerves with complete functions and good effect is urgently needed.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a composition with the brain strengthening/brain nerve conditioning effect, which not only has the effects of nourishing the brain nerves and strengthening the brain and improving intelligence, but also has the effects of improving sleep and enhancing memory.
In order to achieve the above object, the present invention provides the following technical solutions:
a composition with brain strengthening/cranial nerve conditioning effects comprises the following components in parts by weight:
Figure BDA0002134407070000021
preferably, the folic acid compounds are one or more of folic acid, leucovorin and corresponding salts thereof, L-5-methyltetrahydrofolic acid and corresponding salts thereof, pharmaceutically acceptable salts of folic acid, active metabolites of folic acid or pharmaceutically acceptable salts of folic acid, and substances capable of metabolizing and/or generating folic acid in vivo.
The technical effects of the invention can be realized by the above technical solutions, but in some preferred embodiments, the achieved technical effects are superior to other solutions.
For example:
when the mass ratio of the ginkgo leaves to the galangal to the rhizoma anemarrhenae is 3:1:2, one preferable composition comprises the following specific components in parts by weight: folic acid 0.00035 parts, ginkgo leaf 2.7 parts, galangal 0.9 parts, white peony root 2.5 parts, Ligusticum wallichii 1.2 parts, root of red rooted saliva 4.5 parts, anemarrhena rhizome 1.8 parts, wolfberry fruit 2.8 parts.
More preferably, the composition comprises the following specific components in parts by weight: 0.0004 part of folic acid, 3 parts of ginkgo leaf, 1 part of galangal, 2 parts of white paeony root, 1 part of szechuan lovage rhizome, 4 parts of salvia miltiorrhiza, 2 parts of rhizoma anemarrhenae and 2 parts of medlar.
In some preferred embodiments, the composition further comprises one or more of the following components in parts by weight:
Figure BDA0002134407070000031
most preferably, the composition comprises the following specific components in parts by weight: 0.0004 part of folic acid, 3 parts of ginkgo leaf, 1 part of galangal, 2 parts of white peony root, 1 part of szechuan lovage rhizome, 4 parts of salvia miltiorrhiza, 2 parts of rhizoma anemarrhenae, 2 parts of medlar, 4 parts of kudzuvine root, 3 parts of cassia seed, 3 parts of spina date seed, 1 part of sharpleaf galangal fruit, 2 parts of gardenia, 2 parts of perilla leaf, 1 part of platycladi seed, 5 parts of fiveleaf gynostemma herb, 1 part of ginseng, 2 parts of dodder, 1 part of Chinese magnoliavine fruit and 1 part of thinleaf milkwort root-bark.
The invention provides a preparation method of the composition with the brain strengthening/cranial nerve conditioning effect, which comprises the following steps:
(1) mixing the above compositions with brain nourishing/cranial nerve regulating effects except folic acid compounds, extracting with water for three times, mixing the filtrates, concentrating under reduced pressure, and drying to obtain Chinese medicinal extract;
(2) pulverizing the traditional Chinese medicine extract obtained in the step (1), sieving, and mixing with a formula amount of folic acid compound to obtain a composition with brain strengthening/cranial nerve regulating effects;
the invention also provides the application of the composition in preparing medicines and/or health products with brain strengthening/cranial nerve conditioning effects.
The invention also provides an oral preparation with brain strengthening/cranial nerve conditioning effects, which comprises the composition with the brain strengthening/cranial nerve conditioning effects and pharmaceutically acceptable auxiliary materials.
Preferably, the oral preparation is any one of tablets, capsules, granules, dripping pills, soft capsules and oral liquids.
Preferably, the auxiliary material includes any one of a disintegrant, a lubricant, and a binder.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a composition with brain strengthening/brain nerve conditioning effects, wherein folic acid compounds are compatible with ginkgo leaves, galangal, white paeony roots, ligusticum wallichii, salvia miltiorrhiza, rhizoma anemarrhenae, medlar and other traditional Chinese medicines, so that the synergistic interaction effect among the raw materials can be fully exerted, the effects of improving sleep and enhancing memory can be achieved, and the effects of better nourishing brain nerves and strengthening brain and improving intelligence can be achieved;
(2) by adding the traditional Chinese medicines such as the kudzuvine root, the cassia seed, the spina date seed, the sharpleaf galangal fruit, the gardenia, the perilla leaf, the platycladi seed, the gynostemma pentaphylla, the ginseng, the dodder, the schisandra chinensis, the polygala tenuifolia and the like into the composition, the memory is strengthened, and the effects of improving and enhancing the immunity of a human body, relieving depression and the like are achieved.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The raw materials used in the present invention are all common commercial products unless otherwise specified.
Examples 1 to 5
The raw material composition of a composition having brain strengthening/cranial nerve regulating effects of examples 1 to 5 is shown in table 1.
TABLE 1 raw material composition of a composition having brain strengthening/cranial nerve regulating effects
Figure BDA0002134407070000041
Figure BDA0002134407070000051
The method for preparing a composition having brain strengthening/cranial nerve regulating effects of examples 1 to 5, comprising the steps of:
(1) mixing the above raw materials except folic acid with water according to a material-to-liquid ratio of 1:6, extracting with water at 90 deg.C for three times (each time for 2 hr), mixing the three filtrates, concentrating under reduced pressure at 60 deg.C to relative density of 1.05, and drying under reduced pressure at 80 deg.C to obtain Chinese medicinal extract;
(2) pulverizing the traditional Chinese medicine extract obtained in the step (1) to 50 meshes, sieving, and mixing with folic acid with a formula amount to obtain the composition with the effects of strengthening brain and conditioning cranial nerves.
Examples 6 to 9
The raw material composition of a composition having brain strengthening/cranial nerve regulating effects of examples 6 to 9 is shown in table 2.
TABLE 2 composition of materials for a composition having brain strengthening/cranial nerve regulating effects
Raw materials Example 6 parts by weight Example 7 parts by weight Example 8 parts by weight Example 9 parts by weight
Folic acid 0.0004 0.0004 0.0004 0.0004
Ginkgo leaf 3 3 3 3
Galangal 1 1 1 1
White peony root 2 2 2 2
Rhizoma Ligustici Chuanxiong 1 1 1 1
Root of red-rooted salvia 4 4 4 4
Rhizoma anemarrhenae 2 2 2 2
Chinese wolfberry fruit 2 2 2 2
Kudzu root 3.5 5 4 4.5
Cassia seed 2 4.5 3 2.5
Wild jujube seed 2 4.5 3 4
Fructus alpiniae oxyphyllae 0.5 1.5 1 1.2
Gardenia jasminoides ellis 1 3 2 1.5
Folium Perillae 1.7 2.5 2 2.2
Arborvitae seed 0.5 1.5 1 0.8
Gynostemma pentaphylla 4 7.5 5 6
Ginseng radix 0.5 1.5 1 1.5
Semen Cuscutae 1 3 2 1.5
Schisandra chinensis 0.7 2 1 1.5
Root of Polygala 0.5 1.5 1 0.8
The method for preparing a composition having brain strengthening/cranial nerve regulating effects of examples 6 to 9, comprising the steps of:
(1) mixing the above materials except folic acid with water according to a material-to-liquid ratio of 1:6, extracting with water at 85 deg.C for three times (each time for 2 hr), mixing the three filtrates, concentrating under reduced pressure at 80 deg.C to relative density of 1.25, and drying under reduced pressure at 60 deg.C to obtain Chinese medicinal extract;
(2) pulverizing the traditional Chinese medicine extract obtained in the step (1) to 50 meshes, sieving, and mixing with folic acid with a formula amount to obtain the composition with the effects of strengthening brain and conditioning cranial nerves.
In order to better embody the advantages of the present invention, the animal experiments and results of the composition for invigorating brain/regulating cranial nerves provided by the present invention are given below.
Experimental example I, improving sleep
1. Experimental animals: SPF grade rats weighing 200-.
2. An experimental instrument: the sleep test box: the squirrel cage is formed by transforming a squirrel cage of 80cm multiplied by 40cm multiplied by 30cm, a small platform with the diameter of 6cm and a large platform with the diameter of 20cm are placed in the squirrel cage, a square plate filled with water is arranged at 1cm below the platform, and a feed box and a water bottle are arranged at the upper part of the platform; DW-5 type brain stereotaxic instrument, BL-420E biological function experiment system, Chengtai Union science and technology Limited company.
3. Experimental drugs: the compositions prepared in examples 1-9 of the present invention are diazepam, sodium pentobarbital and sodium penicillin for injection.
4. The experimental method comprises the following steps:
4.1 preparation of Insomnia rat model
The rats are respectively placed on small platforms in a sleep deprivation box, and the animals flex on the platforms to stand, freely take and drink water, can perform activities such as turning, raising heads, licking hairs and the like, and can doze off. This was repeated 7 times sleep deprivation, each for 24 hours, 1 time every other day, to achieve sleep deprivation effect. In order to eliminate the stress influence caused by isolation, activity limitation and water environment, a large platform control group is set, rats can sleep on the large platform without falling into water, and the large platform has the same diameter as the model group but can keep a complete sleep cycle because other environments are the same.
4.2 Experimental groups and dosing:
the weight groups were randomly divided into 10 groups, each group consisting of a large platform control group (normal group), a sleep deprivation model group (model group), a sleep deprivation model addition group (examples 1 to 9 of the present invention), and a sleep deprivation model addition stable group (western medicine control group). Rats were housed in cages for one week prior to the experiment and were fed and drunk freely.
Adding a medicine group: the compositions prepared in groups 1 to 9 of the present invention were administered by gavage every morning and every other day (no medication during sleep deprivation) at 10.0g/kg · bw. Grinding diazepam into fine powder, adding distilled water, shaking, and administering to the stomach at a dose of 0.5mg/kg every morning and once every other day (no drug is taken during sleep deprivation period). The model group and the normal group were drenched with the same volume of physiological saline, and the experiment was performed for 30 days.
4.3 Experimental indices
(1) General observations of animals: the general conditions of the animals, such as the overall shape (hair color, brightness and sparseness), the spirit (mental state, consciousness state and reaction sensitivity), the activity, the appetite, the weight change and the like, are observed every day.
(2) Polysomnography brain electrical activity: the sleep-wake cycle is divided into: wake period (W), slow wave sleep period (SWS, further divided into SWS1 and SWS2), and out-of-phase sleep Period (PS). Each recording was started at 9:00 am and was recorded 3 hours continuously, 3 times per rat, and the average was taken as a systematic arousal and sleep quality assessment.
The electrode establishing method comprises the following steps: SPF rats were anesthetized with pentobarbital sodium (40mg kg) by intraperitoneal injection-1Ip), fixing the head of a rat in a brain stereotaxic apparatus, cutting the skin on the top of the rat after disinfection, stripping the subcutaneous tissue, exposing the skull, cleaning the surface of the skull by hydrogen peroxide, and then respectively burying 2 clock small screws as brain electrodes at +/-2.0mm positions of the midline side from the front 2.0mm and the back 4.0mm without damaging dura mater. The muscle electrode and the eye electrode are made of enameled wires, wherein the muscle electrode is buried under deep neck muscles, the eye electrode is buried under one lateral eye outer canthus of a rat, 8 ten thousand doses of penicillin sodium are injected into abdominal cavity for 3 days immediately after operation, and the test is carried out after recovery for one week, so that the test record can be obtained.
4.4 results of the experiment
(1) General condition observation of experimental animals
The fatigue and the drowsiness of the model group and the western medicine control group are aggravated along with the increase of the sleep deprivation times, the most obvious symptoms are low mental state and reduced activity, the weight increment during the experiment is obviously lower than that of the normal group (P is less than 0.01), the mental state and the activity state of each group of examples 1-9 are better in performance, the fatigue degree is lighter, the fatigue state of the same degree is late, the weight increment is higher than that of the model group, wherein, the difference of the groups of examples 1-5 is obvious (P is less than 0.05) compared with the model group, and the difference of the groups of examples 6-9 is extremely obvious (P is less than 0.01) compared with the model group and has no obvious difference with the normal group. The results are shown in Table 3.
TABLE 3 general observations of experimental animals
Figure BDA0002134407070000071
Figure BDA0002134407070000081
Note: the ratio of # to normal group P is less than 0.05, and the ratio of # to normal group P is less than 0.01; the ratio P to model group was <0.05, P <0.01 to model group, as shown in the table below.
(2) Polysomnography brain electrical activity:
compared with the normal group, the PS of the model group is obviously shortened (P is less than 0.01) and the W is obviously prolonged (P is less than 0.05), which indicates that the insomnia model is successfully modeled; examples 1-5 each group significantly extended the SWS2 phase and the PS phase (P <0.05, P < 0.01); examples 6-9 showed a significant decrease in W phase (P <0.01), a significant increase in SWS2 phase (P <0.05) and PS phase (P <0.01) compared to the model group. The results of the groups of examples 6-9 are significantly better than the groups of examples 1-5, and are shown in Table 4.
TABLE 4 experimental animal polysomnography results/min
Group of W SWS1 SWS2 PS
Normal group 84.58±7.34 72.38±5.30 16.53±8.16 9.01±2.11
Model set 104.14±12.38# 60.58±10.34 14.00±7.14 3.93±2.19##
Example 1 88.69±11.08 68.17±4.53 17.28±8.16* 7.41±1.38*
Example 2 87.79±13.21 67.98±5.51 17.34±7.63* 7.38±1.21*
Example 3 86.81±13.19 71.68±8.43 17.52±9.18* 7.59±1.49*
Example 4 89.31±13.58 65.87±7.85 17.31±7.56* 7.24±1.10*
Example 5 84.73±12.33 72.28±8.14 17.81±8.89* 7.81±1.53*
Example 6 80.39±13.17* 77.71±8.97 18.68±9.71* 8.50±2.91**
Example 7 77.61±11.84* 75.68±9.63 19.16±10.83* 8.59±3.69**
Example 8 74.59±10.13* 78.35±7.50 19.68±12.80* 9.16±3.16**
Example 9 78.54±11.36* 76.28±8.14 19.57±11.39* 8.91±4.52**
Western medicine control group 81.32±13.45* 83.61±13.58 13.74±10.07 5.91±4.52
From the results, the traditional Chinese medicine composition has an obvious treatment effect on an insomnia animal model, can improve the overall state and the sleep time of an insomnia animal, has better performance in the activity state, the hair color, the food intake and the like of the insomnia animal compared with western medicine diazepam, and has the weight basically equivalent to that of a normal group during the whole experiment period.
Experiment example two: method for testing influence on learning and memory by diving platform method
1. Experimental animals: c57 white mouse, 6-8 weeks old, weight 20-24 g.
2. An experimental instrument: DT-200 diving tower automatic tester (Shanghai Yilian scientific and education Equipment Co., Ltd.).
3. Experimental dose: 6.7g/kg · bw.
4. The experimental method comprises the following steps:
4.1 Experimental groups and doses: randomly grouping mice according to body weight, setting 25 mice in each group, simultaneously setting a control group, and performing intragastric administration for 30 days according to the experimental dose, wherein the dosage is 0.2ml/10g · bw each time, and the control group is given with physiological saline.
4.2 diving platform experiment
The animals are placed in a reaction box to adapt for 3min, then 36V alternating current is conducted, the animals jump back to the insulating platform in normal reaction, most mice can jump back to the copper grid again or repeatedly, and the animals jump back to the platform rapidly after receiving electric shock. The training is started 2h after the gavage administration, the training is performed once every 3 days, and the formal test is performed 2h after the gavage administration on the 30 th day. The mice are placed on a platform in a reaction box, the first time of jumping of the mice (namely the electric shock latency) is recorded, the number of times of electric shock of the mice within 5min (the number of errors) is recorded, and the number is used as the learning achievement, and the specific data are shown in a table 5.
TABLE 5 mouse learning and memory test results
Latency period of electric shock/s Number of errors/time within 5min
Control group 162 3.6
Example 1 243 2.2
Example 2 248 2.0
Example 3 257 1.6
Example 4 246 2.1
Example 5 260 1.4
5. And (3) analyzing an experimental result:
table 5 shows the test data of the effect on learning and memory of mice, mainly the electric shock latency and the number of errors within 5min are recorded, and it can be seen from the experimental data that the compositions prepared in examples 1 to 5 of the present invention can prolong the electric shock latency of mice and significantly reduce the number of errors within 5min of mice compared with the control group. The effects of the embodiments 3 and 5 are obviously superior, and the results show that when the mass ratio of the ginkgo leaves to the galangal to the rhizoma anemarrhenae is 3:1:2, the memory of the mice can be obviously improved, and the effects of strengthening the brain are better.
Experimental example III Effect of composition on immunity
1. Experimental animals: clean grade ICR mice, male, weighing 18-22 g.
2. The experimental method comprises the following steps:
2.1 experimental grouping and dosing: the recommended amount of the composition for human body is 1.1 g/kg-bw per person per day, and the composition is taken after being mixed with warm water. Mice experimental dose settings were low, normal, and high at 3 dose levels, i.e., 5.5, 11, and 33g/kg · bw per day (corresponding to 5, 10, and 30 times the recommended amount for humans, respectively), while control groups were set, and experimental mice were randomly batched by body weight, 4 groups per batch, and 10 mice per group.
The stomach was gavaged once a day at a dose of 0.2ml/10g · bw, and the control group was given an equal volume of physiological saline for 30 consecutive days.
2.2NK cell Activity assay-Lactate Dehydrogenase (LDH) assay was used.
The spleen was removed aseptically and made into single cell suspensions as effector cells. Taking 100 mu L of target cells and effector cells respectively (the effective-target ratio is 50:1), and adding the target cells and the effector cells into a U-shaped 96-hole culture plate; target cells and culture solution are added into a natural release hole of the target cells by 100 mu L respectively, and target cells and 2.5 percent Triton are added into a maximum release hole of the target cells by 100 mu L respectively; all the above-mentioned materials are equipped with three composite holes, at 37 deg.C and 5% CO2Culturing for 4h in incubator, centrifuging 96-well culture plate at 1500r/min for 5min, sucking 100 μ L supernatant per well, placing in flat-bottomed 96-well culture plate, adding 100 μ L LDH matrix solution, reacting for 3min, adding 1mol/L HCl 30 μ L per well, and measuring optical density at 490nm of microplate readerValue (OD).
Figure BDA0002134407070000101
The NK cell activity of the test sample group is obviously higher than that of the control group, and the positive result of the experiment can be judged. The specific data are shown in Table 6.
TABLE 6 NK cell Activity assay results (
Figure BDA0002134407070000103
n＝10)
Figure BDA0002134407070000102
Figure BDA0002134407070000111
Note: p <0.05, indicating significant difference compared to control group; p <0.01, indicating a very significant difference compared to the control group.
3. And (3) analyzing an experimental result: table 6 shows test data on the influence of NK cell activity of mice, and it can be seen from experimental data that, compared with a control group, the medium and high dose groups of examples 6 to 9 of the present invention have significant differences compared with the control group, which indicates that the composition prepared in examples 6 to 9 of the present invention can improve NK cell activity of mice and has the effect of enhancing immunity; meanwhile, the low and medium dose groups in example 8 have significant difference compared with the control group, and the high dose group has significant difference compared with the control group, which indicates that the composition prepared in example 8 has higher NK cell activity and thus plays a better immune role.
Experimental example four, Effect of composition on mouse tail suspension immobility time
1. Experimental animals: KM mice, each half male and female, with the weight of 18-22 g;
2. the experimental method comprises the following steps:
2.1 experimental grouping and dosing: the recommended amount of the composition for human body is 1.1 g/kg-bw per person per day, and the composition is taken after being mixed with warm water. Mice experimental dose settings were low, normal, and high at 3 dose levels, i.e., 5.5, 11, and 33g/kg · bw per day (corresponding to 5, 10, and 30 times the recommended amount for humans, respectively), while control groups were set, and experimental mice were randomly batched by body weight, 5 groups per batch, and 10 mice per group.
The stomach is irrigated once a day with 0.2ml/10g · bw each time according to the experimental dose, normal control group is given with physiological saline with the same volume, and positive control group (fluoxetine hydrochloride 80mg/kg) is continuously given for 7 days.
2.2 Experimental methods:
after the last administration for 1h, the tail part of the mouse is attached to a horizontal stick at a position of 2cm, the head part of the mouse is about 5cm away from the table board, the two sides of the suspension are separated from the sight line of the animal by using partition plates, and the time that the tail of the mouse is suspended still within 6min is observed and recorded. The data are shown in Table 7.
TABLE 7 Effect of compositions on forced tail suspension immobility time in mice: (
Figure BDA0002134407070000112
n＝10)
Figure BDA0002134407070000121
Note: p <0.05, indicating significant difference compared to normal control group; p <0.01, indicating a very significant difference compared to the normal control group.
2.3 Experimental results: the composition can shorten forced tail suspension immobility time of mice, wherein the tail suspension immobility time of the low-dose groups of examples 6-9 is shortened, but has no significant difference compared with a normal control group; the middle dose groups of examples 6, 7 and 9 had significant differences compared to the normal control group, and the high dose groups had very significant differences compared to the normal control group; the middle and high dose groups of example 8 had very significant differences compared to the normal control group. The experimental results show that the composition of the invention can resist the depression symptom caused by forced tail suspension of mice.
The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.

Claims (8)

1. The composition with the brain strengthening/brain nerve conditioning effect is characterized by comprising the following components in parts by weight:
0.0002 to 0.0005 portion of folic acid;
2-4.5 parts of ginkgo leaves;
0.5 to 1.5 portions of galangal;
1-3 parts of white peony root;
0.5-1.5 parts of ligusticum wallichii;
3-5 parts of salvia miltiorrhiza;
1-3 parts of rhizoma anemarrhenae;
1-3 parts of wolfberry fruit;
the mass ratio of the ginkgo leaves to the galangal to the rhizoma anemarrhenae is 3:1: 2.
2. The composition with the brain strengthening/brain nerve conditioning effect is characterized by comprising the following components in parts by weight:
0.0002 to 0.0005 portion of folic acid;
2-4.5 parts of ginkgo leaves;
0.5 to 1.5 portions of galangal;
1-3 parts of white peony root;
0.5-1.5 parts of ligusticum wallichii;
3-5 parts of salvia miltiorrhiza;
1-3 parts of rhizoma anemarrhenae;
1-3 parts of wolfberry fruit;
3.5-5 parts of kudzu root;
2-4.5 parts of cassia seed;
2-4.5 parts of spina date seeds;
0.5-1.5 parts of fructus alpiniae oxyphyllae;
1-3 parts of gardenia;
1.7-2.5 parts of perilla leaf;
0.5-1.5 parts of platycladi seed;
4-7.5 parts of gynostemma pentaphylla;
0.5-1.5 parts of ginseng;
1-3 parts of semen cuscutae;
0.7-2 parts of schisandra chinensis;
0.5-1.5 parts of polygala root;
the mass ratio of the ginkgo leaves to the galangal to the rhizoma anemarrhenae is 3:1: 2.
3. A method of preparing a composition according to any one of claims 1 to 2, comprising the steps of:
(1) mixing the composition of any one of claims 1-2 except folic acid, extracting with water for three times, mixing the three filtrates, concentrating under reduced pressure, and drying to obtain Chinese medicinal extract;
(2) and (2) crushing and sieving the traditional Chinese medicine extract obtained in the step (1), and mixing the crushed extract with folic acid in a formula amount to obtain the composition with the effects of strengthening brain and conditioning cranial nerves.
4. The production method according to claim 3, wherein the concentration under reduced pressure in the step (1) is a concentration to a relative density of 1.05 to 1.25.
5. Use of a composition according to any one of claims 1-2 for the manufacture of a medicament for the treatment of brain tonic/cranial nerve modulation.
6. An oral preparation having brain strengthening/cranial nerve regulating effects, comprising the composition of any one of claims 1 to 2 and a pharmaceutically acceptable excipient.
7. The oral preparation according to claim 6, wherein the oral preparation is any one of a tablet, a capsule, a granule, a dripping pill and an oral liquid.
8. The oral preparation according to claim 6, wherein the excipient comprises any one of a disintegrant, a lubricant and a binder.
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