Dendrobium huoshanense extract and application thereof in preparation of medicine for preventing and / or treating depression

Abstract

The invention discloses a dendrobium huoshanense extract and application of the dendrobium huoshanense extract in preparation of a medicine for preventing and / or treating depression, and belongs to the technical field of biological medicine. The dendrobium huoshanense extract is obtained through reflux extraction by using an alcohol-water mixed solution, and the volume concentration of alcohol in the alcohol-water mixed solution is 50%-90%. The reflux extraction conditions are as follows: the temperature is 70-90 DEG C, and the time is 1-4 hours. According to behavioral data analysis of mouse sweet water preference, tail suspension test, forced swimming test, open field test, elevated cross labyrinth test and the like, the dendrobium huoshanense extract has a remarkable anti-depression effect, has a good effect of preventing and / or treating depression, and has a good application prospect in prevention and / or treatment of depression.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a Huoshan dendrobium officinale extract and application thereof in preparing a medicine for preventing and / or treating depression. Background Art

[0002] Major depressive disorder (MDD) is a serious mental illness characterized by low mood, lack of motivation, despair and inability to feel happiness, slow thinking and movement, accompanied by sleep disorders, loss of appetite, etc. MDD is one of the main causes of disability worldwide. Although many advances have been made in the drug treatment of depression, more than 50% of patients do not improve after first-line antidepressant treatment. Mechanism studies have shown that the pathology of depression is closely related to genetics, neurotransmitter systems, brain-derived neurotrophic factor, hypothalamus-pituitary-adrenal (HPA) axis and inflammatory factors, such as most of the antidepressants currently used in clinical practice (fluoxetine, paroxetine, fluvoxamine, sertraline and citalopram, etc.). In general, these drugs have limitations such as slow onset, low antidepressant efficacy, large toxic side effects, and high prices. It is precisely because of the defects of existing antidepressant drugs and the urgency of market demand that domestic and foreign scholars have turned their attention to Chinese herbal medicine. Natural medicines with antidepressant effects are mostly found in the family of Garcinia, Rubiaceae, Ginkgo, Annonaceae, Aristolochiaceae, Dicentraceae, Cucurbitaceae, Palmaceae, Leguminosae, Pinaceae, Asteraceae, etc. Relatively more studied plants include: Hypericum perforatum (St. John's wort, Hypericum perforatum), Morinda officinalis, Acanthopanax senticosus, Ginkgo, Valerian, Areca catechu, Astragalus membranaceus, etc. In recent years, antidepressant drugs derived from Chinese herbal medicines have been launched one after another, such as the Thirteen Flavors Depression-Relief Capsules (mainly containing Tripterygium wilfordii, Curcuma zedoaria, Corydalis yanhusuo, etc.) produced by Guangdong Zhongsheng, Shugan Jieyu Capsules (Hypericum perforatum, Acanthopanax senticosus) produced by Chengdu Kanghong, Zhenling Jieyu Capsules (mainly containing Paeonia lactiflora, Angelica sinensis, Bupleurum, etc.) produced by Shanxi Maidi, Morinda officinalis Oligosaccharide Capsules (Morinda officinalis) produced by Beijing Tongrentang, and other large antidepressant Chinese medicine compound prescriptions. Therefore, there is great potential in finding natural antidepressants from Chinese herbal medicine.

[0003] Huoshan Dendrobium (Dendrobium huoshanense), commonly known as "rice dendrobium", is a perennial herbaceous plant of the genus Dendrobium in the Orchidaceae family. It has a long history and was first recorded in detail in "Supplement to Compendium of Materia Medica". It is known as the "first of the nine immortal herbs". It is listed as a top-grade product in "Shennong's Herbal Classic": "It tastes sweet, flat, and non-toxic. It is mainly used to treat injuries, remove numbness, lower qi, nourish the five internal organs, and treat asthenia, thinness, and strengthen yin. Long-term use thickens the stomach and intestines"; "Famous Doctors' Records" describes Huoshan Dendrobium as "nourishing the five internal organs, thickening the stomach and intestines, strengthening muscles and bones, strengthening yin and benefiting essence, removing numbness and lowering qi, calming the mind and eliminating shock, and lightening the body and prolonging life". Among them, "calming the mind and eliminating shock" is associated with depression. Domestic and foreign researchers have studied the chemical composition and pharmacological activity of its stems. Its main chemical components are polysaccharides, flavonoids, phenanthrenes, biphenyls, alkaloids, etc. Its main pharmacological activities are anti-tumor, antioxidant, lipid regulation, hypoglycemic, anti-inflammatory, and strengthening the intestinal physiological barrier. However, there are no reports on its antidepressant activity. Summary of the invention

[0004] The purpose of the present invention is to provide a Huoshan Dendrobium officinale extract and its use in the preparation of a drug for preventing and / or treating depression, so as to solve the problems existing in the above-mentioned prior art. The present invention finds that the Huoshan Dendrobium officinale extract can effectively alleviate depression symptoms.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] Technical solution 1: A Huoshan Dendrobium extract, which is obtained by reflux extraction using an alcohol-water mixture, wherein the volume concentration of alcohol in the alcohol-water mixture is 50%-90%.

[0007] Furthermore, the conditions of the reflux extraction include: temperature of 70-90° C. and time of 1-4 h.

[0008] In the present invention, in order to further improve the antidepressant effect of the extract, the volume concentration of alcohol in the alcohol-water mixture is preferably 70%.

[0009] Furthermore, the preparation method of the Huoshan Dendrobium extract includes: firstly subjecting the stems of the Huoshan Dendrobium to reflux extraction with a mixture of alcohol and water, and then reducing the pressure and concentrating until there is no alcohol taste to obtain a suspended aqueous solution; extracting with petroleum ether, ethyl acetate, and n-butanol in sequence, and finally obtaining solutions of each component, and reducing the pressure, concentrating, and drying to obtain the Huoshan Dendrobium extract.

[0010] In the present invention, in order to further improve the antidepressant effect of the extract, the preparation method of the Huoshan Dendrobium extract comprises:

[0011] S1. Pretreatment: crushing the stems of Dendrobium huoshanense;

[0012] S2, extraction: using an alcohol-water mixture to reflux extract the stems of Dendrobium huoshanense, repeating this operation three times, combining the filtrates, decompressing and concentrating until there is no alcohol taste, and obtaining a water suspension of Dendrobium huoshanense;

[0013] S3, first extraction: add an equal volume of petroleum ether for extraction, repeat the operation three times, combine the upper liquid layer to obtain the petroleum ether layer;

[0014] S4, second extraction: take the lower layer liquid of step S3, add an equal volume of ethyl acetate, repeat the operation three times, combine the upper layer liquid, and obtain the ethyl acetate layer;

[0015] S5, third extraction: take the lower layer liquid of step S4, add an equal volume of n-butanol, repeat the operation three times, combine the upper layer liquid to obtain the n-butanol layer, and the lower layer is the water layer of the Huoshan Dendrobium extract;

[0016] S6, drying: optionally, removing the organic solvent and water in each component to obtain the Dendrobium huoshanense extract.

[0017] In the present invention, step S1 includes crushing the Dendrobium huoshanense. There is no special requirement for the crushing method, and conventional methods in the art can be used. There is no special requirement for the crushing time, and for example, it can be any range of 5-15 minutes.

[0018] In the present invention, step S1 also includes screening, and there is no special requirement for the mesh size of the screening, for example, it can be 10-60 meshes.

[0019] In the present invention, in order to further improve the antidepressant effect of the extract; in step S2, the amount of alcohol used accounts for 50-90% of the total amount of alcohol and water, 50-90% is the volume concentration, preferably 70%; in step S2, the conditions of the reflux extraction include: the temperature is 70-90°C, preferably 85°C; the time of the reflux extraction is 1-4h, preferably 2h.

[0020] In the present invention, steps S3, S4 and S5 further include extraction, and there is no special requirement for the extraction, and conventional methods in the art may be used.

[0021] In the above preferred embodiment of the present invention, in order to better remove the solvent, the drying method in step S6 is reduced pressure drying.

[0022] The present invention also provides the use of the Huoshan Dendrobium officinale extract in preparing a medicine for preventing and / or treating depression.

[0023] Further, the depression is caused by chronic unpredictability and mild stress.

[0024] Furthermore, the depression includes at least one of depression accompanied by anxiety and depression accompanied by mania.

[0025] In the present invention, the depression is not particularly limited, and can be at least one of mild depression, depression without psychotic symptoms, depression with psychotic symptoms, and recurrent depression. The depression can be at least one of depression with anxiety and depression with mania.

[0026] In the present invention, the extract can be prepared into various forms such as decoction, pills, granules, injections, ointments, tablets, capsules, syrups, drops, etc. when used to prepare drugs for preventing and / or treating depression. The drugs in the above various dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0027] In the present invention, when the extract is used to prepare a drug for preventing and / or treating depression, in order to solve the problems of formability, effectiveness, stability, safety, etc. of the preparation, one or more pharmaceutically acceptable carriers such as absorption enhancers, flavoring agents, adhesives, lubricants, antioxidants, diluents, disintegrants, surfactants, adsorption carriers, etc. may be added when necessary.

[0028] In the present invention, when the extract is used to prepare a drug for preventing and / or treating depression, the drug can be administered orally, sublingually, intravenously, intramuscularly, nasogastricly, or other common administration methods.

[0029] The present invention discloses the following technical effects:

[0030] The present invention has found that the petroleum ether layer, ethyl acetate layer, n-butanol layer and water layer of the Huoshan Dendrobium extract can improve the depressive-like behavior of mice subjected to chronic unpredictable mild stress, and at the same time regulate the neuroinflammatory response induced by chronic unpredictable mild stress and improve nerve damage. The present invention discloses that the Huoshan Dendrobium extract has a good antidepressant effect and has a good application prospect in the prevention and / or treatment of depression. The present invention has found that the Huoshan Dendrobium extract can effectively alleviate depressive symptoms, specifically: in a depressive-like mouse model induced by chronic unpredictable mild stress, it can effectively alleviate the decline in the preference of the depressed model mice for sucrose water, shorten the immobility time of the forced swimming and tail suspension of the depressed model mice, increase the amount of exercise in the central area of ​​the depressed model mice in the open field, improve the novel exploration behavior of the cross elevated maze of the depressed mice, improve the level of pro-inflammatory cytokines in the depressed mice, and alleviate neuronal damage. It can be seen that the present invention provides a new idea for the use of Huoshan Dendrobium for the prevention and / or treatment of depression. DETAILED DESCRIPTION

[0031] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0032] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0033] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0034] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0035] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0036] In the present invention, when preparing the Huoshan Dendrobium extract, the amount of alcohol used accounts for 50-90% of the total amount of alcohol and water, 50-90% is the volume concentration, and the following examples take 70% as an example; the conditions for reflux extraction include: the temperature is 70-90°C, and the following examples take 85°C as an example; the reflux extraction time is 1-4h, and the following examples take 2h as an example. When the Huoshan Dendrobium is crushed, the present invention has no special requirements for the crushing method, and the conventional method in the field can be used. The crushing time has no special requirements and can be in any range of 5-15min; the present invention has no special requirements for the sieve mesh, which can be 10-60 mesh; and the drying is reduced pressure drying.

[0037] Example 1

[0038] (1) Preparation method of Huoshan Dendrobium officinale extract

[0039] The dried stem of Dendrobium huoshanense is taken and crushed, and then 20 times the volume of ethanol solution with a volume concentration of 70% is added, and reflux extraction is performed at a temperature of 85°C, and each reflux is performed for 2 hours. This operation is repeated 3 times to obtain 3 extracts respectively, and the extracts are combined and concentrated under reduced pressure until the ethanol is completely volatilized (without alcohol taste) to obtain a water suspension of Dendrobium huoshanense; then, an equal volume of petroleum ether is used for extraction, and this operation is repeated 3 times, and the extracts of the upper petroleum ether are combined; the extracts are concentrated under reduced pressure and dried, and the obtained powder is the petroleum ether component of the extract of Dendrobium huoshanense, and is stored in a refrigerator at 4°C for standby use; then, the lower aqueous suspension solution is treated with an equal volume of acetic acid Ethyl ester extraction, extraction for 3 times, combined the upper ethyl acetate extract, the extract was concentrated under reduced pressure and dried, the obtained powder was the ethyl acetate component of Huoshan Dendrobium extract, and placed in a 4°C refrigerator for standby use; then the lower aqueous suspension solution was extracted with an equal volume of water-saturated n-butanol, extracted 3 times, combined the upper water-saturated n-butanol extract, the extract was concentrated under reduced pressure and dried, the obtained powder was the n-butanol component of Huoshan Dendrobium extract, and placed in a 4°C refrigerator for standby use; finally, the lower aqueous suspension solution was collected, concentrated under reduced pressure and dried, the obtained powder was the water layer component of Huoshan Dendrobium extract. The positive drug selected was fluoxetine, a commonly used antidepressant drug in clinical practice, which is fluoxetine hydrochloride produced by Shanghai McLean Biochemical Technology Co., Ltd.

[0040] (2) Animal experiments

[0041] The animals used in the present invention are male mice aged 6-8 weeks. The animals and feeds are purchased from the Experimental Animal Center of Henan Sikebes Biotechnology Co., Ltd., SPF-grade ICR mice, and the experimental animal quality certificate number is SCXK (Yu) 2020-0005; the animals are kept in an ordinary animal room with a temperature of 24±2°C, a humidity of 55%±5%, a light / dark cycle of 12 hours, standard feed, and free access to food and water. The protocol was approved by the Animal Ethics and Use Committee of Wanxi University.

[0042] (3) Experimental modeling and grouping

[0043] The classic chronic unpredictable mild stress method was used to model depression in mice. This method is similar to the pathogenesis of human depression, that is, depression is induced by adverse environment and other unfavorable factors. It is very suitable for mechanism research and drug screening. The types of chronic unpredictable mild stress include: (1) swimming in 30℃ warm water for 10 minutes; (2) swimming in 8℃ cold water for 5 minutes; (3) gently clamping the mouse tail for 1 minute; (4) day / night reversal for 24 hours; (5) cage tilted 45° for 24 hours; (6) noise interference for 5 minutes; (7) keeping the cage moist for 12 hours (water: 200mL / cage); (8) fasting for 12 hours; (9) fasting for 12 hours; each stimulus should be selected randomly, and the interval between the same stimulus should be greater than 7 days.

[0044] Grouping: After adaptive feeding, the mice were randomly divided into blank control group (Control, 0.5% CMCNa aqueous solution) by gavage; model control group (Model, 0.5% CMCNa aqueous solution) by gavage; positive drug fluoxetine group (FLU, 10.0 mg / kg dissolved in 0.5% CMCNa aqueous solution) by gavage; Huoshan Dendrobium huoshanense extract petroleum ether group (DHPE, 31.7 mg / kg (crude drug amount 4 g / kg) dissolved in 0.5% CMCNa aqueous solution) by gavage. a aqueous solution); ethyl acetate group of Dendrobium huoshanense extract (DHEA, 40 mg / kg (raw drug amount 4 g / kg) dissolved in 0.5% CMCNa aqueous solution); n-butanol group of Dendrobium huoshanense extract (DHnB, 128 mg / kg (raw drug amount 4 g / kg) dissolved in 0.5% CMCNa aqueous solution); water layer group of Dendrobium huoshanense extract (DHW, 565 mg / kg (raw drug amount 4 g / kg) dissolved in 0.5% CMCNa aqueous solution). The mouse depression model was established in combination with chronic unpredictable mild stress, and behavioral data such as sugar water preference, tail suspension, forced swimming, open field, and elevated maze were used as detection standards to determine the establishment of the depression model.

[0045] (4) Animal administration

[0046] After the model was established, the drug was administered by gavage for 21 consecutive days, and behavioral experiments were started after the last administration. Hippocampal tissue and blood were taken from mice for biochemical tests. In order to avoid interference from external factors, all behavioral experiments were conducted under conditions that ensured that the environment was not affected.

[0047] (5) Sugar water preference experiment

[0048] The principle of the sucrose preference test (SPT) is to prevent depressed mice from showing a preference for sucrose water like normal mice, reflecting the lack of pleasure. In order to change the drinking habits of mice, sucrose water training was performed on the third day of the adaptation period of mice. The specific method was: on the third day, two bottles of 1% sucrose water were given at the same time for 24 hours, and on the fourth day, one bottle of 1% sucrose water and one bottle of ordinary distilled water were given for 12 hours each. 12 hours before the formal sucrose preference experiment, fasting and water deprivation were required. Free drinking water was allowed for 12 hours, and then the consumption of sucrose water and distilled water was calculated. From the beginning of modeling, each group of mice underwent sucrose water preference experiments on days 0, 14, and 35, respectively.

[0049] Sucrose preference rate (%) = sucrose consumption / (sucrose consumption + tap water consumption) × 100%

[0050] (6) Tail suspension test

[0051] Tail suspension test (TST) is a classic method that can quickly evaluate the efficacy of antidepressants, stimulants, and sedatives. After the last administration of drugs to mice, a piece of string was fixed to the tail of the mouse 1 cm with tape, and the other end was fixed to a horizontal bar with a hook, and the mouse was fixed 2 cm from the floor. Each mouse was observed for 6 minutes, with the first 2 minutes of adaptation and the immobility time of the mouse recorded for the last 4 minutes. The mouse was considered immobile only when there was no escape and struggling behavior.

[0052] (7) Forced swimming test

[0053] Forced swimming test (FST) is mainly used in the study of antidepressant, sedative and analgesic drugs. This experimental method is a behavioral despair experimental method. Mice were placed in beakers filled with water (15 cm in diameter, 25 cm in height) at a water temperature of 24±2°C. Each mouse was forced to swim for 6 minutes, and the first 2 minutes were used to adapt to the environment. The immobility time of the mouse was recorded in the next 4 minutes. When the mouse stopped struggling and floated on the water, or when the limbs trembled slightly and the head floated above the water, it was considered to be in an immobile state.

[0054] (8) Open field test

[0055] The Open Field Test (OFT) is a behavioral test that is often used to evaluate the exploratory behavior, anxiety level, and motor ability of animals (especially mice and rats) in a new environment. In a quiet, dark environment, place the mouse in the center of the bottom of the box. Each mouse is free to explore for 2 minutes before the experiment. The activity of the mouse is then recorded by infrared video tracking software, and the following data are statistically calculated based on the experimental results: movement time, time to cross the central area. After each animal experiment, in order to eliminate the influence of animal odor on the activity of the next mouse, the square observation cage is cleaned with 70% ethanol solution and the feces is wiped dry. Excluding the free exploration time, the activity of each mouse is recorded for 4 minutes.

[0056] (9) Elevated plus maze test

[0057] The elevated plus maze test (EPMT) reflects the animal's anxiety by creating a conflicting state between the animal's exploration of a new environment and its fear of the suspended open arms. The elevated plus maze consists of a cross formed by the intersection of closed arms and open arms. The mouse is placed in the center of the cross. The mouse is allowed to explore freely for 2 minutes before the test. Infrared video tracking software is used to record the following data on the mouse's activities: the time the mouse stays in the open arm and the time the mouse moves. After each animal test, in order to eliminate the influence of animal odor on the next test, the equipment is cleaned with 70% ethanol solution and the feces is wiped dry. Excluding the free exploration time, each mouse is recorded for 4 minutes.

[0058] (10) ELISA method for determination of inflammatory factors

[0059] After the mice were anesthetized with chloral hydrate (3 mg / kg), the hippocampus was removed, 0.2 mL of phosphate buffered saline (PBS, pH = 7.4) was added to 20 mg of hippocampal tissue, and after complete homogenization, the supernatant was carefully collected into an Eppendorf tube; the mouse blood was centrifuged at 3000 rpm for 10 minutes, and the supernatant was carefully collected into an Eppendorf tube. The relevant experimental operations were carried out strictly in accordance with the instructions provided by the manufacturer of the ELISA kit.

[0060] The experimental data of each experiment are detailed in Tables 1 to 7.

[0061] Table 1 Content ratio of different components in Huoshan Dendrobium officinale extract (%)

[0062] Group Proportion (%) Potential ingredients Petroleum ether layer 0.79％ Volatile oils, resins, fats, etc. Ethyl acetate layer 1.0％ Free flavonoids, coumarins, etc. n-Butanol layer 3.2％ Some alkaloids, flavonoid glycosides, anthraquinone glycosides, triterpenes, etc. Water layer 14.12％ Water-soluble alkaloids, flavonoids, lignans, etc.

[0063] Table 2 Preference rate of sucrose water consumption in mice subjected to chronic unpredictable mild stress (%), n=12

[0064]

[0065] Note: Compared with the normal group, #p<0.05, ##p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0066] The results show that after the treatment with different components of the Huoshan Dendrobium extract obtained by the present invention, the animals' preference for sucrose water is restored, among which the water layer component of the Huoshan Dendrobium extract has the best effect, suggesting that the Huoshan Dendrobium extract has an improving effect on depression.

[0067] Table 3 TST immobility time (s) of mice subjected to chronic unpredictable mild stress, n=12

[0068] Group Immobility time (x±s,s) Control 55.4±6.22 Model <![CDATA[105.6±8.16 ## ]]> FLU <![CDATA[65.8±3.48 ** ]]> DHPE <![CDATA[80.0±5.54 ** ]]> DHEA <![CDATA[78.3±4.38 ** ]]> HkDJ <![CDATA[88.6±6.60 * ]]> DHW <![CDATA[64.2±2.72 ** ]]>

[0069] Note: Compared with the normal group, #p<0.05, ##p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0070] The results showed that compared with the animals in the normal group, the animals in the model group had a significantly suppressed desire for physical activity for survival, as shown by a significantly prolonged tail suspension immobility time. After treatment with different components of the Huoshan Dendrobium extract, the animals' desire for physical activity for survival increased, and the tail suspension immobility time was significantly shortened, among which the water layer component of the Huoshan Dendrobium extract had the best effect. This suggests that the Huoshan Dendrobium extract has an improving effect on depression.

[0071] Table 4 FST immobility time (s) of mice subjected to chronic unpredictable mild stress, n=12

[0072] Group Immobility time (x±s,s) Control 123.5±12.53 Model <![CDATA[163.8±16.08 ## ]]> FLU <![CDATA[130.5±15.33 ** ]]> DHPE <![CDATA[148.0±14.48 ** ]]> DHEA <![CDATA[147.3±14.66 ** ]]> HkDJ <![CDATA[137.8±13.65 * ]]> DHW <![CDATA[131.6±13.72 ** ]]>

[0073] Note: Compared with the normal group, #p<0.05, ##p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0074] The results showed that compared with the normal group of animals, the model group animals had a significantly suppressed desire to swim for survival under water stress, which was manifested by a significantly prolonged immobility time. After treatment with different components of Huoshan Dendrobium extract, the swimming activity of mice increased, which was manifested by a significantly shortened immobility time, among which the water layer component of Huoshan Dendrobium extract had the best effect. This suggests that Huoshan Dendrobium extract has an improving effect on depression.

[0075] Table 5 OFT activity status of mice subjected to chronic unpredictable mild stress, n=12

[0076]

[0077] Note: Compared with the normal group, #p<0.05, ##p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0078] The results showed that compared with the normal group animals, the total movement distance and central area distance of the model group animals in the open field were significantly shortened, and the central area residence time was reduced. After treatment with different components of Huoshan Dendrobium extract, the total movement distance and central area distance of the mice in the open field increased significantly, and the central area residence time was significantly prolonged, among which the water layer component of Huoshan Dendrobium extract had the best effect. This shows that Huoshan Dendrobium extract can increase the amount of exercise and central area exploration ability of depression model mice in the open field, and improve the overall activity state of depression mice.

[0079] Table 6 EPMT activity status of mice subjected to chronic unpredictable mild stress, n=12

[0080]

[0081] Note: Compared with the normal group, #p<0.05, ##p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0082] The results showed that compared with the normal group animals, the model group animals entered the open arms significantly less times and stayed in the open arms less time in the elevated plus maze. After treatment with different components of Huoshan Dendrobium extract, the number of times mice entered the open arms in the elevated plus maze increased significantly, and the time they stayed in the open arms increased significantly, among which the water layer component of Huoshan Dendrobium extract had the best effect. This shows that Huoshan Dendrobium extract can increase the amount of exercise and central area exploration ability of depression model mice in the open field, and improve the overall activity state of depression mice.

[0083] Table 7 Changes of inflammatory factors in mice subjected to chronic unpredictable mild stress, n=12

[0084] Group TNF-α(ng / L) IL-1β (pg / mL) IL-6 (pg / mL) BDNF (pg / mL) Control 152.43±33.14 80.33±8.53 46.15±3.48 524.64±45.17 Model <![CDATA[544.35±62.78 ## ]]> <![CDATA[242.06±25.90 ## ]]> <![CDATA[172.31±18.45 ## ]]> <![CDATA[115.45±31.44 ## ]]> FLU <![CDATA[192.60±48.15 ** ]]> <![CDATA[106.78±18.22 ** ]]> <![CDATA[67.08±7.18 ** ]]> <![CDATA[476.12±78.42 ** ]]> DHPE <![CDATA[230.55±56.32 ** ]]> <![CDATA[213.24±24.88 * ]]> <![CDATA[138.96±13.45 ** ]]> <![CDATA[257.42±46.33 ** ]]> DHEA <![CDATA[286.42±49.42 ** ]]> <![CDATA[180.44±14.78 ** ]]> <![CDATA[108.14±10.24 ** ]]> <![CDATA[288.44±53.17 ** ]]> HkDJ <![CDATA[332.18±59.70 ** ]]> <![CDATA[142.11±16.47 ** ]]> <![CDATA[95.66±8.63 ** ]]> <![CDATA[312.35±69.05 ** ]]> DHW <![CDATA[190.27±27.45 ** ]]> <![CDATA[90.34±8.55 ** ]]> <![CDATA[54.68±6.45 ** ]]> <![CDATA[494.24±75.78 ** ]]>

[0085] The results showed that compared with the normal group, the pro-inflammatory factors TNF-α, IL-1β and IL-6 in the model group increased significantly, and BDNF decreased significantly. After treatment with different components of Huoshan Dendrobium extract, the release of inflammatory factors TNF-α, IL-1β and IL-6 was significantly inhibited, and the synthesis of BDNF was promoted, among which the water layer component of Huoshan Dendrobium extract had the best effect. This shows that Huoshan Dendrobium extract can improve the inflammatory level of depression model mice and improve the overall physiological state of depression mice.

[0086] Conclusion: The extract of Dendrobium huoshanense has significant antidepressant effect, has good efficacy in preventing and / or treating depression, and has good application prospects in the prevention and / or treatment of depression.

[0087] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A Dendrobium huoshanense extract, characterized in that The Huoshan Dendrobium extract is obtained by reflux extraction using an alcohol-water mixture, wherein the volume concentration of alcohol in the alcohol-water mixture is 50%-90%.

2. The Dendrobium huoshanense extract according to claim 1, characterized in that The volume concentration of alcohol in the alcohol-water mixture is 70%.

3. The Dendrobium huoshanense extract according to claim 1, characterized in that The conditions of the reflux extraction include: temperature of 70-90° C. and time of 1-4 h.

4. The Dendrobium huoshanense extract according to claim 3, characterized in that The temperature is 85° C. and the time is 2 hours.

5. The Dendrobium huoshanense extract according to claim 1, characterized in that The preparation method of the Huoshan Dendrobium officinale extract comprises: firstly subjecting the stems of the Huoshan Dendrobium officinale to reflux extraction with a mixture of alcohol and water, and then subjecting the stems to decompression and concentration until there is no alcohol taste to obtain a suspended aqueous solution; extracting the mixture with petroleum ether, ethyl acetate and n-butanol in sequence, and finally obtaining solutions of each component, and subjecting the solutions to decompression, concentration and drying to obtain the Huoshan Dendrobium officinale extract.

6. Use of the Dendrobium huoshanense extract according to any one of claims 1 to 5 in the preparation of a medicament for preventing and / or treating depression.

7. The use according to claim 6, characterized in that: The depression is caused by chronic unpredictable mild stress.

8. The use according to claim 6, characterized in that: The depression includes at least one of depression accompanied by anxiety and depression accompanied by mania.

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