CN111494406A - Application of Ganoderma lucidum water decoction and its polysaccharides in the preparation of anti-anxiety drugs - Google Patents

Abstract

An application of ganoderma lucidum water decoction and polysaccharide thereof in preparing anxiolytic drugs is disclosed, wherein the ganoderma lucidum water decoction comprises the following components: pulverizing Ganoderma decoction pieces, reflux-extracting with distilled water for 3 times (each for 1 hr), mixing the three filtrates, and concentrating under reduced pressure; the ganoderma lucidum polysaccharide is: pulverizing Ganoderma decoction pieces, extracting with ethanol under reflux for 3 times, defatting, and oven drying; adding distilled water into dried ganoderma lucidum decoction pieces, carrying out reflux extraction for 3 times, each time for 2 hours, carrying out suction filtration to obtain supernatant, carrying out reduced pressure concentration on the supernatant, placing the supernatant to room temperature, adding absolute ethyl alcohol, standing for 24 hours at low temperature, centrifuging, collecting precipitate, volatilizing until no alcohol smell exists, dissolving the precipitate in distilled water, centrifuging to remove insoluble substances to obtain supernatant, adding absolute ethyl alcohol into the supernatant, standing, centrifuging, collecting precipitate, and freeze-drying.

Description

Application of Ganoderma lucidum water decoction and its polysaccharides in the preparation of anti-anxiety drugs

technical field

The invention relates to the field of medicine, in particular to the application of a ganoderma lucidum water decoction and its polysaccharide in the preparation of anxiolytic drugs.

Background technique

Ganoderma lucidum, the dried fruiting body of the Polypore family fungus Ganoderma lucidum (Leyss.exFr.) Karst.

"The Yellow Emperor's Classic of Internal Medicine" records: "Ganoderma lucidum enters the five meridians, nourishes the Qi of the five internal organs, regulates the yin and yang of the six internal organs, keeps the righteousness in the body, and the evil cannot dry"; Replenishing the middle, increasing wisdom and not forgetting. Eating for a long time will not age and prolong life”; “Xinxiu Materia Medica” records “Ganoderma lucidum is at ease.” It is calm in nature, sweet in taste, and returns to the heart, lung, liver and kidney meridians.

The active ingredients of Ganoderma lucidum are complex. Among them, polysaccharides, triterpenoids and proteins are the main active ingredients of Ganoderma lucidum. Among them, Ganoderma lucidum polysaccharide has the highest content and is its key medicinal ingredient. Modern pharmacological studies have shown that Ganoderma lucidum has anti-tumor, antioxidant, immune regulation, hypoglycemic, sedative and hypnotic, and hepatoprotective effects. It is clinically used for chronic bronchitis and repeated respiratory infections, hypertension, diabetes, neurasthenia, insomnia, Treatment of diseases such as viral hepatitis. However, there is no relevant research report on the anti-anxiety effect of Ganoderma lucidum.

SUMMARY OF THE INVENTION

In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide an application of Ganoderma lucidum water decoction and its polysaccharide in the preparation of anti-anxiety drugs, which can effectively solve the problem of preparing anti-anxiety drugs.

The technical scheme solved by the present invention is the application of a Ganoderma lucidum water decoction and its polysaccharides in the preparation of anti-anxiety drugs. Extract for 1 hour, combine the filtrates three times, and concentrate under reduced pressure to a concentration equivalent to a ganoderma lucidum water decoction containing crude drug 0.2g·mL ^-1 -0.6g·mL ^-1 ; the ganoderma polysaccharide is: pulverize ganoderma lucidum pieces, add a volume of The concentration is 95% ethanol reflux extraction 3 times, degreasing, 2h each time, drying the degreasing ganoderma lucidum pieces; adding the dried ganoderma lucidum pieces into distilled water for reflux extraction 3 times, 2h each time, combining the three filtrates, suction filtration, to obtain The supernatant was concentrated under reduced pressure, placed at room temperature, then added with absolute ethanol until the alcohol content reached 80% by volume, stood at low temperature for 24 hours, centrifuged to collect the precipitate, evaporated until there was no alcohol smell, and the precipitate was dissolved in distilled water , centrifuge to remove insolubles to obtain a supernatant, add absolute ethanol to the supernatant until the alcohol content reaches 80% by volume, stand for centrifugation, collect the precipitate, and freeze-dry to obtain Ganoderma lucidum polysaccharide powder (crude sugar).

The Ganoderma lucidum water decoction and Ganoderma lucidum polysaccharide prepared by the above-mentioned Ganoderma lucidum have anti-anxiety effects, are effectively used for the preparation of anti-anxiety drugs, and realize the application in the preparation of anti-anxiety drugs.

The present invention has rich raw materials, simple preparation method, and the product has anti-anxiety effect, opens up new medicinal value of Ganoderma lucidum and a new way of anti-anxiety medicine, is an innovation in medicine for treating anxiety disorder, and has significant social and economic benefits.

Detailed ways

The specific embodiments of the present invention will be described in detail below with reference to specific conditions and examples.

Example 1

An application of Ganoderma lucidum water decoction in the preparation of anti-anxiety drugs, the Ganoderma lucidum water decoction is: taking ganoderma lucidum pieces and pulverizing, adding distilled water for reflux extraction 3 times, adding 10 times the volume of distilled water each time, the weight of the The volume refers to the solid in g and the liquid in ml (the same below). During the first reflux, soak for 30 minutes, extract for 1 hour each time, combine the three filtrates, and concentrate under reduced pressure to a concentration equivalent to 0.2 g of crude drug. mL ^-1 -0.6g·mL ^-1 of Ganoderma lucidum decoction;

The above-mentioned Ganoderma lucidum water decoction has an anti-anxiety effect, is effectively used for the preparation of anxiolytic drugs, and realizes the application in the preparation of anti-anxiety drugs.

Example 2

An application of Ganoderma lucidum polysaccharide in the preparation of anxiolytic drugs, the Ganoderma lucidum polysaccharide is: crushing ganoderma lucidum pieces, adding 10 times the weight and volume of ethanol with a volume concentration of 95% for reflux extraction 3 times, defatting, 2h each time, During the first reflux, soak for 30 minutes, collect the degreasing ganoderma lucidum pieces for three times, and dry them at 40 °C; add the dried ganoderma lucidum pieces into 12 weight volumes of distilled water for reflux extraction for 3 times, 2 hours each time, during the first reflux, soak 30min, combine the filtrates three times, and filter them with suction to obtain the supernatant. The supernatant is placed in a rotary evaporator, concentrated under reduced pressure, placed at room temperature, and anhydrous ethanol is slowly added while stirring until the alcohol content reaches 80% by volume. , stand at 4°C for 24h, centrifuge at 3500r/min for 15min, collect the precipitate, volatilize until there is no alcohol smell, add distilled water to dissolve, centrifuge to remove insoluble matter, and obtain a supernatant. The alcohol content reaches 80% by volume, stand at 4°C for 24h, centrifuge at 3500r/min for 15min, collect the precipitate, freeze-dry to obtain Ganoderma lucidum polysaccharide powder (crude sugar), and the yield of Ganoderma lucidum polysaccharide powder is about 1.8%.

The Ganoderma lucidum polysaccharide prepared from the above-mentioned Ganoderma lucidum has anxiolytic effects, is effectively used for the preparation of anxiolytic drugs, and realizes the application in the preparation of anti-anxiety drugs.

The invention has rich raw materials, simple preparation method and easy production, and the obtained product has anti-anxiety effect, especially can increase the content of GABA in the brain, reduce the content of GLU, 5-HT and DA, and is effectively used for preparing anxiolytic drugs, and After experiments, very good technical results have been achieved, and the relevant experimental data are as follows.

1. Effect of Ganoderma lucidum decoction on mouse anxiety model.

1 Experimental material

1.1 Experimental animals

SPF grade healthy male Kunming mice, weighing 18-22 g, were purchased from Henan Provincial Laboratory Animal Center, license number: SCXK (Yu) 2017-0001, batch number: NO.DW2019060015. The mice were kept in the IVC-II mouse independent air supply isolation cage of the Animal Experiment Center of Henan University of Traditional Chinese Medicine, and the laboratory temperature was controlled at 22 ± 2 °C.

1.2 Experimental drugs

Ganoderma lucidum is the dried fruiting body of the Polyporaceae fungus Ganoderma lucidum (Leyss.exFr.) Karst. The place of origin, Liaoning, was purchased from Henan Qianfang Pharmaceutical Co., Ltd., batch number: 180601QF. It has been identified as genuine Ganoderma lucidum by the School of Pharmacy, Henan University of Traditional Chinese Medicine.

Diazepam, purchased from the Third Affiliated Hospital of Henan University of Traditional Chinese Medicine, manufacturer: Tianjin Lisheng Pharmaceutical Co., Ltd., specification: 2.5mg/tablet × 100 tablets, approval number: Guoyao Zhunzi H12020119, production batch number: 1806008.

Gamma-aminobutyric acid (GABA) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Glutamic acid (GLU) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Serotonin (5-HT) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Dopamine (DA) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.

1.3 Experimental instruments

Rat IVC Independent Air Supply Isolation Cage (Model: IVC-II, Suzhou Fengshi Laboratory Animal Equipment Co., Ltd.);

PM-200 Elevated Cross Labyrinth Device: Chengdu Taimeng Technology Co., Ltd.;

The mouse light-dark box device was purchased from Shanghai Xinsoft Information Technology Co., Ltd.;

Microplate reader (model: Epoch) was purchased from BioTeK Instruments, Inc.

2 Experimental methods

2.1 Experimental drugs

2.1.1 Ganoderma lucidum water decoction prepared by the present invention, the Ganoderma lucidum water decoction is made to be equivalent to containing crude drug 0.2g·mL ^-1 (low content), 0.4g·mL ^-1 (medium content) and 0.6g·mL respectively. ^-1 (high content) of Ganoderma lucidum decoction.

2.1.2 Preparation of Diazepam Suspension: Take diazepam tablets and grind them, dissolve in distilled water, and sonicate to prepare a diazepam suspension of 0.25 mg·mL ^-1 for use. Shake well before use.

2.2 Grouping and Administration

72 healthy male Kunming mice of SPF grade, weighing 18-22 g, were adaptively reared for 3 days, labeled with picric acid and weighed, and randomly divided into 6 groups, namely blank control group (without behavioral experiment), anxiety Model group, positive control group, Ganoderma lucidum decoction high, medium and low dose groups, 12 in each group. Mice were screened and grouped by an open-box experiment 1 day before administration to ensure that there was no significant difference in the behavioral abilities of the mice in each group. Oral administration was administered every morning for 5 consecutive days, and the administration volume was 0.1 mL·10 g ^-1 . The blank control group and anxiety model group were intragastrically administered with distilled water every day, the positive control group was intragastrically administered 0.0025 g·kg ^-1 diazepam suspension each day, and the high, medium and low dose groups of Ganoderma lucidum decoction were intragastrically administered 6 g·kg kg ^-1 , 4g·kg ^-1 , 2g·kg ^-1 of Ganoderma lucidum water decoction, the behavioral test was performed 1h after the last intragastric administration.

2.3 Behavioral tests

2.3.1 Elevated plus maze experiment: Before the experiment, each mouse was placed in a separate cage for free movement for 5 minutes, and then the mice were placed on the open platform in the center of the elevated plus maze, with the head facing the open arm. Let it explore freely. The computer performed tracking recording to collect the movement data of the mice on the elevated plus maze, and the recording time was 5 min. Taking OE%, that is, the number of times of opening the arm of the mouse/(the number of entering the open arm + the number of entering the closed arm), and OT%, that is, the open arm retention time/(open arm retention time + closed arm retention time) as the evaluation of Ganoderma lucidum water decoction. An indicator of the anti-anxiety effect of liquid. After the test of each mouse, the excrement such as feces should be removed in time, and if necessary, the maze should be sprayed with low-concentration alcohol and wiped to remove the odor, and then the next test will be carried out. During the whole experiment process, it is necessary to keep the experimental environment quiet and the experimental equipment clean, and try to avoid the adverse effects of the external environment on the experimental results. Note: Each mouse can only be used once. If the device is dropped during the experiment, the experiment for this mouse is over and the experiment cannot be repeated.

2.3.2 Light and dark box experiment: After the elevated plus maze experiment, the light and dark box experiment should be carried out immediately. First, let the mice move freely in a quiet environment for 5 minutes, and then put the mice in the center of the light box with their backs facing the dark box. The shuttle times of mice between the light box and the dark box were observed, and the recording time was 10 minutes, which was used as an index to evaluate the anxiolytic effect of Ganoderma lucidum decoction. After the test of each mouse, remove excrement such as feces in time, spray and wipe the bottom of the box with low-concentration alcohol if necessary to remove odor, and then proceed to the next test. During the whole experiment process, it is necessary to keep the experimental environment quiet and the experimental equipment clean, and try to avoid the adverse effects of the external environment on the experimental results.

2.4 Detection of relevant indicators

Immediately after the behavioral experiment, the mice were sacrificed by cervical dislocation, the head was quickly cut off, and the whole brain was quickly peeled off on the ice table, wrapped in tin foil, with the corresponding number written, and quickly put into liquid nitrogen for storage, and then transferred. Store in a refrigerator at -80°C for the determination of GABA, GLU, 5-HT and DA. Accurately weigh with a 1/10,000 balance, add ice-cold saline at a ratio of brain tissue (g): saline (mL) = 1:4, and use a grinder and a vortex shaker to pulverize and homogenize at low temperature. , the homogenate was centrifuged at 4°C, 3500r·min ^-1 for 20min, then the supernatant was aspirated and placed in a 1.5mL centrifuge tube to obtain a mouse brain tissue homogenate with a concentration of 20%. Store in a -80°C refrigerator. The contents of GABA, GLU, 5-HT and DA in mouse brain tissue were determined by enzyme-linked immunosorbent assay in strict accordance with the kit instructions.

2.5 Statistical processing

)表示，以P<0.05表示有显著性差异，P<0.01表示有极显著性差异。The experimental data were analyzed by one-way analysis of variance using SPSS 23.0 software, and the data in each group were expressed as mean ± standard deviation (

) indicates that there is a significant difference with P<0.05, and a very significant difference with P<0.01.

3 results

3.1 Effect of Ganoderma lucidum decoction on behavioral experiments of mouse anxiety model

Compared with the anxiety model group, diazepam in the positive control group significantly increased the number of shuttles in the light-dark box test (P<0.05), and significantly increased the OE% (P<0.01) and OT of the mice in the elevated plus maze test. % (P<0.01), indicating that the modeling was successful; compared with the anxiety model group, the OE% of the mice in the Ganoderma lucidum water decoction group was significantly increased in the elevated plus maze test (P<0.05); Ganoderma lucidum water decoction groups in each dose group The OT% of mice in the elevated plus maze test showed an increasing trend, but there was no significant difference compared with the anxiety model group; the mice in the middle-dose Ganoderma lucidum decoction group had a significant increase in the number of shuttles in the light-dark box test (P<0.05) . This indicates that Ganoderma lucidum decoction has a certain intervention effect on the anxiety behavior of anxiety model mice. See Table 1.

n＝12)Table 1 Effects of Ganoderma lucidum decoction on the behavior of elevated plus maze and light-dark box in mice (

n=12)

Note: Compared with the anxiety model group, #P<0.05, ##P<0.01.

3.2 The effect of Ganoderma lucidum decoction on the content of GABA, GLU, 5-HT and DA in the brain tissue of mouse anxiety model

Compared with the blank control group, the content of GABA in the brain tissue of the anxiety model group was significantly decreased (P<0.01), and the content of GLU was not significantly different, but the ratio of GLU to GABA was significantly increased (P<0.05), which was significantly different from the anxiety model group. Compared with other groups, the content of GABA in the brain tissue of the mice in the positive control group and the high-dose Ganoderma lucidum decoction group was significantly increased (P<0.05, P<0.01); The content of GLU in the mice was significantly decreased (P<0.05, P<0.01); the ratio of GLU to GABA in the brain tissue of the mice in the positive control group and the high-, medium- and low-dose Ganoderma lucidum decoction groups was significantly decreased (P<0.05, P<0.01). 0.01). See Table 2.

Compared with the blank control group, the content of 5-HT in the brain tissue of the mice in the anxiety model group had a trend of increasing, and the content of DA had a trend of decreasing, but it was not statistically significant. Compared with the anxiety model group, the water decoction of Ganoderma lucidum was higher The content of 5-HT in the brain tissue of mice in the , middle and low dose groups was significantly decreased (P<0.01), and the positive control group had a significant decreasing trend; The content decreased significantly (P<0.05, P<0.01), and the positive control group had an increasing trend. See Table 3.

n＝12)Table 2 Effect of Ganoderma lucidum decoction on GABA and GLU content in the brain tissue of mouse anxiety model (

n=12)

Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the anxiety model group, ^# P<0.05, ^## P<0.01.

n＝12)The effect of table 3 Ganoderma lucidum water decoction on the content of 5-HT and DA in the brain tissue of mouse anxiety model (

n=12)

Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the anxiety model group, ^# P<0.05, ^## P<0.01.

2. Effect of Ganoderma lucidum polysaccharide on mouse anxiety model

1 Experimental material

1.1 Experimental animals

SPF grade healthy male Kunming mice, weighing 18-22 g, were purchased from Henan Provincial Laboratory Animal Center, license number: SCXK (Yu) 2017-0001, batch number: NO.DW2019080046. The mice were kept in the IVC-II mouse independent air supply isolation cage of the Animal Experiment Center of Henan University of Traditional Chinese Medicine, and the laboratory temperature was controlled at 22 ± 2 °C.

1.2 Experimental drugs

Ganoderma lucidum is the dried fruiting body of the Polyporaceae fungus Ganoderma lucidum (Leyss.exFr.) Karst. The place of origin, Liaoning, was purchased from Henan Qianfang Pharmaceutical Co., Ltd., batch number: 180601QF. It has been identified as genuine Ganoderma lucidum by the School of Pharmacy, Henan University of Traditional Chinese Medicine.

Diazepam, purchased from the Third Affiliated Hospital of Henan University of Traditional Chinese Medicine, manufacturer: Tianjin Lisheng Pharmaceutical Co., Ltd., specification: 2.5mg/tablet × 100 tablets, approval number: Guoyao Zhunzi H12020119, production batch number: 1806008.

Gamma-aminobutyric acid (GABA) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Glutamic acid (GLU) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Serotonin (5-HT) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.;

Dopamine (DA) kit (batch number E20190801A) was purchased from Shanghai Alessa Biotechnology Co., Ltd.

1.3 Experimental instruments

Rat IVC Independent Air Supply Isolation Cage (Model: IVC-II, Suzhou Fengshi Laboratory Animal Equipment Co., Ltd.);

PM-200 Elevated Cross Labyrinth Device: Chengdu Taimeng Technology Co., Ltd.;

The mouse light-dark box device was purchased from Shanghai Xinsoft Information Technology Co., Ltd.;

Microplate reader (model: Epoch) was purchased from BioTeK Instruments, Inc.

2 Experimental methods

2.1 Preparation of drugs

2.1.1 Ganoderma lucidum polysaccharide (crude polysaccharide) prepared by the present invention, Ganoderma lucidum polysaccharide is prepared into Ganoderma lucidum crude polysaccharide aqueous solution respectively, and the concentration is respectively 12mg·mL ^-1 (high concentration), 8mg·mL ^-1 (medium concentration), 4mg·mL mL ^-1 (low concentration).

2.1.2 Preparation of Diazepam Suspension: Take diazepam tablets and grind them, dissolve in distilled water, and sonicate to prepare a diazepam suspension of 0.25 mg·mL ^-1 for use. Shake well before use.

2.2 Grouping and Administration

Seventy-two SPF-grade healthy male Kunming mice, weighing 18-22 g, were labeled with picric acid and weighed after adaptive feeding for 3 days. They were randomly divided into 6 groups, namely blank control group (without behavioral experiment), anxiety model group, positive control group, high, medium and low dose groups of Ganoderma lucidum crude polysaccharide, 12 in each group. Mice were screened and grouped by an open-box experiment 1 day before administration to ensure that there was no significant difference in the behavioral abilities of the mice in each group. Oral administration was administered every morning for 5 consecutive days, and the administration volume was 0.1 mL·10 g ^-1 . The blank control group and the anxiety model group were given distilled water by gavage every day, the positive control group was given 0.0025 g·kg ^-1 diazepam suspension by gavage every day, and the Ganoderma lucidum polysaccharide high, medium and low dose groups were given 120 mg·kg by gavage, respectively. ^-1 , 80 mg·kg ^-1 , 40 mg·kg ^-1 aqueous solution of Ganoderma lucidum crude polysaccharide. Behavioral tests were performed 1 hour after the last intragastric administration.

2.3 Behavioral tests

The elevated plus maze experiment and the light-dark box experiment are the same as the methods under "2.3 Behavioral Test" in the previous section.

2.4 Detection of relevant indicators

Detect with the same method under "2.4 Detection of Related Indicators" in the previous part.

2.5 Statistical processing

表示，以P<0.05表示有显著性差异，P<0.01表示有极显著性差异。The experimental data were analyzed by one-way analysis of variance using SPSS 23.0 software, and the data in each group were expressed as mean ± standard deviation.

Indicates that there is a significant difference with P<0.05, and a very significant difference if P<0.01.

3 results

3.1 Effects of Ganoderma lucidum crude polysaccharide on behavioral experiments of mouse anxiety model

Compared with the anxiety model group, diazepam in the positive control group could significantly increase the OE% (P<0.01), OT% (P<0.01) of the mice in the elevated plus maze test and the number of shuttles in the light-dark box test (P<0.01). Compared with the anxiety model group, the OE% of the mice in the low-dose Ganoderma lucidum polysaccharide group increased significantly in the elevated plus maze test (P<0.05), and the mice in the Ganoderma lucidum high-dose and medium-dose groups The OE% of Ganoderma lucidum polysaccharide group showed a significant increase trend, but there was no significant difference compared with the anxiety model group; the OT% of the mice in the high, medium and low dose groups of Ganoderma lucidum polysaccharide were significantly increased in the elevated plus maze test (P<0.05, P<0.05, P<0.05). 0.01); the shuttle times of Ganoderma lucidum polysaccharide medium dose group increased significantly (P<0.05), and the shuttle times of Ganoderma lucidum crude polysaccharide high and low dose groups increased significantly, but compared with anxiety model group No significant difference. It shows that Ganoderma lucidum polysaccharide has a good intervention effect on the anxiety behavior of anxiety model mice. See Table 4.

n＝12)Table 4 Effects of Ganoderma lucidum polysaccharides on the behavior of elevated plus maze and light-dark box in mice (

n=12)

Note: Compared with the anxiety model group, ^# P<0.05, ^## P<0.01.

3.2 Effects of Ganoderma lucidum crude polysaccharide on the content of GABA, GLU, 5-HT and DA in the brain tissue of mouse anxiety model

Compared with the blank control group, the content of GABA in the brain tissue of the anxiety model group was significantly decreased (P<0.01), the content of GLU was significantly increased (P<0.01), and the ratio of GLU to GABA was significantly increased (P<0.01). , compared with the anxiety model group, the GABA content in the brain tissue of the mice in the positive control group and the low-dose Ganoderma lucidum polysaccharide group was significantly increased (P<0.05, P<0.01); The content of GLU in the brain tissue of mice was significantly decreased (P<0.05, P<0.01); the ratio of GLU to GABA in the brain tissue of the mice in the positive control group and the high, medium and low dose groups of Ganoderma lucidum polysaccharide was significantly decreased (P<0.05). , P<0.01). See Table 5.

Compared with the blank control group, the content of DA in the brain tissue of the anxiety model group decreased significantly (P<0.01), and the content of 5-HT increased significantly, but there was no significant difference compared with the blank control group. Compared with the anxiety model group, the content of 5-HT in the brain tissue of the Ganoderma lucidum crude polysaccharide medium and low dose groups was significantly decreased (P<0.05, P<0.01), and the positive control group had a significant decreasing trend; the Ganoderma lucidum crude polysaccharide medium dose group The content of DA in mouse brain tissue was significantly decreased (P<0.05). See Table 6.

n＝12)Table 5 Effect of Ganoderma lucidum polysaccharide on GABA and GLU content in the brain tissue of mouse anxiety model (

n=12)

Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the anxiety model group, ^# P<0.05, ^## P<0.01.

n＝12)Table 6 Effect of Ganoderma lucidum crude polysaccharide on the content of 5-HT and DA in the brain tissue of mouse anxiety model (

n=12)

Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the anxiety model group, ^# P<0.05, ^## P<0.01.

4 Conclusion

The results showed that the water decoction of Ganoderma lucidum and its crude polysaccharides showed good anxiolytic effects on the anxiety model in mice. DA content.

The above clearly shows that the present invention uses the elevated plus maze experiment and the light-dark box shuttle experiment to conduct anti-anxiety research, and uses the high-efficiency Elisa method to measure the contents of GABA, GLU, 5-HT, and DA in the brain of mice. The percentage of open-arm retention time and the percentage of open-arm entry times, and the number of shuttles in the light and dark boxes were used to test the anxiolytic effects of Ganoderma lucidum water decoction and Ganoderma lucidum crude polysaccharide; by comparing GABA, GLU, 5- HT and DA content to explore the anti-anxiety mechanism of Ganoderma lucidum and its crude polysaccharides, which provides technical support for the development of new medicinal and commercial values of Ganoderma lucidum. It is a major innovation in anti-anxiety drugs and has significant economic and social benefits.

Claims (3)

1. An application of a water decoction of Ganoderma in preparing anxiolytic drug is characterized in that the water decoction of Ganoderma is prepared by pulverizing decoction pieces of Ganoderma, adding distilled water, reflux-extracting for 3 times, each time adding 10 times of distilled water, the weight volume is solid in g, liquid in ml, soaking for 30min during first reflux, reflux-extracting for 1h, mixing filtrates, concentrating under reduced pressure to concentration equivalent to 0.2 g.m L of crude drug^-1-0.6g·mL^-1The water decoction of the lucid ganoderma;

the application of ganoderma lucidum polysaccharide in preparing anxiolytic drugs is characterized in that the ganoderma lucidum polysaccharide is: pulverizing Ganoderma decoction pieces, adding 10 times of ethanol with volume concentration of 95% and reflux extracting for 3 times, defatting for 2 hr, soaking for 30min during first reflux, collecting Ganoderma fragments after three defatting, and oven drying at 40 deg.C; adding distilled water with the volume of 12 weight into the dried ganoderma lucidum fragments, carrying out reflux extraction for 3 times, each time for 2 hours, soaking for 30 minutes during first reflux, combining three filtrates, carrying out suction filtration to obtain a supernatant, placing the supernatant in a rotary evaporator, carrying out reduced pressure concentration, placing to room temperature, slowly adding absolute ethyl alcohol while stirring until the alcohol content reaches 80% by volume, standing for 24 hours at 4 ℃, centrifuging for 15 minutes at 3500r/min, collecting precipitates, volatilizing until no alcohol smell exists, adding distilled water for dissolving, centrifuging to remove insoluble substances to obtain a supernatant, slowly adding absolute ethyl alcohol while stirring again until the alcohol content reaches 80% by volume, standing for 24 hours at 4 ℃, centrifuging for 15 minutes at 3500r/min, collecting precipitates, and carrying out freeze drying to obtain the ganoderma lucidum polysaccharide.

2. The application of the ganoderma lucidum water decoction of claim 1 in preparing an anxiolytic medicament, which is characterized by being used for preparing a medicament for improving the symptoms of anxiety, increasing the GABA content in brain and reducing the G L U content, 5-HT content and DA content.

3. The use of the ganoderan according to claim 2 for the preparation of anxiolytic drugs, characterized by the use for the preparation of drugs that improve the symptoms of anxiety and increase the GABA content in the brain, decreasing the G L U and the 5-HT and DA content.