Coreopsis tinctoria extract for improving learning and memory disorders as well as preparation method and application thereof

Abstract

The invention relates to a senecio cineraria extract for improving learning and memory impairment and a preparation method and application thereof, and relates to the field of medicines. The coreopsis tinctoria extract is obtained by alcohol extraction of coreopsis tinctoria and solvent extraction. The solvent comprises at least one of ethyl acetate and n-butyl alcohol. The coreopsis tinctoria extract has a protection effect on neurons and can improve oxidative damage and apoptosis of neuronal cells, so that learning and memory disorders are improved and / or treated.

Description

Technical Field

[0001] The invention relates to the field of medicines, and in particular to a snow chrysanthemum extract for improving learning and memory disorders, and a preparation method and application thereof. Background Art

[0002] Learning and memory are one of the important functions of the human brain and a kind of high-level victorious activity in which the central nervous system participates. They are of great significance to people's learning and life. With the continuous increase of age, learning and memory will decline to varying degrees. Some of them are pathological, such as Alzheimer's disease, and some are physiological, such as physiological changes caused by aging. Nowadays, population aging is a common problem faced by the world, and more and more middle-aged and elderly people suffer from the decline of learning and memory. Therefore, how to prevent and improve learning and memory disorders related to aging at an early stage is one of the current research hotspots.

[0003] Snow chrysanthemum is native to western North America and is widely cultivated around the world. It contains a variety of active ingredients and has multiple effects such as lipid-lowering, blood pressure-lowering, anti-inflammatory, and anti-oxidation. It has been used as a tea for hundreds of years. However, as a natural medicine, snow chrysanthemum is a complex mixed system containing multiple ingredients. Its efficacy is the result of the synergistic effect of multiple active ingredients. In order to achieve a certain therapeutic effect, there are often problems such as large dosage and slow effect, which is not conducive to the development and application of modern products. Therefore, it is necessary to further extract, separate and screen the activity of natural medicines to obtain one or several groups of active components with fewer ingredients and higher activity for subsequent development and utilization. Summary of the invention

[0004] In view of the above problems, the present invention provides an application of a snow chrysanthemum extract in the preparation of a product for improving and / or treating learning and memory disorders. The snow chrysanthemum extract has a protective effect on neurons, can improve oxidative damage and apoptosis of neuronal cells, thereby improving and / or treating learning and memory disorders.

[0005] In order to achieve the above-mentioned purpose, the present invention provides an application of a snow chrysanthemum extract in the preparation of a product for improving and / or treating learning and memory disorders. The snow chrysanthemum extract is obtained by alcohol extraction of snow chrysanthemum and then extraction with a solvent; the solvent includes at least one of ethyl acetate and n-butanol.

[0006] The inventors found in the research process that although most of the active substances in the snow chrysanthemum can be extracted after the natural medicine, snow chrysanthemum, is extracted by ethanol and separated, the ethanol extract contains various compounds with complex components and unclear functions, which hinders its application. Therefore, the inventors extracted the snow chrysanthemum extract by using a solvent (specifically ethyl acetate and / or n-butanol) on the basis of ethanol extraction, and the obtained snow chrysanthemum extract was confirmed by experiments to have a protective effect on neurons, improve oxidative damage and apoptosis of neuronal cells, and thus improve and / or treat learning and memory disorders.

[0007] In one embodiment, the product comprises a medicine or a food.

[0008] In one embodiment, the food comprises health food.

[0009] In one embodiment, when the solvent includes ethyl acetate, the amount of snow chrysanthemum extract in the product is ≥ 200 mg·kg -1 ·d -1 ;

[0010] When the solvent includes n-butanol, the amount of snow chrysanthemum extract in the product is ≥ 250 mg·kg -1 ·d -1 .

[0011] The present invention also provides a method for preparing the snow chrysanthemum extract in the application, comprising the following steps: extracting the snow chrysanthemum with alcohol to obtain a snow chrysanthemum alcohol extract, and extracting the snow chrysanthemum alcohol extract with a solvent to obtain the snow chrysanthemum extract;

[0012] The solvent includes at least one of ethyl acetate and n-butanol.

[0013] In one embodiment, the alcohol extraction is achieved by a heat reflux method, and the reaction conditions of the heat reflux method include: using ethanol with a volume concentration of 60-80% as the extraction agent, the extraction temperature is 70-80°C, and the solid-liquid ratio is 1:(8-12).

[0014] In one embodiment, the alcohol extraction time is 3.5-4.5h.

[0015] In one embodiment, the extraction step comprises: dissolving the snow chrysanthemum alcohol extract in a solvent to obtain a snow chrysanthemum alcohol extract solution, extracting with a solvent, removing the solvent, and obtaining the snow chrysanthemum extract.

[0016] In one embodiment, the solvent comprises water.

[0017] In one embodiment, the amount of the snow chrysanthemum alcohol extract and the solvent is 1g: (700-800)mL.

[0018] The present invention also provides the snow chrysanthemum extract obtained by the preparation method and used for improving learning and memory disorders.

[0019] The present invention also provides a medicine for improving and / or treating learning and memory disorders, comprising a medicine active ingredient and a pharmaceutically acceptable excipient, wherein the medicine active ingredient is the snow chrysanthemum extract.

[0020] In one embodiment, the drug is in the form of oral solution, pills, granules, capsules, tablets, pellets, injections or powders.

[0021] The present invention also provides a food for improving and / or treating learning and memory disorders, comprising an active ingredient and a food additive, wherein the active ingredient is the snow chrysanthemum extract.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention discloses a snow chrysanthemum extract for improving learning and memory disorders, and a preparation method and application thereof. The snow chrysanthemum extract has a protective effect on neurons, can improve oxidative damage and apoptosis of neuronal cells, and thus improve and / or treat learning and memory disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a result diagram of the model induction condition experiment in the embodiment;

[0025] Figure 2 This is a result diagram of in vitro activity screening in the examples;

[0026] Figure 3 UPLC-MS positive ion chromatogram of EACC component analysis in the embodiment;

[0027] Figure 4 UPLC-MS negative ion chromatogram of EACC component analysis in the embodiment;

[0028] Figure 5 This is the UPLC-MS positive ion chromatogram of NBAC component analysis in the embodiment;

[0029] Figure 6 This is the UPLC-MS negative ion chromatogram of NBAC component analysis in the embodiment;

[0030] Figure 7 The figure is a measurement result diagram of the autonomous activity experiment in the embodiment, wherein: Figure 7 A is the result of activity frequency measurement, Figure 7 B is the result of the standing times measurement;

[0031] Figure 8 This is a measurement result diagram of the dark avoidance experiment in the embodiment, wherein: Figure 8A is the result of the incubation period measurement. Figure 8 B is the result of the error number determination;

[0032] Fig. 9 The figure is a measurement result diagram of the Morris water maze positioning cruise experiment in the embodiment;

[0033] Fig.10 The Morris water maze spatial exploration mouse swimming trajectory diagram in the embodiment, wherein: Fig.10 A is the swimming trajectory of mice in the blank group. Fig.10 B is the swimming trajectory of mice in the model group. Fig.10 C is the swimming trajectory of mice in the positive control group. Fig.10 D is the swimming trajectory of mice in the EACC high-dose group. Fig.10 E is the swimming trajectory of mice in the EACC medium-dose group. Fig.10 F is the swimming trajectory of mice in the EACC low-dose group. Fig.10 G is the swimming trajectory of mice in the NBAC high-dose group. Fig.10 H is the swimming trajectory of mice in the NBAC medium-dose group, Fig.10 I is the swimming trajectory of mice in the NBAC low-dose group;

[0034] Fig.11 The result diagram of Morris water maze space exploration experiment in the embodiment;

[0035] Fig.12 Graph showing the results of measuring oxidative stress indicators in mouse serum in the embodiment;

[0036] Fig.13 Graph showing the results of measuring oxidative stress indicators in mouse hippocampal tissue in the embodiment;

[0037] Fig.14 The figure is the staining result of the pathological section of the mouse hippocampus CA1 region in the embodiment, wherein: Fig.14 A is the blank group, Fig.14 B is the model group, Fig.14 C is the EACC high-dose group, Fig.14 D is the NBAC high-dose group;

[0038] Fig.15 The figure is the staining result of the pathological section of the mouse hippocampus CA3 region in the embodiment, wherein: Fig.15 A is the blank group, Fig.15 B is the model group, Fig.15 C is the EACC high-dose group, Fig.15 D is the NBAC high-dose group;

[0039] Fig.16 The Hochest apoptosis staining diagram (left) and apoptosis rate analysis result diagram (right) of the mouse hippocampus CA1 region in the embodiment are shown, wherein: Fig.16 A is the blank group, Fig.16 B is the model group, Fig.16 C is the EACC high-dose group, Fig.16 D is the NBAC high-dose group;

[0040] Fig.17 The Hochest apoptosis staining diagram (left) and apoptosis rate analysis result diagram (right) of the mouse hippocampus CA3 region in the embodiment are shown, wherein: Fig.17 A is the blank group, Fig.17 B is the model group, Fig.17 C is the EACC high-dose group, Fig.17 D is the NBAC high-dose group;

[0041] Fig.18 Graph showing the results of apoptosis-related protein expression determination in mouse hippocampal tissue in the examples. DETAILED DESCRIPTION

[0042] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0044] source:

[0045] Snow chrysanthemum (origin: Heilongjiang Province, China), piracetam (CAS#: 7491-74-9, Northeast Pharmaceutical Group Co., Ltd., >98%, Shenyang, China); D-galactose (CAS#: 59-23-4, Beijing Dingguo Changsheng Biotechnology Co., Ltd., >98%, Beijing, China); superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China); glutamate (Glu) and γ-aminobutyric acid (GABA) kits (Shanghai Longteng Bioscience Co., Ltd., Shanghai, China); PrimeScript with gDNA remover TM RT kit (TAKARA BIOINC., Japan); Premix Ex Taq TMII (TAKARA BIO INC., Japan).

[0046] Nexera ultra-high performance liquid chromatograph LC-30A (Shimadzu Corporation, Japan); TripleTOF5600+ mass spectrometer (AB SCIEX, USA); Morris water maze system (Chengdu Taiman Company, China); NanoDrop 2000 (Thermo Fisher Scientific, USA); ABI 2720 thermal cycler (Thermo Fisher Scientific, USA); ABI 7900 real-time PCR system (Thermo Fisher Scientific, USA).

[0047] Unless otherwise specified, the reagents, materials, and equipment used in this example are all commercially available; the experimental methods are all conventional experimental methods in the art unless otherwise specified.

[0048] Example

[0049] 1. Preparation of snow chrysanthemum extract.

[0050] The petroleum ether fraction (PEC), ethyl acetate fraction (EACC) and n-butanol fraction (NBAC) of snow chrysanthemum were extracted in descending order of polarity by using ethanol (70%) followed by solvent extraction.

[0051] The preparation method is as follows:

[0052] Take an appropriate amount of snow chrysanthemum, use 70% ethanol by volume as solvent, solid-liquid ratio 1:10, heat and reflux at 75°C for 2 times, each time for 2 hours, combine the filtrate, evaporate and weigh after concentration, and obtain the active extract of snow chrysanthemum. The active extract of snow chrysanthemum is extracted in batches, about 20g each time, 500mL of water is added, ultrasonic 20min dissolution, and then extracted with petroleum ether, ethyl acetate, and n-butanol in order from low to high polarity. Each solvent is extracted 3 times, 500mL each time, and after each concentrated solvent is combined, evaporated and weighed, the petroleum ether part (PEC), ethyl acetate part (EACC) and n-butanol part (NBAC) of snow chrysanthemum are obtained respectively.

[0053] 2. Activity screening experiment.

[0054] 1. D-galactose-induced PC12 cell injury model.

[0055] Take cells in the logarithmic growth phase and make them into a uniform cell suspension by repeated gentle pipetting. Then adjust the concentration to 4×10 4 pc·mL -1, inoculated in a 96-well plate, 100 μL of cell suspension was added to each well, and incubated in a CO2 incubator overnight; after 24 h, the culture medium of each well was aspirated, and 100 μL of the corresponding concentration of drug (prepared with DMEM culture medium containing 10% FBS) was added to each well of the drug-treated group, and the same volume of DMEM culture medium containing 10% FBS was added to the model group (MOD), and incubated in a CO2 incubator overnight; after 24 h, 100 μL of supernatant was aspirated from the model group and each drug-treated group, and 30 mg·mL -1 100 μL of D-galactose solution (prepared with DMEM culture medium containing 10% FBS) was added and the culture was continued for 24 h.

[0056] The preliminary modeling condition screening experiment proved that when the concentration was 30 mg·mL -1 When D-galactose was used, the damage degree of PC12 cells induced by D-galactose was moderate, and the average cell viability was 54.78%, which was suitable for drug administration research. -1 The concentration of D-galactose was used as the optimal induction condition for the PC12 cell model. The experimental results of the model induction conditions are shown in Figure 1 .

[0057] 2. Determine the effects of petroleum ether fraction (PEC), ethyl acetate fraction (EACC) and n-butanol fraction (NBAC) on neuronal cell viability.

[0058] Methods: According to the MTT method in the literature (Yan Yuhui, Li Shaoheng, Kong Liang, et al. Study on the neuroprotective effect of Angelica dahurica coumarin on Aβ-induced neuronal damage [J]. Liaoning Journal of Traditional Chinese Medicine, 2016, 43(08): 1714-1717.), the effects of petroleum ether fraction (PEC), ethyl acetate fraction (EACC) and n-butanol fraction (NBAC) on neuronal cell viability were determined, and the active fraction of snow chrysanthemum with neuroprotective effect was screened.

[0059] 3. UPLC-MS analysis.

[0060] Method: Take EACC and NBAC samples, weigh them accurately, add appropriate amount of ethyl acetate and n-butanol respectively, dissolve them by ultrasonic for 20 min, and then adjust the volume to 20 mg·mL -1 , centrifuged at 12000r / min for 10min as the test sample; 10μL of the supernatant of the two test samples were respectively injected, and the active parts of snow chrysanthemum were analyzed by UPLC-MS. The component composition information of the active parts of snow chrysanthemum was obtained after comprehensive analysis based on the search and comparison of the mass spectrum library and the matching degree and other information.

[0061] 3. Animal behavior experiments.

[0062] 1. Experimental animals.

[0063] 180 SPF-grade ICR mice (male; 4-6 weeks old, weighing 18-22 g) were purchased from Changchun Yitong Laboratory Animal Technology Co., Ltd. (license number: scxk-2018-0007). The mice were individually housed in cages at 20-24°C and 40-60% humidity, with free access to water and food under a 12-h day / night cycle. The experiment began after 3 days of laboratory adaptation feeding.

[0064] 2. Animal grouping and modeling.

[0065] One hundred and eighty male ICR mice were randomly divided into three groups: blank group (CON), model group (MOD), positive control group (piracetam, 300 mg kg -1 ·d -1 ), low, medium, and high doses of EACC groups (EACC-L: 100 mg kg -1 ·d -1 , EACC-M: 200 mg kg -1 ·d -1 、EACC-H:400mg·kg -1 ·d -1 ), low, medium, and high dose NBAC groups (NBAC-L: 125 mg kg -1 ·d -1 、NBAC-M:250mg·kg -1 ·d -1 、NBAC-H:500mg·kg -1 ·d -1 Except for the blank group, mice in other groups were subcutaneously injected with 300 μg / g -1 D-galactose, blank group mice were subcutaneously injected with an equal volume of saline every day. At the same time, mice in all groups were gavaged with different drugs once a day, and the CON group and MOD group were given an equal volume of distilled water. The drugs were administered continuously for 96 days after the start of the experiment.

[0066] 3. Investigate the improving effect of the active parts of snow chrysanthemum on learning and memory disorders in mice.

[0067] Methods: From the 85th to the 95th day, the spontaneous activity test, dark avoidance test and Morris water maze test were carried out to investigate the improving effect of the active fraction of snow chrysanthemum on the learning and memory disorders of mice.

[0068] On the 85th day, 30 minutes after administration, the mouse autonomous activity experiment was conducted to examine the differences in the autonomous activity behaviors of the animals in each group. Each time, 6 mice were placed in the autonomous activity tester at the same time, 1 in each compartment, and the recording time was set to 5 minutes after the top cover was closed. The instrument will automatically record the number of activities (horizontal activities) and the number of standing times (upright movements) of the mice within 5 minutes.

[0069] 30 minutes after drug administration on the 87th and 88th days, a dark avoidance experiment was conducted. The 87th day of drug administration was the training stage. Six mice were placed in the mouse dark avoidance instrument at the same time to adapt for 3 minutes. At this time, the gate between the light and dark chambers was opened, and no current was passed. The mice moved freely between the light and dark chambers. Then the mice were placed in the light chamber, the gate was closed, and a current of 36V and 0.5mA was passed to the dark chamber. The gate was opened, and the instrument automatically started timing. The time when the mouse first crossed the gate and entered the dark chamber was recorded as the latency. At the same time, the number of errors of the mouse entering the dark chamber within 5 minutes was recorded. After 5 minutes, the mouse was taken out and the training ended. The 88th day of drug administration was the formal experimental stage. The experimental method was the same as the power-on training. The latency (s) and number of errors of each group of mice were recorded, and compared and analyzed with the test results on the 87th day.

[0070] On the 90th to 95th day of drug administration, 30 minutes after drug administration, Morris water maze test was performed according to the method in the literature (①Morris R. Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods. 1984, 11(1): 47-60. ②Vorhees CV, Williams MT. Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc. 2006, 1(2): 848-858.).

[0071] 4. Experiment on oxidative damage and apoptosis of mouse hippocampal neurons.

[0072] 1. Using tissue samples obtained from animal experiments, the morphology of neuronal cells in the CA1 and CA3 regions of the mouse hippocampus was observed by HE section staining.

[0073] 2. According to the instructions of the kit, the WST-1 method, colorimetric method and TBA method were used to measure the activity of SOD and GSH-Px and the content of MDA in serum and hippocampal tissue, so as to investigate the protective effect of EACC and NBAC on oxidative damage of mouse hippocampal neurons.

[0074] 3. Take the brain tissue slices of mice and wash them with PBS for 3 times, 5 minutes each time. According to the method in the kit instructions, the brain tissue slices were stained with Hochest33258, and the apoptosis of neuronal cells in the CA1 and CA3 regions of the mouse brain tissue hippocampus was observed under a fluorescence microscope after sealing.

[0075] Five sections of corresponding positions were selected for staining for each mouse. Five fields of view were selected in the CA1 and CA3 regions of the hippocampus of each section. The total number of neurons and the number of apoptotic neurons in each field of view were manually counted, and the apoptotic rate of neurons was calculated. The calculation formula is as follows:

[0076] Cell apoptosis rate (%) = number of apoptotic neurons / total number of neurons × 100%

[0077] The mean apoptosis rate of neurons in the CA1 and CA3 regions of each slice was calculated (5 fields of view), and the mean apoptosis rate of neurons in the CA1 and CA3 regions of the hippocampus of each mouse was calculated (5 slices); statistical comparison was performed among the experimental groups.

[0078] To investigate the inhibitory effects of EACC and NBAC on neuronal apoptosis in CA1 and CA3 regions of the mouse hippocampus.

[0079] 4. Take the whole brain of some mice in each group, separate the hippocampal tissue on ice, cut it into pieces, add protease inhibitors and RIPA lysis buffer, grind it thoroughly with a homogenizer, centrifuge it at 4℃, 13000r / min for 10min, and take the supernatant. Use Western Blot to analyze the expression of apoptosis-related proteins Bcl-2, Bax and Cleaved Casepase-3 in hippocampal tissue.

[0080] 5. Statistical analysis.

[0081] The data of this example are expressed as mean ± standard deviation, and one-way analysis of variance was performed using SPSS 19.0 statistical software, and the results between groups were compared using t-test. P < 0.05 or P < 0.01 was considered statistically significant.

[0082] 6. Experimental results.

[0083] 1. Results of activity screening experiments.

[0084] In EACC and NBAC, the PC12 cell viability in the high-dose group was significantly higher than that in the MOD group (P < 0.01); while the cell viability in each dose group of PEC was not significantly different from that in the MOD group (P > 0.05). It can be seen that EACC and NBAC have a certain protective effect on neuronal cells in vitro and are the active parts of snow chrysanthemum. After UPLC-MS determination results were retrieved by mass spectrometry database and comprehensively analyzed and screened, it was shown that EACC and NBAC contained 39 and 26 main active ingredients, respectively.

[0085] The results of in vitro activity screening are shown in Figure 2 UPLC-MS chromatograms are shown in Figure 3-Figure 6 .

[0086] 2. Results of animal behavior experiments.

[0087] In the spontaneous activity test, there was no significant difference in the number of activities and standing times of mice in each group (P>0.05). In the dark avoidance test, high doses of EACC and medium and high doses of NBAC can significantly prolong the latency of model mice and reduce the number of errors (P<0.05), and medium doses of EACC also have a certain improvement effect on the latency and number of errors of model mice (P>0.05). In the positioning cruising test of Morris water maze, medium and high doses of EACC and NBAC can significantly shorten the latency of model mice to find the escape platform; at the same time, in the spatial exploration test, medium and high doses of EACC and NBAC can significantly increase the percentage of model mice entering the target quadrant, the time they stay in the target quadrant, and the number of times they enter the effective area and the original platform area.

[0088] The results of the spontaneous activity experiment are shown in Figure 7 ; The results of the dark avoidance experiment are shown in Figure 8 Morris water maze test results are shown in Figure 9-11 .

[0089] 3. Experimental results of oxidative damage and apoptosis of mouse hippocampal neurons.

[0090] EACC and NBAC can effectively improve the oxidative damage of neurons in the CA1 and CA3 regions of the hippocampus of mice induced by D-galactose by upregulating the activity levels of SOD and GSH-Px in the serum and hippocampal tissue of model mice and reducing the content of MDA (P<0.01, P<0.05), maintaining the basic morphology of cells. At the same time, EACC and NBAC have a significant inhibitory effect on the apoptosis of neurons in the CA1 and CA3 regions of the hippocampus of mice induced by oxidative damage, and the apoptosis rate is significantly lower than that of the MOD group (P<0.01). In addition, compared with the MOD group, the expression of Bcl-2 in the hippocampal tissue of mice in the EACC and NBAC administration groups was significantly increased (P<0.01), the expression of Bax and Cleaved Casepase-3 was significantly decreased (P<0.01, P<0.05), and the ratio of Bax / Bcl-2 was downregulated (P<0.01), close to the level of the CON group.

[0091] The results of oxidative stress related indicators are shown in Figure 12-13 ; The staining results of pathological sections of mouse hippocampal CA1 and CA3 regions are shown in Figure 14-15 ; The results of apoptosis staining in CA1 and CA3 regions of mouse hippocampus are shown in Figure 16-Figure 17 ; The results of apoptosis-related protein expression in mouse hippocampus tissue are shown in Fig.18

[0092] 8. Summary

[0093] 1. The ethyl acetate and n-butanol parts are the active parts of snow chrysanthemum that have neuron-protective effects, and have a significant improvement effect on D-galactose-induced learning and memory disorders in mice.

[0094] 2. The active part of snow chrysanthemum has a protective effect on oxidative damage and apoptosis of neuronal cells in the hippocampus of the brain, providing a physiological basis for the improvement of learning and memory disorders in mice.

[0095] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0096] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A use of a snow chrysanthemum extract in the preparation of a product for improving and / or treating learning and memory disorders, wherein the snow chrysanthemum extract is obtained by alcohol extraction of snow chrysanthemum and then extraction with a solvent; the solvent comprises at least one of ethyl acetate and n-butanol.

2. The use according to claim 1, characterized in that: The product includes a medicine or a food.

3. The use according to claim 1, characterized in that: When the solvent includes ethyl acetate, the amount of snow chrysanthemum extract in the product is ≥ 200 mg·kg -1 ·d -1 ; When the solvent includes n-butanol, the amount of snow chrysanthemum extract in the product is ≥ 250 mg·kg -1 ·d -1 .

4. A method for preparing the snow chrysanthemum extract for use in any one of claims 1 to 3, characterized in that: The following steps are involved: Extracting snow chrysanthemum with alcohol to obtain snow chrysanthemum alcohol extract, and extracting the snow chrysanthemum alcohol extract with a solvent to obtain a snow chrysanthemum extract; The solvent includes at least one of ethyl acetate and n-butanol.

5. The preparation method according to claim 4, characterized in that: The alcohol extraction is achieved by a heat reflux method, and the reaction conditions of the heat reflux method include: using ethanol with a volume concentration of 60-80% as an extractant, an extraction temperature of 70-80°C, and a solid-liquid ratio of 1:(8-12).

6. The preparation method according to claim 4, characterized in that: The extraction step comprises: dissolving the snow chrysanthemum alcohol extract in a solvent to obtain a snow chrysanthemum alcohol extract solution, extracting with a solvent, removing the solvent, and obtaining the snow chrysanthemum extract.

7. The snow chrysanthemum extract for improving learning and memory disorders obtained by the preparation method according to any one of claims 4 to 6.

8. A drug for improving and / or treating learning and memory disorders, characterized in that: The invention comprises a pharmaceutically active ingredient and a pharmaceutically acceptable excipient, wherein the pharmaceutically active ingredient is the snow chrysanthemum extract as claimed in claim 7.

9. The drug according to claim 8, characterized in that The dosage form of the medicine is oral liquid, pill, granule, capsule, tablet, dripping pill, injection or powder.

10. A food for improving and / or treating learning and memory disorders, characterized in that: The invention comprises an active ingredient and a food additive, wherein the active ingredient is the snow chrysanthemum extract according to claim 7.

Images